I
. 
4r. .- 

..J. 

h-

.

  - cu'mms ABD mnwmimzipmum. a; . 
52.1  memoirs :3 ma 2,h.5 ACID magma .

 
    
  
 
   
     
    
 
 
  
   
   
     
  
   
    
   
   
  
  
  
  
   

 

amalgam 31! 1:9: Grimm. warrant, sumo?It; . .IItimeoutsdotm'aina the "palatial landmiuity --

man

I
1..
-

55421914 M00 9

. 

  cl  - .  -.



       
  

 

 ?w'irl v- . - -, r. -ulsic-f I  1.163;:
mi   mule     I ..

 gigs-=13 mama thro? (3) #Mm mom  

    

   

     

            

mall  :21th- _tmiur??rmnlad {Jam mam-1W2. 
-. . . --    .7 
:evlith' mamm.m par-fumed an mbhits.? lane 01: discussed}.  
I, 51f?li$10th the "thank amTau. l'  - "f I
 m2mama to he the #151?: mu -r

 

 

   

 

  

-

    

a. .11? 

 

[m mama mudiw 
{pm a: "epithelial mamas; mom"    

?r .1
- 

?1

sf?gmtimphamoc, and finally, pawn? mm'tf

 ?m mm mm. mm mm ?8146

this

.I..



 me? ?am? in ma?mmluv;
1



   

 

  
     

 

-.-

- - a  945)?. 
- - - - 
a .- n, . 

   
  

 
   

I

   

   
  
 
    
  
  
 
 
 
 
 
 
  
    

-- -2-

'i?ha obgarmtiona of Parnell ?on tho postnatal amlopamt of tha 

GHISL M00-

"bsoaqna glands m? the, putth amg? ta mung: mm or. animating

3159msih??rxmnof  to
.lfr'; 5.3-. 1 .

 

.iv.
4-  


a. .

. 

 13.55  27.   ?nll  and ?lls   7  
.  with at ?ames of (hascalla  in 1.136 __matnry



i  magi. rim'

 can: anti-PM? ?19151? .  
- mm 

"?xy'v 

  
  

 
   
 
   
 


. -. at
c.

     
  

     
    
   

   
   

   
   

Eras-5a: . - 

 
   
  

2m m: a. ?ram-11:: mow-311m- phan? 91" ?ue pal-harps min gluing-a 
 .V .: - ?If: .: -.. . -- 15? f-l-u -.--.  i .
 .- . - an! ducts mum-Joins: in m. -:

gnaw-means cuppa-tad meander tho rut animal. Firatlm hair m1. and concomitant. 191ml dung,rat we well know; 3 Sammyrmmim'bln h?tohm. than available data on the errant of upplying mien: ?bula}. mince; 
. . - .   

10 the quid-mun -..-.-.   -- 

. ., s. . A.  I,   
 War-.144:14,-
I . '17 I. 

 

vain?.4: -

 
    
   
    
  
    
  

.
u'qg(in?considaratiun of mung paging-M gland -. 




 . .. . ti; ME #1591?  it ?i?swim: A that.  


 

 mum is; m" maze?  grim??  W357 .320.  
uni-mew; 2: i. Imam mW??l?fjhEW.

 


-. .
- -..- 

1-.

   
     
 

    

             

 -. - ..  .-    :1  
  innjba?avnlogiag'satnoeoul   
I
 .?iwz: .   -. 
.nli?fgf??i?}:  ~39?  m.   WPH  7.   

.  - 54.;    -  - -


- 113a (?ring mam may 



      01' ?lm pro?ts 



  hm #gnagm: to the skin  31m  

alv-

. 
.6 . 

 rem-am 
I I  any Wait-nation fem-ding  taxi.an 9f." - 16 
  MW. Swiss! or ..

  

6d.  :3 :81;  - #:6qu  

a . r?  ?1341."   "fir-5?  



1049.Z-n'f 

-. -
5:5. .-
- 
.

- rt.- - .I:

      
  
 

 

 

 
    
 

.-- ?nu-.1 "Ltlv;'fl "57"

Inn-7:, - Ill 
 

. --.

HI ?rm. :r?r

  
   
  

  

  
  
  

   
  
  

 
 
 
 

 mammal Animist

nude

-

 mu  in thin Laboratory tam: in  ob." 

TSLISL

 

-

-    ,tninad  Caz-warm Flu-an, Backhand,  fork. . 
I  (-HMS:  

- 
?n


'm   I.  

  
      

swung. auxin  
insignia.th    

           
  

- -
 


.

. 3.11:?am?2,1; I - . 

V?s-r -. 1? . .-.. . -- ?Maxim? 

 

ta'at?mqmumm uhed for' ayplinatinn?to the. am ruptured  

?ght except 113' an  o?f 

 

Oaks? mm: diaaolvad 1n sat-er.

?u tun-?4?; 





 bmz-imnga  

4-  

   
    
  
 

A def-n taut-litaI-m-n'ing mm??mn. 18149

?uaud mm at: minim-1:15. It 

   

-- - I I I?d? -. .
?dgui; a;  

 
- ot-J?hgmma 

-- 31-9.

    
   
 

.. - ?if  
.

 

as-

Mawhas. 2 in length during-tho 

. w: um.? -  Mudml's 

- 

 ?ema- m?    

33924191thug _upmnonts, tin (S) hilly trotted applications

 dado inch day, The    

  

 -.    am and: whim appliad ?t'ad Ind ream-M  

   .  151' km??th 

    
  
  
   
  
 
     
   
   
 

. 

. Ecltw 



may. (we Mk: nu-  iwm. in   
1? '1 I 

In

1.513

W7 . 3m garawf "f  .  j'    
3'14: - 





- 
- . ..-. - -. --
. .with  - 

  

  
    

    
    

 
   
 



 
  

  
 

-

 

 
 

- .. ritamv  "aim" .13: .
. . Fl ?Navy?? j. - - -

- ;?Anp'Wham period: or varying?dnnu More

. - - 1.?au.aphases  hair? I.  5 

   Wham ma coma by treating A hue;-

was inn much uaad a for th?  bung
??lf 5.41.5:  I apps-'ni'aed. other controls were used in (tuba-Int phhbou at"

tho total Wm process, Ind ?warn.?    
 gim?gpregi 


- syn-:- -, 1:1?  'dr?aar .
. -.. .fzxzi  ?g..me ?italinil? - .i 

   . . I. a. amend; tho skin or the M1 was malted m- with
  -  I a dry camel's hair brush for 15 times.

ngusa MOO

a, run-9mm: tho

9f ?aw-1 in!  'i-i-irand located wax-the matu- or the 3mm was   

dun "Am painted any with can of ua'maral

:nmm may for marine}  or was as mu-
   . -ntad 1n Tablas Iv and v. can sit- unsaved at 
I a mw, and mmg'. I
   

  
  





 
 
  
  
      

 
 

 Mama and gained rims  ma." 

ifan:  m-.32941:?? +35I.-

J.'tun-3m" ga?tgm??rg-I; "at-tax. - 3-, I  .3321--.- -- 15-.- . n' 9-4-4: - w;  ?ab 3- 

D. armory of  Pmoedm'oa;   

   WIS or ?an Miami: an mart:in in Tablesfollows(Tabla I, Sections A and B). . 
u,

I 2,h,5 and u. control mbs?mm war. applied 
mucus to the skin of Wan; rats. Thane Applicatin    

. 12.1.mI-Iu

  


-.--

gs 



mug; Moo



   viewable and the control abut-men! m0 @1311?

 

i   I to skin of mm 1'3? ?m8 W115 Pm?? ?f



 the hair The ?rst gram of animals was treated during
- 3.513-.-. . .. .  a 
(J) mud,? mgnu-phalol  during- 




 
   
 
 
    
           

-. swam?  . -

.   

  ?d?hlompmc?ua acid, ??olWaoh and

  (kl-?60. were Wind repeatedly to rat. for 60 
-    applications in an ?wt to dates-aim  Inimls prior b19911



  
   
 
  


.411 . - 
-.- . . - .- . I - .- A- - finIf-?In Wt: I51 - ?9 mam?an(mum . .I

- ..--. . . .. .n . 
gj?. . 
. .1 - . u-hi?: I
.. I 

. 


   

I the skin  m; (In) mun subjects with apparently norm].

 

II I I fur mind of 27 days.
 gas. mm: m, (m1.11germ?) 413 ?mu; - .

 th?m?ml??md ?hgs-f??o lppliod    a:

w' -:

  . If  --  ml: organ 'mum of h?mn subject- (tm in ash 
   with uppnruntly normal skin, for a period or 59 dun. 2.14.5 




2
1.

  
   
  

 was Ian-tho ml manyInna-.1? \l?TtF*xaaq?hm?" AJrn-v?


.. ?if  ..- . ..-.. -

I r. -- 'vu- "'rr r- Iram'i?l?n "In! two ?5131-15 fwibh?'?aw?m?mu ?Mm99:. T91, "Moo-

 appu?-a?dh" to  Skit; or  lulu rats
7  5% #1 awn.Loh'" 
. 



  

   mm'? 1?  4'
I:  9f  phan- o:  63rd@2108  in wad  a tam. _npplicu?

  



 

urn-Emulation  a! mug21;.

    
   
  
 
  
 
     

 

       

  
  

 

. . I -  II IIit.? ..  - - -Iv?us 
. - . .-,
calca' for a pried

-, -.
?l I .
ma

 

Mn.  .an1 2:75-
. l' 'r?e "z a "5 --  
?alv-IJI 

   

J-qq _i-q-un- --..M -- I .. . -
guy. I 
chug-.- 4'mla rats to: perimh up to 31mm. failed

Ju- 
3pm..?  . 
39-'25F'rf?t-.-I - J. ..

oz? Fibula-51m managing an; lam cr other
mumskint- ll Wamm'pmad, prudde no

31mm or pmmam chm-m 1n the ma. 15153
 .    

- If?, . . .3 - i  
?pm-1' 1" PW 
 _mu a warm ..-- I II . 
tun-MacI.I.6. the application or 57-; column a: ?when 1031;" to human skin  

5%

"pm-10d produced no clinical  pgt?ologiml .-
aw?.  In?euum and mama-Jun gmuem   . 
I1:  lid."- 

It.?  

1- - - .
J?s ?lth. . II,
-

  
     

   
      
 
 
 
 
 
 
  
  
   
 
   
 

    

hiv-  imd tour (14) of  
31th thin  than such as  marina  1011;". 
   ?13mm mditiana mloyad. {In rats, 372



     
  

.  Qaiiomh Human 98-1-60W tam. 31 than. - . . 
mlagtreet, and 1-5 or ?iiz'insa' aka>111 9?er 1  1.
?f ?mt. . - c:
new; mug-muons, 1.abt?.? - . an". Vu?- II
- - "-272 -  L2. (it.

 .-
.. -

1 9? 24115-1? 1953.534 Ml?catim.  

i.

.- 1,39;  tricl?nroinuoh_ 5:33  2;:5335 j. -i

 

,Iiin?plin.  ?atw- ?no clinical or pnunlagi?gal -

11;. . 12,4?.   
.- --

?hl?hw 

th- pix-mums which 1m ?ht m. gunman: 



 .   phi  mulmt  1hHmm? "tau-.9- 
1:25?. 



   
  
 
      
 

if; ja- smut. Bum prunmt qua-um to be mind in non?Jay i-A - [u??ywr?h?a-uq?? 
div-"15r- vim . - .-. . . -.- - . . . KIA. the rat an limx'iato    ?hm.

If}; .v I automates-La per dose applied to at  

mus1'1. 2 1-. . .
23mm not other mm a! entry into the body he considered in

.Iln?gr-vi- afi- uf31h: pummel! at  of       
 -  I- . 

 

 
   

 mm?;     w, 
  I   to this ant-chin which:   
of ma imam m?vn'rrawp'  

- 

$15.33!; I?kin   a?de with
?1m.? 9w   ?cumming . -.-114% *3 - 
m.  ?mitt: MW Manta comm u. and -m -- 
adiabgum-d,- - .
.b-Iu. I.

   

.   .-  

sph? 5- - - nth-J-ln' .. I .- . IIhi.   hump.

Arm-7.11.. on-..-. '1 .-
a
I-

   
 

.uf. - 
. -.. - .qS?n.11., 

-. rbk- 's gm?. 'th  

I mainly may]? upon_ tamyualy dawlpp, arI I111 dash than my__

  

:71 fomm?ar inflammation gin the dog or at;  
I -- I It. would be Vol.1 to consider alnq mains umIin I
in which than is an abunchmo  uhlm    I
 . . .  - - Luv-1.-

such a: sit) in the ?xterm; ?awry-W31" tho am: {not
  .,   7th-  at the ?rich #11011 In why-ad hy am and - 
was). ma mm? or thin Harm'-   3'  
-. .

337m" "W'Wnamwmw; 

 
 
    
  
   


.I.. .. 
I

.15.-- (an.--.. nun 

 l?  I 2. Since that de?es or mkriala were unwind in mam 3mm

   
 
  
  
   
   
 
  
  

1 ?mgbaut egch max-mt, the dam/Hum r.qu 

 saw. in 'm . 
   



. 

 -I-II . -
-


Mac-?.-.. .- --
"3'53" -- $5 -

 Man's    
II in: with Md tau-em11m? in 0.1. 

1'  mm inmi?rleiant th  

  in hm-   mm tic-uagar'irr.--I.. - I. .-.. -. .131  . Inf.-.- --  .  - .. - .I

I. II

?5

  



   
 

   

rzf?i?g??v? .i?e  "av?5: a: 1:ll-l 
-, - .?IuI-lI-tzv-r-u-w-?- I-- r" 

   

.-


1


no  Since in an naryr?Pd 1?
u- - - --

A12.  

 ?n material ta tho errant-ad cram  tbs.

  

in 

mm should be seriously was.de in this summation. In

-.- I- Wm? Maugatmm'mmoum arm?   
. . and manual  .1: 
-I-:Wem-m? 144-4l-?a
r. . d-l-?-.-- ?aa. htasrwg-z?nm?bnm 
I  f?m?-mWh-?I ?M?vf-?iw I sum-n - -- - v-

?inI II: I .. . .-- -..I ..

      

I .. 'Im-gm-uwkrM-?II.. 7 . - -  I II.-.I..Helmmuu- - . ,4 ?ggp?? .MGWI 13.83" Hg?u?dlg?xh1_
..--  
4I.. Iz. ..Ifir
. 
?51-?..515121.15. 

.mail: that cutaneous absorption of smegma under upod- 
 Iknantal noudi?nm my be ?uted. To entrance nah absorption

  

  ?r'?mcuurtg  tag"  
 $31-09  

1'  r' .-
..Lr I 
?Wax-1A1.4. .1
. . 

  

. 


Eryn-t: 13.3.. vi- - -  ;?i?ftti' I  Jrpoufble?thnt?mn? or ill of {the fur {13 W05
'9 =4 

v.
 51- 



  

iris-H- 

4.

w. gas-ii?am?? 5&5;     he}

        

 
          
  
   

333:"? unit awn-pain. We? since the m?ornp?u?ini
 f; mmu?'rmlta mm mm. manna-ha,  thin 
5 Iu Inn. as tho mink- of the ?mum,  I
I   5'1"  3? ?We ?Mam  . -. 

     

  
 
 

   
     

.- . .. 
phuo an.
- -.  '.tut~37 ?Riki-?529v: - z. . 

   bum.  
I twin-I'M Iho' rim-954   

 

  

9.15:1: gunIn?. Id.

. --.  Juana; mime; Mental-.-..--
-. 

 
      
  
    



 

 
 

       



  

 
 

. 

.

. nativity should be carried out. aim;qust in  direc-

    15% m- 

 j; 7  .1193 men arise currently in ma 2.h.5-T donpi
I -    . 

6r 
I- 

of tho menus and rigid Wm mm?Mouton that newsman are still being: and that the patsntinl
- 1P157
- - 
I hard 1: still promt'r?ril-Wmm' 
- 1'43, -- 
I I 
ii-??vwr  M. . .-. 
zit-Ex 1227"Ti-.1- 023d 


.. V.

I 

- .. 
n- 
a 

     ?gurative; ?mforo; that a mutant-1m of .
{All   gamma.? or ?mm-3cm? and a detailed mathrise to the ?mm-n out, 

  
   
  
   
    
        
 

I or abjontifpa o?n

TDDLIQL Moo

Hati?nda q! stuck? must be _davalnped before the

 

-      
?bolppranohad. It?nmu?romomhimduk?lma mime of

  mama .-amm;a as "?sh 3:15am m. I
 a; the suspected mm,'poaa1bla.   requira 

W?mum in 

 _r  hailed mm: mm ?rwtiuna This study
in turns of not last a ?rag-- to five-gear" 

.133h?ws1.1. ~11 . ..-.  . .
a! 351-, p? . - -. - ug?fp_ I$31.4.  ?asmar-w'r_me; tea 3w.  1.1}ij- 133?1'w'?qlmh?? 



    
 


-. 



. - 


my"; -.-..-.- 

. -  
- --. 4-. - -- . .-. .13..-. 1' .- .-
9-: - .I-u 
7. - I . 
 m-md-muudc ?Tenth 5.If43?. I. . is 4- w'or. a $-high combationa and 13an cutaneous arms to a

 I (If  mimln including 505; cm, rats, etc.
    the cutaneous and mums effect: or lam aoimgana

 aha-dual ante-n1: from the $11.-.-J-. - 

   over an extended period by inhalation and a, m1

roo?ng. A variety of minute]. munch should be mod. - it);
 a. the sebaceous-rich skin of tin external  and

I.

. - 

{?rthPr.- . -.  
I - -- ?gamut. magnum in?rm .9: _mm amt-.?4.x Ava-5:54:uyu'au-prwa-h" - taut-W?W' van.?- --?mlu Iv-Mo?

  
  
    
  
 
    
  
    
   
   
   
       
   
     
    
  
 
    
  
  
  
    
     

. 563g?? ?Kg 33.3023 n3 3 5%

was; M00

 

.a

. .I. .I.. . .I .  
Elwinan"? .I I.- II. 

  Farr.      . 
. .

.-
r.l. .. 



  

 . 933?.  gauge. man 83353?. FEE. . on .56.I.Ialt?II .

.  Em. on maggots. 95.48..I.}..Iiullk . 55.5.1 I0. .I@5336363."3an .rc: 3888a.
.. . . {.933an.gguiggji.I-.- I.. ..I 
I.  .I.. a. waili?if: ?(Iii-I. 

.I.Wain: . z.
..I 21   

     
  
   
   
 
  

   
 
  

.. . LiImquA-fE?Jz?u .r
.f ..oumpg? .

Hug

   
  
  
   
      
   

 . ButTlh'r? 1?1 t. u.
. I. unfit ..

Fur. 


    

 

  
 
   
   
 
  

.fni . .. .21.:11til. 
u. A 



. 
I - 

       
     
  

gun-4 



.. rll.u JI

4&2: .5 is. 4% 

 .43rnur
. . 


. .rt: 

 . .
. .. . . 11.. . a .
{litanlhl at.I.. . . .I.IIhunini .I
u- .I . Jill. .II . I?ll . . I I .I.I...I.II1IILII1I  .. . Ivl .I.I..I.. . .I.. I I I I .I.. I. .. . .
I.I.IllIrlI .fl IIll .. It.? I flu.I.. I. I I. .I .  51.. I .IDOW 751763

mutant. to ?ll.

 or [Input-II A

31:15.39: am.
Li; 

aston- of 2,11

 

.

. .2:
If 

mil

IHII II. 
.. .

I .I-
I 

:Innh?k?itamff 

I . 
.I. ..

 

lawman I
Platim_

Sift-J 
Elk": 

l?u'fu Iv

Mac13-Janu-
I 19911
My

 

34:1", 


 
th of Lu.  -
 Applian-
 Mom

.
{my -  
Applied flatl-

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

  

. . . In. .5

1 I11I?ll(.2215 1.1.1.145? (5'ntli. [II-?.IIT .-

.Iull.

 

 

 

     


. . ..

  

 

C. 4.211212%  .. 

 n 1  I. Jillfury-34.324 .l .4118}. . 

. .

J. Armani.)

 

'Jto-nuoa rum
man noIl..i..v \xli!

 

 

 

0? 1.I. .
.J . I

A.- 

 

 

 

 

1.31.? .Jll. .1. II 
LI. 1 Lilli-gr?

I

 

.7111?.

 

 

 

 

 





 

 

15184

 



 

 

x1. I:


..33:1 .. 

til.

 

 

 

 

 

 

 

 

 

 

?5

w.
0
mu?an {lino

 

Luau 
. I 



 

-

 

 

..
. IliumIla-ll
.

(ItIl. l. 

      

     

   

.Ef?tvdihu?il?  
EJ.U!l.rluluall I

Siknfri .. 
.
.

 
 



.. angina ..  a
ht: . vi??vtuif . . 1.1.. I .?Irll (bulgingtaught ..?ibw?dlnd?umhbg
. . i I .. .1: lily-.I

. .- In
grd?itn?tfqh??m?i 
. . . . . .?ulr

inductIll-I}

. 1 .. 
. (ganglalhl?di. . (.1111!
. . . .. pill .. .


- .I rll. I I. all 



 

 

 

 

 

 

 

 

 

 

4 .

J.- 
willGIL 

 

 

. . .
(run?alt. .
  

a
I. I-9.1.11.1.4.1.453:

1

 



 


. .

 

 

 

?4


gunk.lhlilr. u. .0 .

- 11.1Ill. .4 l..lul.?ll.u.l.luv

 

 

 
 

.. .I - ?lly??1.11
I'ill?vun

II:

 

 

 

 . In?.3.3.. . imam?.uni-?n?? 1.77.4.-.I..IVE 

?Pathogenesis of Chemically Induced Acne? 



The objectives of this study are: to compare quantitatively the responses
- 
of human skin with rabbit skin to chemical acneigens, and (2), to study in detail 
the structural and bacteriologic changes which occur during the CD
. :3

evolution of experimentally induced human disease.
a. The subjects will be healthy adult male prisoners -- inmates of the
Philadelphia County Prison at Holmesburg, Philadelphia.
b. acneigens - 2 substances of different potency will be evaluated:
1. ether
2. tetrachlorodibenzol dioxane
c. procedures: - The initial studies will be carried out on the back
applying . 05 ml. of the aneigen in a suitable solvent to a 2? square of skin. The
area will be covered with a square of gauze held in place by strips of adhesive.
The applications will be made once daily. The principle variables will be concen-
tration and total number of applications. The aim here will be to establish thresh-
old concentrations for various numbers of exposure. This will require. of course,
a considerable number?sof range finding experiments.
Both anergens will be applied to the skin of the same subject in the
final tests. Moreover. tests of liver function will be included as follows: SCOT,

BSP. Bilirubin, Alkaline Phosphatase, and Cephalin Flocculation.

16189

d. fathogenesis - The skin will be examined for clinical signs of change.

 

At appropriate times, punch bi0psies'will be removed and stained as follows:
8: E, Orcein-Giemsa. Hale, Mallory and Alkaline Phosphatase.
These procedures will give detailed information of the effect on vasculature, the

of mucopolysaccharides, collagen and elastin, and the general archi-

tecture. Enzyme content will be evaluated by histochemical procedures for
ATP'ase and succinic dehydrogenase. As regards three dimensional views of

the lesions, the epithelium will be separated by sodium bromide and mounted in

tact as a sheet.

e. bacteriology - Cornedones will be removed at various stages and the

 

contents sampled qualitatively and quantitatively for bacterial contents. For
comparison, a similar study will be conducted on comedones derived from patients
with acne vulgaris. Lesions which become inflammed will be sampled for their

bacterial content, either by biopsy or by aspirating i'luid.

f. horny layer - The horny layer from treated sites will be separated as

a sheet and studied structurally, physically and chemically. Values for normal
horny layer have been obtained. We wish to know whether hyperkeratotic samples
represent an increase-"in mitotic activity or whether the epidermis is pathologically
altered at the same time. The rate of desquamation of the horny layer will be
estimated by measuring the time required for the disappearance of a ?uorescent

dye which has been appropriately usedto stain the above horny layer.

-2-

16170-

CLOVER LABORATORI Es. INC.
P. 0. 7686_
PHILADELPHIA I. 

EVtiontcu 6-3459

December 21, 1964

8996151. MW

To: Dr. V. K. Rowe 
Biochemical Research Laboratory 
1701 Building
The Dow Chemical Company
Midland, Michigan

From Albert M. Kligman, M. D.
Clover Laboratories, Inc. 
P. O. Box 7686
Philadelphia. Pa.

For: Study with Acnnigenic substances

 

 

 

Help and Overhead at Prison - 000. 00

Subiects: .
Approx. 100, average cost $15.00

per subject 1, 500. 00

Bacteriology l, 000. 00

Histology and Histochemistry 000. 00
Miscellaneous: I

Materials, Photos. Hormones, etc. 500.00

Total 000. 00

University Overhead 20% 200. 00

 

TOTAL $7.200. 00

 

 

Grant to be made out to Trustees of The University of 

15171

g. Prevention and Treatment 5 It is known that x-rays. estrogens, and

keratolytic agents are helpful in acne vulgaris. The beneficial effects of these

modalities will be evaluated.

h. miscellaneous studies Limited attempts will be made to evaluate the

effect of age, sex, and body area on the develoPment of experimental acne.

Re spe ctfull ubmitted.

Albert M. Kligman. M.D.



I 6996M MOO

. - . -
Jab 

18172

 4? . 

that have had implants.
Department of Agriculture told Environment
that residues of DES are not ordinarily found
in animals with implants because the levels of
DES are below the sensitivity of the analytical
techniques used, but that there were

A spokesman for the

undoubtedly some residues. The Spokesman
also explained that the small farmer will be
hurt by a partial ban on DES because whereas
large farm operations can simply switch to
implants, the small farmer does not have the
equipment needed for implantation.

A SHARP INCREASE in the incidence of a
particular type of liver cancer in North
Vietnam may be related to herbicide spraying
programs, according to Dr. Ton Th'at Tung of
the University of Hanoi. Dr. George Perera
who visited Dr. Tung on behalf of the
American Friends Service Committee told
Environment that Dr. Tung had seen a greater
than five-fold increase in liver hepatoma
cases between? the periods 1955?61 and
1962-66. Perera said that Dr. Tung suspects
that dioxin, a contaminant of the herbicide
2, which causes birth defects, can
cause liver cancer. Human liver samples
suspected of containing small amounts of
dioxin have been sent to the U.S. for
analysis, but techniques to test for low
quantities of the chemical are still in the
developmental stage. Although no herbicide
spraying was conducted over North Vietnam,
Dr. Tung claims that there is a great deal of
movement between North and South, so that
his patients in Hanoi may have been affected
by in the South.

is again in the courts. A federal
district court Judge in Arkansas issued a
preliminary injunction June 23 against the.
Environmental Protection Agency (EPA) plan to
hold' public hearings concerning the fate of the
registration of the herbicide for use on rice
crops. In a case brought by Dow Chemical
Company, a manufacturer of the herbicitE.-, the
Judge ruled that Willi-mi Ruckeishaus,
administrator of EPA, must make a clearrul
decision on whether or not to ban 2,4, for
the spec1fied use beinre hearings are hi-l'i.
Last summer Ruckelshaus fnnrul that 

  
 
 
  



he had insufficient evidence to make a final 
decision and ., -'dered a hearing. A spokesman.
for EPA explained to Environment that the
administrator's order was sufficient, and said
that a notice of appeal had been filed. When
asked if Dow was using as a test
case, a spokesman for the company told
Environment that "We've got to stop this
ridiculous mess somewhere; who else is going
to stand up in a case like this?"

A CORRELATION may exist between air
pollution and what is knoWn as the sudden
infant death in which an
apparently healthy infant is found dead in

its crib. Preliminary analyses made by Dr.
Bertram Carnow of the University of Illinois
Medical School showed that when sulfur
dioxide and particulate air levels were high,
there was an associated rise in the incidence
of sudden infant deaths. However, Dr.
Carnow told Environment that further analyses
have yet to back up the preliminary work.

Dr. Carnow theorizes that the sudden deaths
are actually due to a constriction of the throat
opening caused by acute tracheobronchitis,
which is aggravated by high air pollution
levels.

A NEW PRICING regulation by the Federal
Power Commission (FPO) may spur exploration
and development of new natural gas resources
in the U.S. The FPC, which now regulates
gas prices, will allow gas producers to sell
in the interstate market without being 7
restricted to current area price ceilings at the
wellhead, reports Coal News, August 4.-
However, the new ruling applies only to gas 
from reserves discovered after April 6 or gas
diverted from intrastate markets. The
Commission retained the right to ensure that
gas prices will remain "in the public interest."
Natural gas, one of the cleanest fuels, has
been in short supply, a_fact which Coal News
says is attributable to "restrictive FPC price
regulation of gas at the wellhead. 

A NEW WAY of controlling mosquitoes is being
studied by Clyde Umphlett of Clemson
University. A report in BioScience, luly,
states that Umphlett is working with a
microscopic fungus that attacks-and kills
mosquito larvae. If the fungus is found to be
without unacceptable side effects, it could
eliminate the need for spraying pesticides or
oil on mosquito breeding grounds. The fungus
is extremely effective on several Species of

 

Awe. 

-

m,

24

the report of an advisory committee 

H-



 

Bantam/toe 

S99IOONH



Vol H, No 7

3

?~1pr-

MIDLAND
June 21, 1968

H. Ferguson

M. Wiltse
J. R. Ridner
C. A. Bryant
0. H. Hammer
. . . a son
W. J. McCoy

CC:

'Your letter to Andy Watson has been passed to me for ans?

wering.

Enclosed are several pieces of literature_dealing with the

1930

TFEIEE 



JUN 2 1 1953

.BIOPRODUCTS DEPT.





ZIQIBEMOG

?toxicological aspects of silvex.

A good summary of the effects
on fish has been assembled by

The summary by Mullison lists

could supply if you feel this

of silvex ester and salts
Hammer.

several references that we
is necessary. -

'The size of the market you mention in your letter was
a bit surprising but perhaps this includes some other 

areas than New Zealand.

I have sent a copy of this letter to W. J. McCoy.

He will

advise you on his thoughts regarding the world market for 

silvex.

Let us know if we can be of further help.

H. Davidson

Plant Science Research and Development

Agricultural Products Center

bk

encl

. . 18174

 I ad 

arranged for a Joint conference between 

57 the USDA
es of
-5 Biological and Physical Sciences, FDA. Attending th n2 wereg;

2008

NATIONAL AGRICULTURAL CHEMICALS ASSOCIATION



ngice Correspondence

 

. -. I If}  n. I
Mr. Hitchner   
JV 27 ..
FROM: Jack Dreessen 

 

July 28 Conference with 

SUBJECT: 
- 2,h-D and 2,h,5-T on grazing July 28 in the office of Mr. R. S. Roe, Direct

moo Haller_- Agricultural.Research Service,
Ennis,-Jr. - Weed Sections, USDA

Shaw - weed Sections, USDA 
Klingman, Weed Sections USDA .e . i - ..
Coyne - Pesticide Regulation Section, USDA 
. Walker - Pesticide Regulation Section, USDA
Ligon - Pesticide_Regulation Section, USDA
Cummings - Pesticide Regulation Section, USDA

S. Roe - .

J. Lehman - FDA

Vbrhes - FDA 

Jo   

A. Noons - NAG

Dreessen - NAC

I



ici .



901990?

a-

The writer had met with?Dra w. B. Ennis, Jr., in.advance of the
meeting and together we worked out a suggested agenda which in turn

5 was forwarded to all participants by Dr. Heller who presided at the

.31-conference.
Ennis and myself.

1.

The following is the suggested agenda as developed by

Purpose of Meeting -_$g_detergine the information needed to con-
tinue recommendations and 

pasturee;and rangelands,

?ume 

 

H. L. Heller (5 min.)
Status of registration of 2,h-D and 2,h,5-T on grazing lands.
J. T. Coyne, PPCD, ARSG min.)

What industry is doing to comply with Miller Amendment with
reference to use of 2,h7D and on pastures and rangelands.

Jack Dreessen, NACA (5 min.)
Necessity for continuing uses of 2,h?D.
- - D. L. Klingman,ARS, (5 min.)

Analysis of existing data on residues of 2,h-D, and a,
MCPA when used on grazing lands. i?gi?

J. T. Coyne, PPCD, ABS (5 min.)

u.

1


'I-l


 a? 

4r?)

7.

- 2 

6. An?lgsis of existing data on the pharmacology of and
2, - .

J. Lehman, FDA (5 min 

Discussion of questions raised concerning registration.

On June__v9,r_,l_955, Mr. E._I_oynn_wrote to Mr. w. B. Rankin



I 'that"the bulk of the 

?year is made.

following"3xcerpts are taken from that letter:

am writing to be sure that Mr. Sunderland and I have
a clear understanding of the conclusions reached with
respect to 2,h-D and the related compounds in_our meet3.7.9? mm 3      .24"191.9 .af 1 ensth the #0513351? 
2,u-D and opinion

til-rears: . cam-.1 as._.out__1.n 
Stock gratinggonwsuchmlandsware?not slaughtered?gor
months later. A high percentagenof?this stock
after they are removed from the range and
before they are_slaughtered. Thus it did not seem
likely to us that early transient exposure to these
chemicals would constitute a residue problem in stock 
slaughtered several months later. You took this 7 03
matter up with your colleagues and gave us the opinion

that it appeared there_was_no possibility of auresidue
problem undermthese circumstances and that you would-

not askfus to_obtain data on this particular problem.?

Mr. Rankin replied to Mr. Lynn's letter on June 15 as
follows:

"This acknowledges receipt of your letter of June 9, 1955
which reflects accurately our understanding of the -
questions discussed at our conference on June 3.

"In reading the letter I realize that there is one
additional comment which needs to be made about the
problem of?residues of herbicides on forage. _??hherbi:
cide_residues_are present on forage, other than_alfalfa,
which moves 1n_interstate commerce,mpn baled hay for"
exampletholerances or exemptions would_need to be?
'established for them.f - - 

sunny .1
I .muw?a?u h. -

 

?he situation_ooncerning 
may be grazing have'not_changed,
since_l955, the normally
applied early in the grazing season, application rates
one'applicition_per
If the same conclusions he?drawnmtoday as
in'1955 then there is no possibility of a residue problem
under these circumstances and recommendations can be con-
1nqu 0 i

How much and what kind of data are needed to determine
whether there are residues in milk? 

75'

h.



 



-

- 3 -

How sensitive a method is required in running residue


'i studies on milk? 'Will data obtained by chemical, bio- 

logical and radiological methods be evaluated on an equal'f 4 ml?

basis? . 

In a letter dated April 12, 1955, from w. s. Rankin to
Mr. L. S. Hitchner the following statements were made:

T"The toxicity data on 2,h-D would not in our opinion 

Justify the establishment of a tolerance higher than . ru?s

parts per million for this herbicideaa?rti?isr?i??ir   .. I.
"The method used to determine residues of 2,h-D should -
detect residues of 0.5 and at a-level of should
be accurate to plus or_minus_0.5 1 

Can data accumulated on residues from~ one ester be applied()
-to other esters?_ one salt to another? . i

..L



If residues are found, are there adequate pharmacological 'gg
data to establish a tolerance for milk? I 
If residues are found to occur in milk from animals grazing 00

treated forage and it is found that these residues are no 
longer prevalent within number of days after treatment ?a
will registration be continued by including on the label

a statement to the effect "Do not allow milk animals_to

graze on pastures treated with 2,h-D within
-days after treatment." .

 

8.  Sumarization

'The various items on_the agenda were discussed and the following are

my impressions as to statements made by various persons attending

the conference. -

1. Dr. Heller merely expressed a desire to try to develOp what is
required for the Department of Agriculture to continue recommending
the use of 2,h-D and 2,h,5-T on pastures and range lands. 

2. Hr. Coyne stated that the status of registration of 2,h-D and
on grazing lands has not changed as far as his section was
concerned. He indicated that letters which have been sent to
registrants in reference to clearing the use of 2,h-D on range and
pasture lands had been mailed to put the Industry on notice that
such uses had not been cleared in the Section's opinion but that
registrations were not being cancelled. It was his Opinion that the
basic producers' action in removing the use of 2,h-D on range and
pasture lands had been done at the Industry's own volition. Es went
on to use of 
continue is 
indicated by administrative the
ing these uses ,w?i-twwould . .be
aocop?ed. Attached as exhibit 1 is a copy of a letter to one of our
men or ?ompanies similar to those that have-been sent to other
registrants of 2,h-D in reference to this problem.

181?7_'

 I
-1..-

3. I commented that the Industry removed the use of 2,h-D on 

. pasture and range lands from their labels based on letters the reg-  I
-ou . istrants had received from USDA and which were commented oh by 
Hr. Coyne. I stated that it was the registrant's opinion that the
Ln?s. cost of collecting data to clear the suggested uses under the Miller
hater 'Amendment were not Justified on the part of Industry based on the
54?; expected cost of collecting such data versus the expected profit

jgw?f- from the continued sale of 2,h-D for such uses. These statements
,Tgf'3_ were made based on discussions at a meeting of the Herbicide Tech-
nical Subcommittee last fall and telephone conversations with

in?g? -members of the Subcommittee prior to the conference?1.13:3; . .

necessitlmforuthe continued use ofmzih,D and grazing
fiW?ai lands. A Exhibit?g;
g?au?g Dr. Klingman's statement brought to the attention?of EDA the neces-
city for continuing the use of 2,h-D on pasture lands._

staff who reported'that the had insufficient data to conclusively
determine whether 2,h-D, 2, and MCPA leave residues when used co
on grazing lands. They also stated that they did not have conclu- it
sive data as to whether any of these products were transmitted to 
meat and/or milk.-

~9r37; 5. ?Hr. Coyne tororted this item to Dr. Walker and others of his? 5%
ex.


. 6. Dr. Lehman stated that if residues of 2,h?D and/or 2,h,5-T occur
. in meat or milk that further pharmacological studies would have to
be conducted with these herbicides. It was pointed out by repre-
sentatives of FDA that if residues of any chemical or its metabolites
are present, that pharmacology of these and products would have to
be determined before FDA could pass on their safety. 

7.. The various topics listed under item 7 were covered during the
course of the conference and definite answers were not obtained in
all cases. 

Ya) It was generally agreed that the situation concerning 2,h-D,
and MCPA applications where beef cattle may be grazing have
not changed since 1955 (refer to this item in the above agenda).
However, based on previous statements, the Pesticide Regulation
Section had continuously accepted these chemicals for this use and
could see no reason why such registration should not be continued.

This question was not adequately answered.

No specific answer was given to the question raised in this
item. FDA indicated that the nature of residues, pharmacology or
residual material and many other factors would have to be considered
before the sensitivity of a method could be passed upon.

FDA indicated that they were of the opinion that the various
esters and amine salts should be considered as different chemicals
?~and that if data or pharmacology on residues is accumulated for one
or more esters enough work should be done to indicate that this same
data could be translated to other formulations. Indications are
that FDA and the Pesticide Regulation Section would accept a limited 

 ambunt of data on some esters and amine salts if it can be shown 
"that their properties are similar to other formulations on 
more complete data has been accumulated. I -

.  

which

The answer to this question was No.

It seemed to be the opinion of the group that label state-

I'li.ments to the effect "Do not allow milk animals to graze on pastures

treated with 2,h?D within days after treatment" would be

?1.acceptable provided sufficient data had been accumulated to con-

which would result in no residues occuring in milk. 1

:ZThe above represents some of the answers to specific questions

'5 {raised in the suggested agenda.
2 quite positive in stating that they did not have enough pharma-
a -Ww.cological data concerning 2,h-D to establish tolerances should a
-residues be found in meat or milk. - '3 

clusively indicate the interval between application and grazing
   -

. 

I would like to emphasize that FDA

9&8948 M00

mm

Jack Dreessen

-.  




Us

#43153.
..



i'f?iv

2037

KURON

TOXICOLOGICAL INFORMATION AND SUGGESTIONS REGARDING U.
TREATMENT FOR PERSONS SWALLOWING KURON (3
a: 

FIRST AID to -
c: to
Call a physician immediately. Keep the person warm. :3 9?
l-?-aa
Do not attempt to induce vomiting. I: a;

\l

To THE PHYSICIAN

 

Kurod?)contains 65.3% of 

propionic acid, propylene glycol (CBHGO to 0 H1803) butyl ether

9
esters and 35.5% of inert petroleum oils and emulsifying agents.

Kuron has a low acute oral toxicity. The LD values

50
for various animal species are: rat, 1070 mg/kg; guinea pig,
850 mg/kg; rabbit, 850 mg/kg; mouse, Elho mg/kg; and chicks,
2000 mg/kg.

The advisability of attempting to empty the stomach by
emesis or gastric lavage is controversial since aspiration of a
petroleum solvent is to be carefully avoided. Respiratory embar?
rassment caused by pulmonary edema and pneumonia is the primary
danger from swallowing the petroleum solvent. In such a case, the
administration of oxygen probably will be beneficial., Chemotherapy
would have no effect on the so-called "chemical pneumonia" but it I
may have some beneficial effect in preventing or controlling
possible subsequent bacterial invasion.

Depression of the central nervous system may also be a

problem. 011 laxatives other than mineral 011 should be avoided.

16180



- 2 

Liver damage may be minimized by placing the patient on
a diet low in fat and high in protein. The usefulness of lipo-
tropic drugs has not been investigated. Kidney involvement is
rarely sufficient to merit special treatment and for those rare
cases where the kidneys are seriously involved, the treatment

should be the same as for toxic nephritis_or other etiology.

NOTICE

nweaemoo

This information is given in good faith, but while it is

believed to be correct, no warranty is made.
NOTE: In case~of any questions, call The Dow Chemical Company at
Midland, Michigan, on phone
I MElrose 6-2376 (Toxicology Laboratory)
MElrose (Medical Department)

Biochemical Research Laboratory
The Dow Chemical Company
May 28. 1959

2038

2.30a237ul

Biochemical Research Laboratory

  
 

The Dow Chemical Company E3
RESULTS OF RANGE FINDING TOXICOLOGICAL File 
TESTS ON AGRICULTURAL CHEMICAL FORMU- No.. Sample 1 3: Q3
LATION M-2121 CONTAINING 50% SILVEX Chg. 12714-26 'z 
EQUIVALENT . Ref. GH3-677-137 to
Sub.by G. Scoles ?'95
Signed 7 Cf NAZI, Rept.by K. J. Olson 
.
Checked a?
DISTRIBUTION
The Dow Chemical Col, Midland Midland Division Texas Division
R. Boundy Tisdale D. J. Kilian
H. H. McIntyre E. N. Luce R. w. Colby
R. P. Perkins L. M. Greene
A. Hart 3. A. Shrader Western Division
N. Wright w. J. Falkenstein CRI
G. E. (2) - D. Elshere
G. D. Jones Louisiana Division J. F. Kagy
K. Barrons (2) CRI
G. Scoles James River Division

CRI

18182

THIS REPORT IS THE PROPERTY OF THE DOW CHEMICAL COMPANY

Biochemical Research Laboratory

The Dow Chemical Company

RESULTS OF RANGE FINDING TOXICOLOGICAL TESTS 0N
AGRICULTURAL CHEMICAL FORMULATION M-2121
CONTAINING 50% SILVEX EQUIVALENT

PROBLEM

LON-IZEMOO

A sample of M-2121 was submitted to the Biochemical Research Laboratory
for toxicological testing and definition of industrial handling hazards.
The material is a pelletized formulation containing Silvex Acid and
Silvex Potassium Salt on Attaclay. The formulation is designed for use
as a brush?killer in the undiluted dry granular form.

CONCLUSIONS

M-2121 possesses a low acute oral toxicity and is only irri?
tating to the as well as to intact and abraded skin. Excessive
skin exposure to the moist material, particularly if confined on the
surface, might result in some skin redness. Such exposure should be
avoided. It is anticipated that no problems should arise from handling
the subject material under conditions where reasonable care and clean?
liness are practiced.

SUMMARY OF RANGE FINDING TOXICOLOGICAL DATA
Acute Oral Toxicity

 

 

. Dose No.Died
Animal Preparation Fed (g/kg) No. Fed Response-Remarks

Rat 5% in corn 011 0.50 Animal died four days
after feeding, pathology
ess. negative.

 

 

 

 

 

Rat 5% in corn 011 1.0 0/2 Pathology ess. negative.

Contact Rabbit -

Material Treatment Response-Remarks

Undiluted Washed and 81. conjunctivitis subsided in one week.
unwashed 

 

Skin Contact - Rabbit
Condition No.0f

 

 

 

Material of Skin Appl. Site Response-Remarks

Undiluted Intact 10 Belly No irritation observed.
(Dry)

Undiluted Abraded 3 Belly No irritation observed. 31.
(Dry) scar in 21 days.



 

 

 

normal in 21 days.

Undiluted Abraded 3 Belly Ess. the same as above with s1.
(Wet) burn following 3rd appl. Skin
healed with s1. soar in 21 days.

C3

Skin Contact - Rabbit (Cont'd.) CD
Condition No.0f :5

Material of Skin Appl. Site Beeponse-Remarks Cu
Undiluted Intact 10 Belly 31. to mod. hyperemia with 31. 
(Wet) swelling and exfol. Skin Cc
CD

03

NOTE: When applied wet to the rabbit skin under a bandage, the material
adhered to the skin which probably resulted in the irritation.

Skin Absorption

There is no indication, from the skin irritation tests conducted, that
this material is absorbed through the skin in toxic amounts.

l?l84

FIRST AID MEASURES

 

 

 

 

 

 

 

EYE CONTACT if the eyes are contaminated, they should be flushed immediately Iith copious amounts of flowing for sling;
. 1) minutes.
Medical sttention should he ohtsined if irritation persists or develops after of the eyes. 8
[3 Medical attention should he obtained. 
l:l HEDICAL ATTENTION SHOULD THEN BE OBTAINED DELAY. OJ

SKIN CONTACT Any injuries or irritations which any develop should receive medical attention. 0?3
03
Contaminated clothing and shoes should he removed not tensed until thoroughly clesned. CD
3
Issh contaminsted skin with sosp and plenty of water. (L
clothing. including shoes, should he removed and the effected shin area should be washed thoroughly Iith
soup and plenty of Int!!-
I: Medical attention should then be obtained.
Contaminsted clothing and shoes should not he re-uaed until thoroughly clesned.
All contaminsted clothing, including shoe-?mug be removed immedistely and the affected akin area flushed thoroughly
sith ester from a safety or other suitshle device and clesnsed I'lth soap sod plenty of Inter.
MEDICAL ATTENTION MUST THEN BE OBTAINED AS RAPIDLY AS POSSIBLE.
E3 Contaminsted clothing including shoes. must not he te-used until thorougth cleaned or must he discarded.
lat-[ALAan Cl If person should experience any notice-hie ill effects from breathing the "pot or tunes of this msterisl. medical
attention should'he ohtsined ptomptiy.
If person should be overcome from htesthing this msterisl, he should he removed to fresh sit st.once, he tend: to rest.
hept Inns, and MEDICAL ATTENTION SHOULD BE OBTAINED IMMEDIATELY. If breathing stops. nttificial
respiration should he sdminiatered.
If appreciable amounts of material are s-slloIed. vomiting should be induced by tickling the of the tongue with
the finger or by giving sn emetic such ss 2 tnhlespoonfuls of table salt in 5 glass of warm water. Hedicsl attention
should then he ohtsined.
Cl If the msterial is swallowed. vomiting must be induced by tickling the of the tongue with the finger or by giving
an emetic such as 2 tahlespoonfuls of table salt in glass of warm Inter. MEDICAL ATTENTION SHOULD THEN
BE OBTAINED WITHOUT DELAY
CDMEHTSI
na 



20



A?asiuas: 7;;?37

I??f7' IV. RESPONSE OF HUMANS TO KURON AND OTHER FORMULATIONS
OF SILVEX.
1. Wolf, M. A. 1959, Results of Human Skin Irritation

and Human Skin Sensitization Studies on Kuron.
Dow Internal Report No. BC T2.30-40-3.

Wolf, M. A. 1961, Results of Human Skin Irritation
and Sensitization Tests on Kurosal SL 
Dow Internal Report No. 

c: 

91602VNN

GAELLZ M00 0

18186

 .


..

 

airiz?fil.

?If -

I

5 2717/,  i  


try m'tJt 
('lern 3: lull-T
- -   i'a-aith-MS

lie-n: plan in am: ,tjuy-
ermrn::.t for sen-ml million dul-

lat':; {or in ?ardv-ns,
Ilt?tmitrti; and property they
claim In-
tlim-I?lmlnale 11.4: n! hi-i hh-ldrn.

"In art-lithai \zc (:lxln't'l tn 
Cl.th (iut' ?ficti-
the us. Ser-
vice for damage to the en-
says the 
of a group of Fall resident: claim-
ing damage.

\tln.? li'.
enlists, cu:
national ia-aith to
i started in




'lhe sprays Evil), 2.
and linit .., also (.1133! Sitter-1 

par: of a: mam-3e-

Jnent program to out native .

growth in the l?inzl ltodntain
foothills, says Robert Courtney
of l'lmeaix, 'J?Llito NLHOJLEI 
(Lil supervises.

'J?he Salt irrigation pitiful 
coti:e 
l-rnsh Cu't'il' mere-252:. \rater
Ihrt'.? into reservoirs.

"The spray is definitt-ly de-
signed to lzil! sin til-3 and brush,"
Ct-tii'ilil'jf b'iili, ?51nd 
tie-all must bread-leafed
Illattii?."

ltonieer. nz-rs in nearby lies-mil
Gulch Ktlitl?'i? Canyon rgree
that tin.- spray.tlnes, indeed, 
feet broad-leafed pleats -- not to
mention fruit and shade ire-:3,
birds, animals, people and the
rater supply."

call it ('l'cnto lx'atioz'zal lv?cr-
est) the tort-st,? say: .?Frs.

Leta :limniate the grunxlh 
the and hill the living."
(All are claimiaatid hjtit'uf?r-
bear, and do regulate growth.)

 

She is lira. Sitcom-alt, I
enlisted the help of si-i- 
and a 

if.? 191.: Service 
in with the Salt lli373"" 
1, tilf/Vr/lx?j"  deli/ill

- ill-ml liinnl-r 
 It! t?nillrm,
Minuet-rail L'ntitrmh UH:

chin; thing about 
Ventt?ilius rim-.t-r out ol' 
fut' land
plants hn-aplimhly are hath-as
'lln: ritm?tn oi Lita-t. .?ihun'ra
ham invariant-Ii 

the problem with several 11:0

pie." 
Citizen sentiment 

being compiled by the Go'.

Corannssioa on Arizona tic-an!-
The commission also has ce:

suited sereral L'aiscuity of Art-

zoaa scientists ahoat the
"Nuletiy t-ut CUT. :1 
tize ilL'JIilflL'll'. r' 
cording to Dr. Paul 5.
?The -

PC. i  

JiaHin, 

too i 1211-: 

The same herbicides 
lit-3n used in 
Lara been nae-ti eiL-r

enemy areas. Martin, 
only says:

?At the rate they're .:

spraying, the For-2:21 Sen-:c

I must have tonazl e. ?.'iet Cong in-

i Daring the past several
he has been (olleciing in
the Finals abnormal plants
apparently by herhi<
cities. 'tne scientist also has tat-:-
ea Uit students on field trips to
the area.

Martin feels that the Forest

,Serrice has used a treatment

whose total era-ca; on the earl-
ronmeat are rial-mom. "it may

take us- to find out," he

criticizes. .
Pres-S trill comneat
nume- flu-.1 (ring

a





a matter of too nan-h (lulu; 


a innit) tn 

I, i'JiiEI' "nun 

h-mn Martial

H'llt?t?fi litu 


riagy;w.rth nut




it

,Snming that
Sl
cause
to 
Glut-3 creltliolegist 

Lama 
ill: 0.21' ju'i'l?iz' 
i i' 
-. . . umlile

C-ri'ld' and 

fit: in t.

 

a Lind
3 has quit:- noticeable
"153' 

l. .rc inert- 


may 
icI-L's 
:il l: (1.:
tits-s 223.32?. inn!

an! in 

L'l'll.

lgees-:- an-i 'j-If 


die-:1 daring .li-rze 
many .
0111' HT.

is 

.v 1
ll'i

.lniy, 

 

7? 1r.-
A  A  fl"

refit-ct (ill-
an it 

Another cmit'i't'nul is
?up INA Ut'.
annual the ain'aying. 'l'Lui
committee has KilliluitiL'ti tu lisp. 
Sam Sleiger (it, Ariz.) trim t'i.s.l- -
ed the area last ?to dluCtl'



M.

Lat Service has been lax in as-
iicibicidcs can't
undue tlzimage simply he~
cause they're not suppose-1i to.?
3 lie referred

report by
chemiczd spraying currently is

.515. Robert
cum-'5] on oc-
(unmet-I; the tour
I): at lit-liLL-r Canyua." 'tize
i are ercerllt-s 
23,

l't:

r.c?.e tux-JG;- tu iiy, had
ut' senile-n shut
lo cal, (frinl: or stand
3' L-rungid ia
v.3, lilli.
days

. 
h. In.)

nan-:3 and no lletr


simuz'n top
all their tend-er

leaves in on: dzzy and began to

Cisinlegra'i with hat}: and'
Maniacs fa?iih'J nil. Chit-Lin,

i--ir ll grai:

stand mil up all the
leave-3. 't?iie teal became sic}:



.

I 

 
 
   
    
 
  
   
  
 
  

a fade,

A ilillt: il'ct',

side. On
Lincl:
[fretit'ittal

fruit-3d iuii)?.

think:

.- 511."; 



 

they point to butlluiittng I
pretty 

lh-rayml, Iii-{united ill-ad
a-t-n during: a in
Him) I'tl art-a:
ninl nur-
mal on one side,
no needles on the
Globe garden, Several reins of
le-grcen on one
the other, they were
and decomposed. And

rotted before

pintll'.? ('nlthi Im-
c'rnt vintt in tin-

v. ere chi

(Tenltl natural diseases have
cam-ed all this? 
.1 plant 
nut. Assays have in.
[be ltlEni-J In.? blue, in:

'20-'74





c-unnia'nlmt "Jul.
all MthI luatnm n, hunt: 
tin-an \t'uulti prove
damage.? lat,

hut itil'iJL?
as a:

  
    
   

had cones but

other. In a

?th

Stanley 

5.1) .4, but

he believes it to 1.: unlikely that

any on: virus would
many different plants in the

some ray.

?t?o their mood, the
Forest Sez'rice is so-nen-hat ia-
iiameci by ?hat on: 5
bar termed "not too much rah-l-
pat't of same people

ttitle 0.1 tip:

Gleln.?

Frank!) 
diu-isic-a ran
at 

 

 



?Ellayin folks are a
little scared,? say. .
the Service's
 management

:tiieet so

aft" ment-

 

 

 

A. 

 

He pointed out that ?there?s a
to throw a reg}: aroantl

People

make the

same nt 
tit-ides, herbicides and 
'tize federal gurei?in?nan nonc-

Ilielt-ss, last 
the lii: 0i 


after learning,

lint i: may be dangerous to 

mat tint-53 the Feed
1: or a, (in; lmq: Administratimi esialdciu .x



a "sale legal tolerance in and on
foods,? the US. of

Agriculture \iill the the in!





161$?

 

 

riur 2-. Sonata int-cotigattion,? 
Vice Prcsitlent ll C. LongI..- in?. ?ha-u--w-w rt . . 
2, 4, 531' on food craps cricctit'e
'hume 
In a statementtoo it. Dullaulge,? prastmeittlult Ix]
?dl'lSJl', pomtcd out that. i .3 3 3.3! wt: - i .- '1 
l?tl.c relationship of the cliocls' 3 qr ?Era? :31? - ?1 
Inliorzllory animals I g, ?t .  
i'to effects in man are not culit'c-, .31? - yrs; )5 ?Iiy?xtf?ilE-kq :57 r. 5.: 
fly clear atthis limo",  i. Ally}. ff} I . ..
Not so, says the National 5 I 54.? if?) -
Health i-?oJc-rntion, a. nonprofit nip.? I
foundation with headquarters in?.    ?Inn-fur: . . .- I
l?Jonrovia,CaIit?. Since the {C(lCl-l ?1&3  I I  - it?? 2'53; . r. I
[alien began an investigation last   gran" 
'July of the Globe sprayittg,?itl    51% are: I . -
has ?gathered enough material :112

pt'ojoct in the;
?nals was t-ppi?ovctl by ill-3'
Committee on Pesticirie.
Control, the For?st h33_
"doingtd future herbicide trout-
maul," admitting there has.
"some drift damage.?

do feel that we've- t'clty
tltm'ughly all the.

 

coniplointsf?
gilt'nntm 5::i'dlas 

f23?323?33 vn 21;; mos Fumoxq 

can: 20:: pgnoqs 53.1132; 31:: ?nn; 3:
?2ch ?uid 3131.?! 359-.

quI I
?It. .1 1? r'Ifl.


u-u

E14
(.31:33:) :1


.

.II


ESEQ

ll
0-

At 'swt

ii1:30 Acolouomnag vn ;a Manna: aloud -

to

- 

L-J 

I


no. 5-5?
L.

- wt;
U.

?vwfa.--I..-
'i/Ij AGNEW DOW
MOYER DOW

I 
I AGNEW Dow
MOYER Dow

 

 

 

. - Ur2723

FESTIMONY OF A. T. TALCOTT LifiJam-1Talcott, administrator of Regulations and Labeling in the
Quality Assurance Department of The Dow Chemical Company. I have a Bachelor
of Science degree in chemistry from Alma College, Alma, Michigan, and have
done graduate work at Michigan State University but have no advanced degree.

I joined The Dow Chemical Company, Midland, Michigan, in 1957 as a chemist

09969le M00. 

in their analytical laboratories and moved through a series of technical
promotions in the analytical chemical field. In 1965 I moved to the Labeling/
Product Registration section of the Legal Department, becoming supervisor

of that section in 1975. In 1976 I became manager of Product Safety Compliance
in Dow's Quality Assurance Department, which included the functions of
labeling, material safety data sheets, and product registration. In 1979

I assumed my present position where I am re3ponsible for labeling policies on 
Dow products.

I am a member of the American Conference on Chemical Labeling, the Labeling
Standard Revision Committee of the Chemical Manufacturers Association, a member
or chairman of several committees dealing with regulations and labeling within
the Chemical Specialties Manufacturers Association, a former member of the
Labeling and Precautionary Information Committee of the Chemical Manufacturers
Association, and the Labeling Committee of the National Paint and Coatings
Association. I was also one of two industry representatives participating with
EPA to put together their National Symposium on Pesticide Labeling in 1974.

I will testify on the history and procedures used in the development of
labels by The Dow Chemical Company and to the registration requirements for

labeling used by the Pesticide Registration Division of EPA which was part

of the USDA in 1968 prior to the formation of EPA. I will also testify  -. u-



the relationship of the labeling policies used by Dow and independent standards as

well as how the regulations promulgated by the Pesticide Registration Division
M.

which-apply

n.1r/ i .(p'pu -.:

T3
?in

to the product weed and brush killer.
The procedures used at Dow allow us to get the input from our experts such
as health professionals, technical specialists, and legal and regulatory counsel

in areas.pertinent to such a product.

The labeling used on KURON weed and brusEXRiller in 
mL

?through 1969 was in?part specified in regulations for pesticides which were called.

pea
economic poisons under the statute in effect.at?thatntimei These} equired the

product to bear a label containing the name of the product, the name and address of
the manufacturer or registrant, the net contents, an ingredient statement which
was prescribed in a special manner, a warning or caution statement as appropriate
to regulations prescribing same, and directions for use?-which if complied with,
would be adequate for the protection of the public. These elements of a label were
further prescribed in a certain manner by the regulations and interpretations
promulgated by the Pesticide Registration Division and required a preclearance or
acceptance by that Division before they could be used on the package of the
pesticide. This labeling was also submitted to the state of Arizona and other
states for review as part of the state registration of the product required prior
to sale. 

The product name KURON, the Company name, and certain other elements such
as the registration number required to appear on the label are for the most
part self-explanatory.

For precautionary labeling purposes,pesticides were placed in one of four
different categories based on certain toxicity data that would be in ani al tests

 at:

provided to the Pesticide Registration Divisions These were caliedfcategories

one, two, three, and four; gidh category one being materials that were highly

toxic and requiring the word "poison" and the skull and crossbones to be used on
*Trademark of The Dow Chemical Company

16191?

.- 

an

-3-


the label. The other categories moved in decreasing toxicity to'category four

I a I a
which was for products determined not likely to cause injury under any reasonably
foreseeable conditions of use. Each of the second and third categories were ten?

fold decreases in toxicity from the lower-numbered category.

During this time precautionary statements used on pesticides were set

29969? mm

forth in a document called "Interpretation 18, Revision 2" published in the

Federal Register of March 9, 1962 as an interpretation of 7 CFR, Part 362;

 

There were two ways in which this interpretation was used._ One related to
signal words and statements of hazard that were used on pesticides/Eontaining
specifically?listed active ingredients and the otheraparn set forth general
procedures based on the toxicity categories previously mentioned which was
applicable to KURON weed and brush killer.
. . 

Applying the principles stated in Interpretation 18, one comes up with the

precautionary labeling which was accepted by the government as adeguate to protect



the publiciand-used by Dow on its prOduct KURON weed and brush killer.J

"Labels of products in the third
category should carry the word "caution" in statements indicating the means of
avoiding the principal hazards of use. Use of the skull and cross bones, the
word IJ'poison? and antidote statements are not necessary for these products.T

Another part of the label which may need more explanation is the ingredient
statement. Ingredient statements?are required to be made on all pesticide products
and must be stated in certain ways. The constituents of a product must be
characterized as either active or inert ingredients. The active ingredients must
be described by their chemical name.

The two ways in which ingredient statements can be listed re specified

.Et-uua 7 mbt?ndAIKf ?f 5?
in section 362.7 of the-regulations. -These+were to list each active ingredient
and its percentage and the total of the inert ingredients or to list the total

of the active ingredients and each individual active ingredient in descending

order of percentage.without the actual percentage of each ingredient.plus the

16192

7hr W.- T.

 



a .- 1,4.
I - f?to. .-

n. 
.

1, 

.

total inert ingredients. Equal prominence is required to be giuen to all
of the ingredient statements and these ingredient statements were required to
. be placed on the front panel of pesticide products. In other words, only those
materials that were considered active ingredients from a pesticidal standpoint
were allowed to be listed in the ingredient statement even though?the product
contained other materials that comprised the total.

It has been suggested that should have been listed on the label of
this product.
standpoint since this is not an active pesticidal constituent; additionally, the
initials TCDD do not represent a common chemical name_acceptable to the federal
government so such listing would again be prohibited. The other reason for not
listing TCDD as an ingredient is that the limits of analytical detection at the
time of production of the KURON in question were such that TCDD was not found in
this product.

There are other sections of the label that are, of course, pertinent to
the allegatidns made concerning the product KURON weed and brush killer. Among
these are the instructions under a section titled TWarning" that "applications
by airplane, ground rigs, and hand dispensers should be carried out only when
there is no hazard from drift.? Other instructions which are meaningful to a
user of such products are the instructions that "coarse sprays are less likely
to drift" as well as the.instructions on application rates and times for specific
applications of the product.

The procedures followed for labeling pesticidal products are an application
of the principles that are included for labeling of chemicals set forth in the
American National Standard for Precautionary Labeling ANSI is a
consensus standard which was not published until 1976, howevercmdoes?hava the
benefit of review of such organizations as the American Society of Agricultural

Engineers, the National Safety Council, the American Society of Safety Engineers,

the Society of Toxicology, the American Conference of Governmental

 18193

.. .

Such a listing would not have been acceptable from the government?S'





'g'if 

I

Industrial Hygienists, and numerous other government, industry, and independent
- organizations with an interest in conveying precautionary information.
These principles prescribe a signal word, statement of hazard, precautionary

measures, and as appropriate, instructions in case of contact or exposure as



procedures for providing precautionary labeling. The labeling used by Dow on

KURCN weed and brush killer demonstrates these principles. - i
The procedures used within Dow to develop labeling in l967-involved the

initial labeling being developed from a use standpoint by a technical specialist

from our Agricultural Products Department along with the individual responsible

for submitting registration applications to the federal government, and

precautionary information coming from a labeling specialist like myself. This

draft copy then would be typed and sent for review by the numerous Specialists

within the Company with input on the health and safety aspects coming from

toxicologists, industrial hygienists, medical doctors, individuals from our

-1 5?

v3

5- quality function in connection with our manufacturing people providing input on?
the composition of the product, trademark and patent comments coming from our
attorneys, and the original three persons reviewing these inputs and those of

1..

our marketing personnel. Once these inputs had been composited, the labeling
was sent to the federal government for their acceptance or suggestions for Change
if they-did?not accept it.

Upon return of a copy of the accepted labeling, the labeling specialist
would initiate the actions responsible for getting the labels or the preprinted
container copy to our production people for application on containers of the
final product.

It is important to remember when developing or reviewing labeling for a
product that the labeling must be based on the product as a whole; consequently,

the labeling must be viewed in its entirety, not just in partse?since it is the

whole product that is used, not a part

I have compared the labeling used by other pesticide producers of Similar
products and find the-labeling used, which also was required to be Submitted
to the Pesticide Registration Division before being used, to be essentially the
same where applicable as that used on the product KURON weed and brush killer.
Based on my education, training, and experience, it is my opinion that the

label used on the product KURON weed and brush killer in the time period 1967'

_through 1969 met or exceeded all regulations and standards of care pertaining

to such products; and further the directions and precautions if followed were
adequate to prevent accident or injury with the handling and application of

the product.

99969? mm

is




 

 

 


W0 ms 

 

'AK-fr?

VOL 13. No. I
Fries-d USA.

Increased Susceptibility to Bacterial Infection as a Sequela of
Exposure to 

J. E. R. E. FAITH. E. E. KCCONNELL. urn J. A. 
National {mutate ?Environmental Health Sciences, Research triangle Pore. North Carolina 

Received for publication 12 August 1215

The e?'ects of subclinical levels of (TODD)
on the response of mice to infection with either Salmonella bent or Eerpasoirus
suie. also known as peeudorabiee virus. are reported. TCDD is a contaminant of
certain commercially use?zl chemicals. such as chlorinated phenols or herbi-
cides. It has been shown to cause thymic atrophy and to suppress cell-mediated
immunity in laboratory animals. Sublethal levels of TCDD were used: 0.5. l. 5.

signi?cant decrease in weight gain compared with control mice occurred at the



20-14 dosage. Dose schedules of 1 lug or more. followed by salmonella infection.
resulted in signi?cant increases in mortality and dun-eases in the time from
infection to death. However. TCDD had no signi?cant e?'ect on mortality in the
peeudorabies-ini'scted mice. The most important ?nding in this study is that
extremely low levels of TCDD. which do not produce clinical or pathological
change. still have the capacity to siren:- host defense. 

 

Various environmental chemicals have been
shown to afoot immune respons- m? but resist-
ance to infectious agents (Friend and Trainer
Gainer Gainer and P17 Hemphil] et
al. Jones et al. [141. Holler and ?l'hig'pen
Koller end Road: Keller Ver-
schuuren et al. [271. Voe and van Genderen
Vos et al. Voe and Moore Wesser-
mann et al. [321). These reports generally in-
volve observed defects in antibody response (14cell mediated immu-

nity (28. 29. 30). although several (8. l. 10. 11):
also describe suppressed host resistancetoini'ec- 

non.

(TODD)
can occur as a highly uni: consaminanrin the
production of some chlorinated phenols or chem-
icals from chlorinated phenols such
as the herbicide 
acid and thus could be unknownly disuibuted
in the environment. TCDD as an environmen-
tal contaminant can present health problems.
both in man and other animals. It is associated

with chloracne in man (16) and chick edema

disease in chickens (5). More recently TCDD
and 2.4.5-ttichlorophenol were implicated as
the probable cause of to:de in a group of
horses in eastern  in addition to the
horses. insects. birds. and rodents were poi-
soned. and there were several cases of human
illness attributed to TCDD in this incident (3).

has been observed to cause profound
thymic atrophy and suppress cell-mediated im-



unity in rats. mice. and guina pigs (23. 29). It
is not known whether TODD-exposed animals
are more susceptible to infectious agents. These
medias were d-igned to determine the e?'ects
of subclinical levels on the response of
mice subequently infected with either Salmo-
nella born or peeudorabies virus (PRV). The
criteria used to measure these effects were clini-
cal appearance. weight gains. mortality rates.
?rm from infection to death. and pathologin
examination.

MATERIALS AND METHODS

Animal. Four-week-old male 6573!?th (JCT)
mice were purchased tram
Jackson Laboratories. Bar Harbor. .Me. The mice
were caged individually under ?lter tops (Enviro-
gard. Lab Products. Gar?eld. NJ.) following a rigid
sanitary regimen and given food (Steril?aeble st-
Bios. Allied Mills. Inc. Chicago. HL) and water ad
libitum. 

TCDD. TCDD. greater than 99% purity (lot 351-
m-m. was a gilt from Dow Chemical Company.
Midland. Mich. it was dissolved in reagent-grade
acetone and subsequently diluted with at least 6
parts oi'corn oil. The exact volume of corn. oil was
adjusted to provide each mouse with a dose volume
ol? approximately 0.2 mi. 

infectious agents. The bacterium used was a S.
barn isolate from a naturally infected opossum. his
organism was grown overnight at 35 to 31C on
?l?ry'pticase soy agar containing 5% sheep blood. lac-
terial dilutions were prepared in sterile physiologi-
cal saline. and the number ofbecteria was quanti-

l" at?.

 

 



 

Ex. "8

12.15991.- M000

 

 
    

 



 



 

 

 

 

ll.

 

 

 



 




 

ll

 



 



"l

 

 

 

 

 

 

 

 

?il




 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

1320 THIGPEN ET 

tated toroidlnen-icelly vith a Coleman model 0120
neon-columnar? sea on In addition. the noon-
beroi'totel viable bacteria 
eoiotel inoculation wee con?rmed by conventional
total plate count More;

The viral mot need wee Home :eie (PIN)
(I). PRV tee produced in TS-cn? Falcon flesh coo-
teioine Eli-13 monolayers. After a Maine cytopetho-
[eoio 

cellular debt-la. The aoperoetaote from each fleet
were pooled. thoroughly mixed. ellquoted in 2-ml

Muddet-nCMMAtthetined.

inoculation the original mica contained ep-
proatimately 5.0 10' deeue culture jol'eotiee 
ml. The pooled viral anaemic: wee mime hege-
tire for otyooplaetoe end hostel-in.

etody. Lethal dole reepooee corvee
meetehll?edl?orSJern aodi'lvtodatemine

. challenge (0.2 ml incaoee-itooeelly 9.9.1) that

wold produce 20% morality in normal
mmdumebeervedforledeye. Deednice
were examined by both microbiological and bino-
petholoeicel proceduee to verify iu'ecn'ott

Experimental Two dilferent experimente
were performed. In eeoh experimeot- tweet-old
mice were rendotoly divided into tour group. Each
none 
week for i weeks. Hire were hood with

corn oil. 

let the ?rst experiment. tech received

daye el'tar the lee: ?ee when the mice
were I weeks old. tech group wee reodonly divided
loathe-economy. Hiainooe ?bump-ere
injected Lp. with 10' riehle 5. beta orptu?aooe
{Laundoeeh the eecood nbgroup received 10? TCID
dole). and the third subgroup 
troll wee injected with aalirue only. la the
mberot?hecterielorvirel aaeoterequiredtokill
approximately 20! of the once inoculated lithih 
li-dey obeemdoo period.) 

The second experiment e-ee conducted in toen-
mouse
received either 0. 0.5. 1. or 5 u; of per
week and the RV subgroup wee eliminated.

Both expel-imam tore perforated ee blind mid-
iee. and theorderd'ioocolatioo vee 



eninale inoculated no not etatieticellr dl?'erent
from that oheerved in the ?rst 259?- or the middle
50%. Therefore. the order of inoculation wee ur-
mally irrelevent to euheequent survival.

After hectarial end viral challenge. the animals
were obeereed et Mel daily for 14 daye. Body
engine were recorded et deeth or at termination of
the study. Net-romeo were performed on at laeot
three mice from each subgroup. end selected 
were obtained for hietooetholocical and microbiolog-
icel examination. Mice from lubfl'?m with Ice
than three deethe were eaorii?lced and oecropeied on
tier 14.

The eelected for hietopetholoeicel teemi-

 

 

Iit'retr. [Helm-t.

nation were ?xed in 101- oeotrel biza?ered formalin.

tinely stained with heuetoeyllo end 

The criterie deed to ampere the drone of tables
thel levele of TCDD in nice euheequeotly infected
with bent or PBV were clinical eweereooe.
weight gain. morality ratee. tithe from infection to
death. and pathological examination.

Statistical For the mortality data. dole
reepooee mode were analyzed by the northbou-
mm. 
I 2tehleet12). Foe-theether
varieblee. the alanine-ace oi' trud-
wee determined by the Joachheere tee: (13). and
treatmeotuconu-ol di??ereocee Iere evaluated by the
molded Sang-Whitney u. teeta (12).

RES ULTS

Eden: on weight gain. mortality
rate. and time from infection to death. There
wee e-entially no dl?'erence in mean poop
body weighte prior to TCDD doeing. By week 4.
the high-don group (20 pg of TCDD per kg)
gained eifnl?cently 0.01) lose weight than
did the coon-ole. The 10 per kcdoee
group eleo rained leea weight; all Other TCDD
my body Wei[l1tI"were comparable to the
group which received acetone-corn oil (Table I).
At a given level there were no signi?-
cant di?'erencea in the week 4 weight: among
the mice to the hectorie. virus. or
control mupe. There wee little be-
tween body weight at the time of infection end
euheequent Willi.

Mortality rates in the bacteria and Virus con-
trol groupe that had not received TCDD were as
entidpeted baled on the preliminary
mdy (Table 2). The [Taupe of mice expand TCDD per kg and sinus.
born tho-ed signi?cant (P 0.05 to 0.01)
inmortelityretee. Themortelityrete

Teen 1. .I-ooer

 

 

June exposure period 
Weekly
TCDD loin'el 
luck; as on [If 
duh-u 
Lent I
20 9 ll.? 0.1! 1.06 0.20'
10 5  ll.? 0.22 5.2! 0.5
5 I ?41:02 
0 an ll.? 0.19 0.17
hot 11
5 15 3.31 0.2! 4.56 0.17
l. 10 21.? 0.22 0.17
0 5 3.5 3.07 0.21 4.06 r. 0.19
0 13 21.13 0.21. L5 0.14

 




elen individualhodyvught been.

'Mean=eundarderror.

4' com.



?1698 M00

 

 

 

 

 

 

 

 

R.


u??enIU?I

  

e.

H.



Iiilil' ?l

ii!



 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 



V01. I. 1175

inch-0.5 mummpm

duct? law! hand on par-mu ofthil
study would biennium bar-run 0.5 and 1.0
par kg {Tabla 2; Fig. 1).

The morality 11m in tho groups that n-

Tnu 2. Wafl?daymm?tymmof

ch11me calm-5. 

Vlqu

 

 

 

 

mp warn-u!)
loan-1 Vin! an.
. 1
im?, tall
[ml
20 IIW 100.010) NW)
10 ?(13:20? 010201 ?all!
I I?m mm .1920)
0 mm mm um
but!
5 1011010? 
1 summ- ND ?11131
0.5 mm) ND 0101.!)
0 mm ND 0mm

u- indinhd H1 and? bun
'Md?hll??dth?luw



:mmuwuuamn.


"rho nub-I u: ?than Han In nah-rd
undying-rm? mailman-cup.

TCCD-DECREASED HOST RESISTANCE T0 INFECTION

 



Than was a signi?cant (P 0.01) dun-n-
hud don-um in tin: from infoaion tad-uh in
ms. barn-ch.th min (Tabla 3:11;. 1).

0c.
Em!

-..-

 
  
  
   
  
  
 
 
    

 

 

 

(MM humbly 1i.

 

If ..

Tm?

 

 

 


wanna. 
ND. Nu has. and l?buqundy with 3. km.
Tau 3. 
- Inna-ill Vin] 
I i; mu Na. d' Na. 
15" I?ll 
n. a? .W
$1911
20 19 2.35 0.4V 2 0.00 5.00
10 13 5.50, 0.6? 0 
5 13 1.54 0.04- 3 0.33 1.11
0 5 0.40 1.00 3 {.67 0.30
Exp: II -
5.0 14 2.00 0.3! 
1.0 13 5.03 0.32 . ?b
0.5 5 0.00 1.35 ND
0 5 7.40 1.81 -. ND

 

TCDD in lacuna-corn oil atriu- gun-1': incubation on u: individual body Insight bun.

Hutu-id ol'S. bu: via Lp. ingucn'on on day 2 uh: the wulldy duo 

Viz-db: tho In'?y can of TCDD. 
?Tho nnmhord'nommvon within 14 day: 11th arm-:1

cit-?ange.
i mndud 
0.01.
0.05. 2
ND. No: dam.

 

 

 

 

 

 

 

 

1321?
caved virus Inn not soon to diffu- signi?cantly .



 

 

 

 

 

 



 

 

 1 il'?

 

 

 

 

 

 

 

   
 
 
      
 
   
  
   
 
 
 
   
 
 
 
  
   
  
  
 
 
 
 
 
 
 

 

 

 

 

 

 

 

 



 

 





 

1322 TBIGPEN ET AL. .

Puholon'cnl ohoorvotlono. Pootmortom 
onu?notion ofmt'co trontod with TCDD only ro-
voolod lodono in tho livor Ind



?poup woo mollorthonnormol. 
commotion mulod cortiool otrophy duo to
o! omoll mophy woo

tumomuoondmnotoboo?odutbo
lot-or Intorootingly. no looiono . wot-o
found in tho opioon.

Tho Iivon from tho 20 Mpg-our: won
lightn- in color tbon oormol ond wot-o mottlod.
long/kgonimolo.
ondno wot-orbiblolntho
llvon h'ommloointho 5. tat-0.5 HITCDD
pot kg poupo. ?lo-topothobuicol ?omino-

voolodonodulorouxfuowith 
of cytomqoly ond 
fool ofoinglo coll nocrooio. A for in?nmmoton
oollo monocular) woo-o found
in tho ol' oounoio. In oddltion. thoro won
anon intraunulor accumulations o?in'dlnbop-
otocytoo. oopocinlly in mitonol moo. Laotian:

ornno.

Tho looiono doom'hod woro still opporont in
thomloo poupo inoc-
ulotod with vim or hot-toxin (both survivor:
ond honour-vim). In oddition. mioo in tho boo-
torlol mthotdiod tho-odmooro-undtnio-

 roocoplc loo-ions typical of oolmonollooio (1. 26).
Thu-o woo widooprood in?llntion of poly-m
loukocytoo. portimlorly in visual
organs. Foul nomtic hopotitio momblin; mi-
n-oioluonon woo ottribanod to odiocont 
of vucnlitio and thrombosis. nonto-
olo. thrombotic mlitio. ond ou'ppurodon
won also abound in tho oploon. Hound: ontor-
ids. ptn-nlont porltooldo. 1nd Ico-
purotivo Will the ootod.

In tho Vino portion of tho outpon'xhont. lo-
n'ono otm'hmbloto PRV mootoboorvod in
onyoftho mioo onminod. loborotory
rodonto io known to produco for loolono at tho
light low! (21).

DISCUSSION

Tho don in this study cloorly ootob-
liobod tho: mioo to four wooltly of
1.5.10. 
quontly lnl'ootod with on ofS. born
bnt'o hichor roto of mortality
with ohm-tonod incubotion poriod. Put-thor-
moro. to por
k; oppoorod clinically Ito-moi. bad 1 normal

Inna-r. [anoint


olorlool ovidoncool'toxioity. Ihlodomonotrotoo
tho mootimpomntupoctd'thjomdy. Lo"

that oxn'omoly low lovolo which do not 

wod'ttoo clinical or pothoth thong! sill


?pond to onv'ironmontol oontomhunto oath


myborondotodtnonmm'blotomin?


Tho an: that TCDD did not inn-om monol-

di?'oronoo in _tho potho'onooio d? two
ogonto. 


Thoro oro morn] hoot dol'onoo futon rhich
mud for tho incuood uncondhility to
focnltotivo organism such on S.
born. indudo: dofoc: in coll-mo-
diotod immunity oithor through lom of audio:-
ing oollo or by inhibition ofcoll function: (ii) 
dol?oct on tho human! oyotom; dol'oct in



Coll-modiotod immnno roosdom on ovi-
donood by doloyod hymmidvity hnvo boon
ohown to ploy on important rolo in immunity
ond from infoctiono with salmon-1L1
and othor focultotivo ?onions
(4. 15. 20. 22. 23. 25}. Doloyod byporoonn?tlvity
io thought to bo modlotod through woduodonof
by spod?onlly activatod toll: in
moon:- to bootm?ol ontigono (4. 20. 12. 23. 25].
TCDD boo boon shown to doloyod hy?
poroonoitlvity motion: ond rapt-non of? toll:
to ontivotion by tho mitogono oononnovolin A
ond phytohoqulutim?n (23. 29). Thou snadios
did not invoou'goto tho in?uonco of TCDD on
production. but it would bo surpris-

incil'TCDD did not oloo mppron tholr produo-.


T-coll function. .

onnt in impoirod hoot ruinonco to oolmonollo
lnl'ootion in mlco sinco full proton-ion from chol-
longo oppoon to dopond upon tho immuno ro-
tponoo that loodo to doloyId-typo hypononoitiv-
it'y. nthor to tho: which rooulto in drool?.
in: antibody and orthuo oonoitiv-icy H). Tho
o?'oot on humorol antibody production
in mloo i: not known. Recon: mdloo by ono of
tho outhnt's (R. E. Folth. unpublishod dotoJ
indium that TCDD hon no o?'oct on humorol
onn'hody production to bovino 3mm: [loban
in roto. In coma-not. tho gninoo. pig humor-o1
mponoo to totonuo touid ?u oboorvod to bo
impojrod (Von ot ol. [291). .

is; T698 MUG



 

Ill? 

 

 

 

 
  
 
 
 
 
 
 
 
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 



 

 

 

 

 

 

 

 

 

 

3. ?a .adnpun?gb mama. Egg 8 "2.160.502 

Earl-non- 9- Banana- 1. 3.34.8. F?nlitliluula?lli. 
HF. 2.9% 3.303.555: 2. guns. Ian-15.! pl 8 F. as.
man. as. Eli. Flag: in: E9 gmg?r??ig
uo?l?omEI-oa-zugouumgiua 
Eon 91 Earn. .1. .meIa on 
nul- nozu. 385 3? Ear-rad It. If: Ir
mama-.1 ?9520 
aorta: Econ 538.3..- uEHv?nun. 55.2.. Ens: ghg?lng
nun-5. uh and-non .8115 ERR 3:36917?pnillE?
Ilka.- il. ct. a.
.3515.

1.4 u. 2.5 Quill. ?55- 1. Hull. Kuhn-uni . u. . Fun-an- t. 
incl-?PtrnisgrnuEE-IL. Ina 
FINE lull: 5 I II. ml. Kl. 53.5.

I. Incl. g. 3.2.  .IBEI Faun-nil- .n
gringo gii?3.gn

. 35f:-


atria-1r $51555 s?gsg?n?u?ghgt
gr El. all. Mal?I8. 

u.  u. u. "Io. El Krill-Isa is U. Flu. h: KinkObi-n. n. D: u. g. L. ELF. a. n. anon-:1 ii?un lull-HIEI. at. ..
5n 1. n. gal. 53. MEG: E. Gnu. i pun. :27 
. . - 

a. Kuhn?rut. an. I. 0.. arcing018.1 E. mains? in n. all HIP H39 Eula-l ca ?Illallil..-
uni-sliding. l-nniuu i ilk?I 53313.5 on Slur-cu pl 53.8.3. haunt?Q1331 3.5 Fr . 
?oral" nus-Bl. In. moan winn- an. Pa  
all 5.0.3.13! u. L. 0.. .r Kali. i L. O. ?nE. 52. H. 

q. Elli. 0. K. Bill. In; a. o. run: "an. ?rfgilu on G. Elba 5. 

. .I .


 

 




 

 

 

 

 

 .Ii,//lii,

 

QLIBQE I 

I

 




   .  

 

 

 

 

 

 



 

 

 

 

 

 

 

 

 

 

 

 

 

   

 

 

   
 
 
 

     

 



 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

and (51 Other

Maf?arh.

Ddxn12019535
Shoecraft et al v. The Dow Chemical Company
LABORATORY TEST 
OF
- KAUKO SATAMA

BLOOD CHEMISTRIES
BLOOD COUNT ?bum?? 
Hca 15.2 (3) IGl?bulm 
HCT 45_0 Ratio LL1 (3)
NBC 23 or 04_1 Total Proteins 5-3 (3)
Alkaline 76 (3)
RBC .2224!) phosphatase 
I
Bilirubin 0.5 (3) 
MCH LDH 254_ (3)
MCHC 33.7 (3) SCOT 41 (3)
PTT .-. SGPT 43 (3)
C?ntr?l GGT (corp) 23 (3) I
7 
SR (Sad Rate, CO 2
2 3 
PRO. TIME I 14?6" I:
Control Sodium- (NA) 4 4 1
PL- 255 000 PotaSSium (K) 151 (3)
DIFF. Chloride (CL) 9 1 (3)
SECS C31Clum - 15 
I 3
35 (3) Triglycerides (3
i Mono 2 Creatinine 2; 
I Blood Urea 
- i- EDS Nitrogen
r' 3350- Uric Acid 4-5 (31
- Band T3 27.1 (3)
TESTS ORDERED BY T4 ll
PLAINTIFFS COUNSEL: Free Thyroxine 
199 Index 2.06 (3) I
Chol st 301 (3) 

193 HDL Cholesterol 37 (3) 
3.53 (3)
PLASMA EST 10L gg?al Cholesterol/
LDL Cholesterol 129 
- URINALYSIS: 

Co 10: we]. 10w VDRL 
Appearance 39;? VLDL
Specific Gravity i 013
J- 9? .14"
Protein -- 
Glucose --
I Ketonas GLUCOSE =35 -:
:3 09?91? gland ~11- PHOSPHORUS 3 . 5 
Bilirusin -- .
Casts/LP)? n; 'l
. 1-2 DIRECT BILIRUBIN 0.1
. . -  
  - . RECT BILIR BIN 0.4
- Epithelial Cells I or 
Bacteria a

Mucus 
(1) Gila County General Hospital; Laboratory;
tam

 

"nr4anq1 f?hnva?nwine-

16202

   DOW:  
I

w?w?-Wf:



n. COOK, M.D.. M.P.H.
(517) 636-1383

 

at

 

nadir}:- 


   

. .--

 .-    . . ..  .. 
REVIEW OF TRANSLATED ARTICLE: HARDELL, L. AND A.
MESENCHIME SOFT TISSUE TUMOUR-

 AND EXPOSE PHENGXY ACIDS 
NO. 40, 1978. (sic)' 

 

. -. - P's-uh

This was a confusing article to review: the Swedish to English trans?
lation was looseg'references were not included; the data was incomplete;
andp'there were multiple.inconsistencies or contradictions incorporated
into the body of the'article:

In this paper the authors addressed some interesting epidemiologic
problems: confounding; recall, selection, consistency, and causation.

I am not convinced they solved any of them satisfactorily. In addition,
the methodology they used, case-control.while rapid and flexible is prone
to greater bias than the more rigorous cohort approach. Although phenoxy
acids or chlorophenols may be associated with malignant soft
tissue tumors, this study does not prove causation. It simply points out
the need for "continued investigationsn"

Cases were drawn from patients hospitalized at the Dncologic Clinic
in Umea, Sweden, 1970?l977. Males with any malignant soft
tissue tumor {fibrosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma,
angiosarcoma, osteosarcoma, etc.) were included. Each malignancy has a
different_age distribution of occurrence. Unless cases and controls were



matched"within fairly tight_age ranges, the results were potentially can?



founded75:; gr"
The_selection factors leading to hospitalization at thisrclinipFwere

not discussed. .If foresters or pulp workers were more apt to he seen at

this center, the implications are obvious, the relative risks were

inflated.

 

IZEOAONH

Sausal 

16204

. .
- - . I .
 .12..-. - 

 

. . - uteri-gar -. . - -.

- 
- -lr - .

 

54. - 

I-vli??ir match;ng criteria were not adequately presented.

As a rule of

thumb, the reader should be able to replicate the investigation using the
article as a guide. I would be hard pressed to do that in this case. For
example, I don?t know what they mean by ?otherwise matching" on page 2.

In addition; they apparently used different matching criteria for live and
dead cases. They also chose 8 controls for each live case and 10 controls
for each dead case,' and then limited their analysis to.a ??to?l match.
hhy? They exc1uded suicides from the control group. Why? They made a



point of selecting controls who had worked up to 2.years prior to death. 
. 
Why? :55


be

C11

One matches to control confounding. Having matched in the design, one

has to match in the analysis. They did not. -


The statistical methods they quote are Miettinen?s. His methods do 





not have complete acceptance by biostatisticians and epidemiologists.
What do they mean on page 3, under Statistical Methods, by "effect of 
What percent participation did they have? They suggest 1002 among
cases and 99.232.among'controls. This degree of success is difficult to
believe. Unless other inyestigations by the same authors demonstrated
a high degree Of epidemiologic sophistication (which this analysis does
not). I would suspect a fair amount of selection bias has been incorporated
into this study.
-Since they had such poor environmental measurements, they were really
testing the'risk of being a wood product worker. The chemicals ?ipyv
arbitrarily choose as labels were merely surrogates for the wSEgee?? total

exposures. One has to ask, what were the workers other chemical exposures,

16205

  
 
  
     

 . . ?1.3m; 
-- . . -- hag Hui-3h: .10 3' 
 1?41In?. . - - . - 
1c?v?a "our I55: I I 
.1- 

phpsical exposures and



'selection'into or out of the jobs in question?

.-

Was there a dose response in any sense?

.. For me, the study did a better job of presenting questions than
answers. What is a Swedish municipality? What are the problems asso?

ciated with the register of population How did they use the

register of dead personS?

What was their definition 6f exposed? Was

it greater than "totally 1 day's length"? Why were "all persons in

the region with the studies tumor diseases not investigated at the
clinic In 602 of the cases and controls, eXposure histories

were obtained from a member of the "bereaVed family"; can your spouse

list your exposures of twenty years ago?'

R. R. Cook, M.D.
14 November 1978



f"
l? ?u

16205

 

emu DOW U.S.A. 2?7 7

 

MIDLAND. MICHIGAN 48640

November 10, 1976

 





W. B. Crummett - 574 E. S. Parsey 834 hi
B. Holder 607 B. A. Schwetz 1803 
a. L. Johnson 2040 w. w. Muelder - 9001 - 
M. J. Mintz 1710 w. a. Seward - 9001 3
R. D. Olson - 607 Jp

CHLORINATED DIOXIN ISSUE MANAGEMENT TEAM MEETING, November 17, 1976,
1:15 P.M., Planning Conference Room, Agricultural Products Department,
9008 Building. 

AGENDA

M. J. Mintz

Progress in the process for minimizing the TCDD in and
silVex products.

E. S. Parsex

Appropriate timing for announcing new TCDD product spec.

B. A. Schwetz

Review of NIEHS meeting.

B. B. Holder

Comments gathered from conversations with those

concerned with 
problems - Italy, etc. 

R. L. Johnson

Status of studies to determine how the chlorinated phenolic Dowicides
are used and their fate in the environment.

AN OPERATING UNIT OF THE DOW CHEMICAL COMPANY

0069191 M00 0

w. B. Crummett et al
Page two
November_10, 1976

R. D. Olson

a) Current status of employer monitoring study.
b) Status of soil monitoring studies for TCDD around and in the
Plant - including a protocol of the study.

W. B. Crummett

Current status of EPA Dioxin Implementation Plan.

w. w. Muelder

Status of investigations on occurrence of chlorinated dioxins
and furans in Dow products.

W. B. Seward, J. H. Davidson
Status of legislation and P.R. effort to support phenoxy herbicides.



1
Davidson I
Agricultural Products Department

nac

1069191 M00

18268

 

AGRICULTURAL EXPERIMENT STATIONS . 
UNIVERSITY OF FLORIDA 
EVERGLADES EXPERIMENT STATION

December 23, 1961+ .. 0. .

IILLI OLADI. FLOHHDA

3:430

Anna

Dr. Lawrence Southwick
Agricultural Chemical Development
The Dow Chemical Caspany

Abbott Road Buildings

Midland, Michigan

Dear Larry:

The following comments are in response to Chuck Lichy?s phone request
concerning silvex (Kuron) for sugarcane in Florida.



Information on the performance of silvex under our conditions has been
sent Dow on several occasions in the form of mimeo reports, data elmaries and
letters. Continued, long-time use of 2 ,h-D in parts of our cane area has per-
mitted the development of broadleai? weed populations resistant to Prop-
erly timed. applications may control some, but not all, of these species
effectively. These to which I refer are dogfennel, ground cherry, 
purslane, ragweed. and wild. lettuce. Other low-growing, less competitive winter
annuals which are less important in cane culture may not be controlled by 
and/or II?he weeds which I listed are amenable to control-with silvex.

I think that silvex is a. needed and useful tool in case culture which I
would recommend if it were cleared. My use suggestions would be for postemer-
gence application of 1 - 1% lb./A a1 (and not over 2 lb. in extreme cases)
silvex to the species named after establishment of plant cane or ratooning of
stubble cane up until the cane is closed-in. I would stress that the rate of
application should be adjusted to weed maturity.

Please contact me if you wish further information.
Sincerely yours,
. /7
Gregg-?xe-
1, 
J. OR?iNlco
Assgciate Horticulturist

JRO 

18203





2821

Reprinted from BioScr mm?

(5 The Public Health Implications of Widespread

Use of the Phenoxy Herbicides and Picloram

This paper is concerned primarily
with 2.4-0, silvex, and
ptcloram?the phenoxies because of
widespread use; picloram, and
2.4-0 more specifically because of their
use as military defoliants. The purpose
or this di5cussion is to explore possible
public health consequenCes of wide-
spread Vuse. This presentation will be
confined primarily to information as it

TABLE 1. Tolerances proposed to FDA

Julius E. Johnson

Table I shows a condensed version of
tolerances proposed for 
MCPA, silvex, and picloram. Those
marked with an asterisk are for toler-
ances at a permissible level for in
rice and flax seed and for picloram in
forage grasses and meat. ..The remainder
are negligible residue tolerances.

Table 2 demonstrates the infre-
quency of residues of phenoxy herbi-

 

 

2.4-D MCPA Silvex Piclorem

Fruits 
Berries 
Grains 

Rice 

Flax Seed 
Potatoes 
Sugarcane 
Hay 
Forage Grasses x?
Meet x-
Milk 

 

(All negligible residue tolerances except those marked'l

may translate to the health and safety
of man. The data presented will empha-
size exposure. toxicity, and chemistry.

First, i will comment on exposure to
food. wzzer, and air. There is very little
exposure of man to the phenox'y herbi-
crdes through food. The principal uses
of these herbicides in growing food
crops were originally registered on a
no-residUe basis. A change in policy now
requires these registrations be supported
by a negligible residue tolerance. An
industry task force has formally re-
quested FDA to establish these toler-
ances. (The request for has been
temporarily withdrawn without preju-
dice to future filing.)

 

The author is Vice-President and Director of
Research. Dow Chemical Company.

Presented II the Symposium on Possible
Public Health implications of Widespread Use
of HerbiCides (Chairman: Dr. Arthur Galston)
American institute of Biological Scrences,
Bloomington. indium. 26 August 1970.

September 1,1971

cities in foods as confirmed by the FDA
market basket survey of pesticides in
composites of 12 categories of food-
stuffs. Of the 1548 samples analyzed,
only 36 positive identifications were
made. All but one was less than 0.2
ppm. was found in three of the
surveyed samples (Duggan et 31.. [967;

Martin and Duggan, l968;Corneliussen.
I969).

Air and water are potential sources
of exposure of man to these herbicides.
Manigold and Schuize (1969) have re-
ported on pesticide residues in selected
streams in the West such as the Missouri,
Yellowstone, Colorado. Arkansas, Rio
Grande, Snake, and others. Phenoxy
herbicides were detected as shown in
Table 3. Of the 320 samples taken,
fractional parts per billion concentra-
tions were reported in 78 samples. The
measurement of picloram residues in
water has been reported following direct
spraying of a static pond in Texas
(Hoffman et al., 1969) and a watershed
experiment was conducted irt North
Carolina using and pic-
loram (Sheets and Lutz, l969). The
results of these studies are summarized
in Table {hit is noted that picloram
persisted longer than 2.4-0 or 
Air samples collected in spring and
summer from two wheat-growing areas
in the State of Washington Were ana-
lyzed for phenoxy herbicides by Bames-
berger and Adams (1966). The results
are summarized in Figure l. Phenoxy
herbicides are widely used for early
postemergence control of weeds in these
craps. There were detectable levels of
2.4-0 and 2.4.5-T in the air up to 0.06
rig/m3. Man will inhale about 30 in?5 of
air per day. At a level of 0.06 lug/m3,
this would amount to an exposure of

TABLE 2. FDA market basket survey pesticide residues 1965-68

 

Number of Samples with Residues of 1548 Samples Analyzed

 

PPM Total

0.001 - (0.1 0.1 - (0.2 0.4
2.4-0 23 4 27
MCPA 2 1 1 4
2.4.5-1' 2 0 3
Sin: 2 0 0 2

 

?sticide Monitoring Journal, 1967, 1968. 1969

Duggen, n. 5., H. C. Berry, and. L. Y. Johnson. 1967. Ferric. Mona. J. ml: 2-12.

Martin, Fl. J..lnd R. E. Duggen. 1963. Ferric. Morris. J. 11-20.
Corneliussen, P. E. 1969. Peeric. Mom?r. J. 140-152.

10210

899

ANOG

(J

8'7

rphenoxy herbicide per day per

man. For a "IO-kgman. this would be

0.025 as per kilogram body weight per
day of exposure.

A. variety of pesticides were reported
in dust trapped in rain collected in
Cincinnati in [965 (Weibel et al., 1966).
The dust was created by high winds over
the southwestern United States and
moved in a cloud in a northeasterly
direction to Ohio where the precipita-
tion occurred. An amount of 0.04 
of was determined to be present
based on the air dried weight of the dust
in the rain. No mine was given for the
amount of dust in the water. if it were
as much as lOS?e, the water could have
contained a 4 of The
ngnificancc of these findings are open
to question. hawever, because oi the 
difficulties in analyzing for such small
amounts, and (2) the alleged presence of
a newer pesticrde, ronnel, which is used
in relatively small amounts as an addi-
tive to cattle feed and as a topical spray
on cattle. The results are summarized in
Table 5

Direcr skin contact together with
vapor and mist exposure are encoun-
tered in varying degrees by applicators
and employees in manufacturing plants.
The exposure of an infrequunt user is

tuch less than professional pest control
operators or those engaged in manufac-
turing 5 days a week. A medical evalua-
tion of the effect of and 
has been made of men engaged in their
manufacture. The workmen were given
extensrve examinations includ-
ing a battery of at least 20 laboratory
tests. The results of these studies are
given in Tables 6 and No meaningful
differences were noted when the clinical
results on these workers were compared
to those obtained on a control popula-
tion of 4600 men not exposed to 2.4-0
or An important observation
was that there were no chromosomal
effects.l

in the fall of I969, considerable
apprehension was created in the minds
of the public when an announcement
was made of tests Sponsored by the
National Cancer lnsritute reporting that
2.4.5-T was teratogenic. Following that
announcement. i made special efforts to
trace the identity ol the samples used
and reported to representatives of the
National Cancer Institute and the Mrak
Commissron that the test sample proba-

contained 2.4.7.8-tetrachlorodi-

 

lUnpublished dala. I970, Dow Chemical Co.

900

TABLE 3. Residues in water from 20
monitoring site: no 16 mtern states 1965-68

 

 

No. Pasitivie
per Parts per
320 Sample: Billion
2.4-0 36 0.01 0.36
28 0.01 - 0.0?
Silver: 14 0.01 0.21

 

(Pesticide Monitoring Journal, MInigold and
Schulzo, 1969}

benzo-p?dioxin in amounts which might
have been a complicating factor. The
sample employed in the study was
analyzed and found to contain about 27
of 
dioxin. Teratology tests with 
and the impurity were conducted in
Dow laboratories. Results of the ?rst
experiments were observed by represen-
tatives of the Department of Health.

Commercial containing less
than 1 of 2,3,7,8-tetrachlorodi-
benzo?p?dioxin was administered by
savage to groups of pregnant female rats
on days 6 through IS of the gestation
period. The doses of adminis-
tered were 0 (controlmg/kglday. No clinical
pathological signs of adverse chemical
effects were observed in treated dams
during the period of treatment or gesta-
tion. Fetuses were harvested by cesarean.
section on clay 20 of gestation. Visual
and histopathologicsl examination of
the fetuses failed to reveal any terato-
genie or embryotoxic effects (Emerson
et al., 1970).

In another test, commercial produc-
tion gade given to rats at a
dose of 50 mg/kg/day from days 6-l 5 of
pregnancy, produced a minimal amount
of fetotoxicity with one fetus in 203

 

 

Education Welfare. having intestinal hemorrhage. When
TABLE 4. Residues in water
. . Days
Type of Treatment Herbicide Lia/Acre PPS After
Water
.. Spray
Direct spray on pond Pictoram It 2400 (1
700 ?l
150 23
6 180
Spray on 25% of watershed 2.4-D 4 1800 - Pictoram 4 4200 2 I
340 21

 

Following is a description of the
series of experiments concerned with
the teratology question.

 

 



PULLIIMI 2' a
laSn

2.4.54
9

. 

WASH 

1V6 HICROGRIUS PER CUBIC If if?

at:
VAPOR

Fig. I. Phenoxy herbicides in air (?anne-
berger and Adams. I966).

commercial grade was given at
100 mg/kg/day, a high incidence of
maternal deaths occurred, and of the
pregnant animals that survived. most
had complete early resorptions. No fatal
anomalies were observed as a result of
the administration of the commercial
production grade to rats. Like-
wise. no abnormalities were noted in
litters of treated dams which were
allowed to deliver and raise their young
through lactation. Treatment had little
effect on the survival of pups through
this time period:2

Rabbits (New Zealand White) were
used in another experiment to deter-
mine the embryotoxicity or teratogen-
icity of commercial containing
less than 1 of 

 

2Unpublished em. 1970. Dow co.

BioSciencc Vol. 21 No. I7

or gross -

 

18211

 0.04

 

benzo-p-dioxin. Groups of pregnant
female rabbits were given the compound
by gavage from day 6 through 13 of
gestsiion. The doses of 2.4.S-T adminis-
tered were 0 (control). 10. 20. and 40
mg'kg/dav. Fetuses were harvested by
cesarean section performed on day 29.
No clinical or gross pathological signs or
adverse chemical effects were observed
during the period of treatment or gesta-
tion. Examination of the fetuses showed
no dose-related teratogenic ur embryo?
toxic eftecr (Emerson et al.. 1970).

in order to check the effect of the
contaminant itself. a grOup of pregnant
were administered 2.3.7.8-tetra-
cldorocil?enzo-p-dioxin by gavage on

days 6 through 15 of gestation. The-

dosages used were 0 (control). 0.03,
O.l25. 0.5, 2.0 and 8.0 pg/kg/body
weight/day. The fetuses were harvested
on day 20 of gestation. No differences
were observed in the fetuses taken from
dams treated at the dosage of 0.03
rig/kgjday and those taken from dams
that received the solvent vehicle only.
At the 0.125 pg/kg/day dose level. there
was evidence of intestinal hemorrhage
and subcutaneous edema in some of the
fetuses. At the 0.5 and 2.0 ug/kg/day
doses. there were a substantial number
ol fetal deaths. intestinal hemorrhage in
the fetuses was frequent. The 8
pg/kg/day dosage was toxic to the dams.
There were no Viable fetuses U1 the dams
which were examined on day 20 of
gestation. All resorptions occurred early
and no evidence of fetal tissue was
found. Skeletal examinations revealed
delayed OsSiFication of some sternebra
and skull bones. This occurred generally
throughout the venous poups. includ-
ing controls. and was not considered to
be of practical significance. The results
of this experiment indicated a high level
of maternal and fetal toxicity to be
associated with 2.3.7.8-tetrachlorodi-
benzo-p-dioxin. its presence in 
at levels of 27:8 could well have
accounted for the observations reported
and attributed to in the original
study conducted by the Bionetics
Laboratories (Sparschu et al.. 1970).

September 1. 1?71

2.4.5-T. and l2) pure 2.4.S-T to which
Specific amounts of the 2,3 .7.8-tetra-
cidurodibenzo-p-dioxin were added. The
results of these studies show that a
dose of 50 of from
day 6 15 of pregnancy
(maximum tolerated dose for our strain
of rats) did not cause teratogemc effects
in the youna. and t2) that when a toxic

.l?tll?

- dose of the 2.3.7.8-tetrachlorodibenzo:

p-dioxm was superimposed upon this
dose or 2.4.5-T. cleft palates were
observed in a few fetuses in addition to

solvent. Although the purr 2.4.5-T was
reported to have shown teraIOgenicny.
dosages of 100 mg/kg were required it
produce the cleft palates oi kidnex
involvement.4 

Steinl'elds also reported tertila :r
hamsters injected with commerua-
2.4.5-T containing 

tetrachlorodibenzo-p-dioxin at a dose 0' 

100 Terata were am- oh
Served in hamsters injected with EU mg
of 2.4.5-T containing 45 0! the
tetrachlorodibenzo-p-dioxin per kgldav

TABLE 6. InOJllrIBl health experience?manufacture of 2.4-0 the Dow Chemical Company

 



I
Work Experience

Range of Exposure

Results

 

No. Men Exposed

220 0.5-22 years

10 men exposed to 2.4-0 were karvotvped.

No dillerencoa whet
compared to :?nntro
population of 4600 mm

30-40 mglda

Result. No ellect on structural integrity or rearrangement of the genetic material of the

 

chromosomes.

the typical toxic manifestations of the

when given air-{2C However, it is very
important note that when no effect
levels of 
dioxin and 50 mg of pure 2.4.5-T were
combined. nu trratogenic and only
minimal results were ob-
served.J The rexairs are summarized in
Table is.

3Unpublished data. IQTO. 00w Chemical Co.

Verrett has reported that 2.4-D
2.4.5-T and silvex produce terata and
chick edema following injec
tion into the yolk sac. The extreme

4Testimony before the U5. Senate Commit
tee on Commerce. Sub-Committee on Energy
Water. Natural Resources and the Environ-
ment. l5 April 1970. Dr. Jesse Sleinfeld
page: 167-1159. Report of the Committee
Serial 91-60 (I970).

5Testimony before the US. Senate L'ummir
tee on Commerce. Sub-Committee on Energy
Water. Natural Resources. and the Environ
merit. l8 lune l9?70. Dr. lease Stemfeld

TABLE 7. Industrial healih experience?rnanuflcture of the Dow Chemical Company

Number of Dav: Exposed to

 

28 mg 2-4751. Dal? Number of Men Examined Results
1 59 24 No differences when
60 119 15 compared to control
120 - 239 5 population of 4600 men
240 - 479 men exposed to were karvotvpod.

FIE-suit: No offset on structural integrity or rearrangement of the genetic material at the

chromosomes.

901

its

0i?6uTZ. 

212

 DOW 712341 

TABLE 9. Teratologv study of with sheep

sensitivity of this test procedure makes
it difficult to interpret the meaning of
the re5ults from a public health point of
view.?

ans of Utah has tested the effects
of exposures on the l4th to
36th day of gestation of sheep. the
penod in which they are most suscep-
tible to teratogenic effects from the
plant 'erarrum californicum". No
anomalies were noted as a result of the
exposure to 2.4.5-T (see Table 

A study to determine the toler-
ance 01 pregnant female rats to silvex in
preparation to conducting a conven-
tional teratology study in rats has been
Silvex was fed to groups of
pregnant rats (5 to 6 per group) by
gas-age on days 6 through 15 of the
gestation nenod. Doses given were 100,
'50. 200. and 300 
wer no ettecls on either the dams or
lctmes harvested on day 20 by cesarean
motion at the dose level of 100
At 150 mg and abOVe there
was toxicity noted in the dams with an
met. 3th amount of maternal mortality
With in:reasing dosage. No- teratology
was ol?Sen'ed by grtiss examination in
fetuSes at these levels; however, there
 \omc resorpticns.3

~l c: ?tology experiments have also
"ecn conducted with 2.4-D. To date we

ave concluded studies on pregnant rats
writ. 2.4-0 acid and with propyl-
ene glycol butyl ether ester. In this
study. involving about 320 rat litters.
we administered corn oil as a control.
the acid form of at dosage levels
of 12.5.2150. 75. and 87.5 mg/kg/day

?Trwmony before the U5. Senate Commit-
tee nn t?um merce. Sub-Committee on Energy,
Water. Natural Resources. and the Environ-
ment. lti June 1970. Dr. Jacqueline Verrett,
pages 190-203, Report of the Committee.
Serial 91-60 (1970).

711mm. W. 1970. Poisonous Plant Research
thuratury. U.S.D.A.. Logan. Utah. Unpub-
lished dutn.

data. [970. Dow Chemical Co.

 

Amount No. of

Number

 

compound Fed Ewes Pregnant Ram?

None None 20 15 15 ms bore live, normal
lambs. 1 twin.
1 ewe aborted ?28 days,
fetus normal.

100 11 10 10 em bore live, normal

Dow Prod?n. lei lambs.

PGBE ester of 100 min/kg 11 9 7 ewes bore live, normal

lambs, 1 twin.

Dow Prod'n. 1 ewe died 36 days,

aapirared feed. Fetus 
normal.

1 ?ea died 35 days, profuse
diarrhea from 7th day.
Fetus normal.

 

W. Binne, Poisonous Plant Research Laboratory. USDA. Loan. Utah
llCompound given by stomach with 0.5 to. ground alfalfa. Dosod 14-35 days of

gestation.
ntetacarpels, haretip and cleft palate.

and the ester form at dosage levels of
'20, 40, 81. 122. and 142 mg/kg/day.
About one-half of the litters were taken
by cesarean section on day 20 of gesta-
tion for examination for soft tissue and
skeletal abnormalities and the other half
were allowed to go to term. The off-
spring were obServed for 21 days, after
which they were sacrificed and exam-
ined. Fetal weights were reduced at a
dose of 50 mg of 2.4-D acid/kg/day. and
at a dose of the ester at 121 mg/kg/day.
No teratogenic effects were noted.
Among litters delivered naturally, treat-
ment with during pregnancy had
no effect on neonatal growth and sur-
vival.9 .

Groups of male and female rats were
maintained on diets containing 0, 3000.
1000, or 300 picloram through a
three-generation reproduction study
without evidence of adverse effect as
Judged by indexes of fertility. gestation.

 

9Unpublished data. I970. Dow Chemical Co.

TABLE 8. Teratotoqy study in rats 
added to 50 mg pure 2.4.5-Tfkg body wild-av

 

 

No. of No. of Litters with Litters with
Treatment 
Litters Fetuses Cleft Pazate Hemorr.
Control (vehiclepure 2.4.5-+0.01 ug TCOBD 15 150 0
+0.03 pg 17 173 0 0
+0.06 ug 155 1 3
*?125 902

Vererrum caiifornicum fed this period induces cyclopia, short metatarsals and

Viability. lactation. by body weight, and
by teratological examination of fetuses.lo

A pilot study has been concluded to
determine the tolerances of pregnant
female rats to picloram in preparation
for a conventional teratology study.
Piclorarn was administered to groups of
pregnant rats (5 to 6 per group) by
gavage on days 6 through [5 of the
gestation period. The doses.given were
250, 500. 750. I000. and 2000
mg/kg/day. There was no adverse effuot
on the dams. nor to fetuses taken at day
20 of gestation at the levels of 250, 500.
and 750 At the [000
mg/kg/day dosage. there was no adverse
effect on the dams, but there was a
small increase in the absorptions. The
fetuses harvested showed no trrato-
genesis. At the 2000 mg/kg/day level.
only one dam survived to the 20th day
of gestation. Eleven fetuses were har-
vested from this dam all of which were
viable and appeared normal by gross
observation.lo

Since October 1969. great interest
has been created in 2.3.7.3-tetrachloro-
dibenzo-p?dioxin as a possible serious
and continuing contaminant in the
environment. It is important to empha-
size that the tetrachlorodibenzo-p-
dioxin is no longer a significant contam?
inant and probably never was in
carefully prepared commercial 
When it did occur?in 1964 and
1965?the contaminant carried over
from the intermediate 

 

loUnpublished data, 1970. Dow Chemical
Cu.

BioScience Vol. 21 No. l7

6]

18213



was established to ensure that no more
than minimal amounts would occur in
the product (the best method available

had a sensitivity of 1 ppm). improved

assay methods now show the actual

contaminant is less than 0.5 ppm.


Cl  10M Cl Cl HEM,

Cl Ky Cl ~00

Fig. 2.

tumult 
sun Sett?u'"
Mg. Lg ?Gin. u-i 

:1 ."t9 l?aotlu?!

Fig. 3. Effect of radiation from GE. sunlarnp
at distance of one meter on 12 Iolution
of 2,3,7 

Incidentally. the 2,4,5-trichlorophenol
submitted by the National Cancer Insti-
tute in 1964 to be tested by the
Bionetics Laboratory did not cause a

teratulogical response. This was a
sample ptOCured from  Coleman
Mathcison Bell who got it from

McKesson Robbins who got it from
Dow.

The following information is known
about the stability and prOperties of the

in solution it is rapidly degraded by
ultraviolet light at that
appear in the spectrum of the sun (Fig.
the tetra dimtin has onefifth the
solubility of DDT in water and one-
sixteen hundredth the solubility of DDT
in benzene (Table 10). Thus there is
probably less tendency to concentrate
in fat. but the question is really aca-
demic because the quantities of tetra

September I, 1971

benzene 0.047 91100 ml 78 91100 ml

 

dioxin in the environment are exceed-
ingly small.

Concern has been expressad that the
combustion of (even though the

Cl 0 
C) 2 NoCl
PRESSURE Cl. 0 :1

parent material contained no tetra-
chlorodibenzo-p-dioxin-) would cause
dioxin formation. A laboratory experi-
ment was conducted wherein agent
Orange (a defoliant containing equal
quantities of the n-butyl esters of 
and was applied to 13.5-cm
?lter paper at rate of 10.9 kg per 0.41
hectares. This is equal to approximately
4.5 kg of acid equivalent per
0.41 hectares. The paper was burned
and combustion products plus air drawn
through at: absorber packed with glass
beads cooled with liquid nitrogen. The
combustion products plus the ash was
solvent extracted and analyzed by gas
chromatography (Fig. 4). The burning
temperatu:es encountered were as

'follows:


600 - 800 300 - 400
1000 -1200 550 - 650
1300 - I400 700 - 750

There was no 2.3.7.8-tetrachlorodi-
benzo-p-dioxin detected in the combus-
tion products at a sensitivity of 5 ppm.
A similar experiment was tried using
wood as a substrate, but there were
substances in the combustion products
which interfered with the analysis.
Speculative claims are widespread that
resulues on vegetation might be
converted to 2.3.7.8-tetrachlorodiben-
zo-p-dioxin if the dead foliage is burned.
All available evidence to date indicates
this conversion dOes not occur. The
alleged precursor is in dilute form on
the substrate. the reaction is biomolecu-
molecules must be forced close
together to react.

Concern has also been eXpressed that
traces of preformed dioxin contami-
nating 2.4.5-T sprayed into the envtron-
men: would accumulate. The evidence

Paper surface
Body of flame
Front of flame

. 

- .. ..-  
24 (75 F). This evidence of degrada-
tion is so slow it will require flirther
observations to quantity.1 1

2 . 3 7 8-tetrachlorodibenzo-p-dioxin
does not move in soil. It is strongly
adsorbed on clay and organic matter
and is practically insoluble in water (0.2
parts per billion). First studies on up-
take by plants indicate none from
water and very little. if any. from
soil.12

(3
C)
12

TABLE 11. Solubilities
@000 ml of solution at 25 Cl

 

 

 

Solvent Pictorarn -
Potassium Salt
?3
Water 0.043 40.00 
Benzene 0.02 b3
Xylene 0.01 to
Ti

 

DOW 19??
Fig. 4.

Silvex is also made with 
chlorophcnol as a starting material. A
total of eight lots of silvex has been
from l967. 19ml.
and I969 production. No detectable
amounts of 2,3,7,3-tetrachlorodibenzo-
p-dioxin have been found using a
method having a sensitivity of 0.1 ppm.
Before leaving the subject of contami-
nating chlorodibenzo-p-dioxins. it is
noteworthy that no detectable dibenzo-
p?dioxins (including the 2.3.7.8-tetra-
chloro- or the 2.7-dichloro-) have been
found in samples analyzed. The
sensitivity of the analytical method used
was 1 ppm.13

 

?Unpublished data. 1910. Dow Chemical
Co.

Izl?rivate communication from Dr. P. C.
Kearney, 1910. USDA. Beltsville, Md.
I"Unpublished data. l970. Dow Chemical
Co.

903

182i

4

 712943

TABLE 12. Acute orIl toxicity, lingo out

 

L050 as Milliylms per kg Body Woimt

 

 

Rat Guinea Pig
2.4.0 375.0 ass.? i000.0
500.0 380.0
Siin 5ND 350.0
Piclonm 82010 BOND
DDT 150 - 300.0? 400.0
Parothion 1.7 - 30.0' 93-320?
0.022 - 0.045? 0.0005
aVaries according to vehicle or sex.
The formation of 2,7-dichlorodi- over, 2,7-dichlorodibenzo-p-dioxin is

benzo-p-dioxin does not occur in the
manufacture of 2,4-dichlorophenol
since it is made by direct chlorination of
phenol and not by alkaline hydrolysis of
a trichlorobenzcne. Reaction conditions
do not favor biomolecular condensation
to form a chlorodibenzodioxin. More-

TABLE 13. bio-effect levels 

 

 

Species 2.4-0 Silver: Piclorsrn
Reproduction and Fertility
Rat 25 150
2~Year Dietary Feeding 
Rat 62 5 150
Dog 12 5 350

more dif?th to form than the

even when reaction conditions are favor-
able.

The persistence of pesticides in soil
and water is an important factor  their

TABLE 14. Grass production?Panama

potential to occur as residues in foods
and water or to come in contact with
nontarget organisms. Degradation of
herbicides is accelerated by increased
temperature, soil microflora, and or-
ganic matter. A comparison of degrada-
tion rates in soils has been published by
Kearney et al. (see Fig. 5) (Gunther,
1969). it is true that pictorarii is more
persistent. Unlike DDT, however,
picloram forms salts that are water
soluble. The solubilities are shown in.
Table l. Picloram does not concentrate
in fat. Moreover. the- mammalian tox-
icity of picloram when compared with
most other herbicides is low. The po-
tential hazards from picloram will result
primarily from effects on sensitive plant
species. Up to the present time the
bioassay is more sensitive than chemiCal

 

Tons of VewtationlAcre 15 Months Deferred Grazing

 

Treatment Joraguo Grass Other Grasses Live Brush Taial
None 1.80 0.28 12.65 14.43
0.6 lb. pictorarri 
2.4 lb. 2.4-0
Per acre 13.55 0.29 0.50 14.34

 

Suzey and Montana, 1968 (26l

 

 

 

Persistence of herbicides in soils. Length of bar repleuentl time required for 7.5 to l00%

ion of the compound (after Kearney. 1969).

904




MCPA
2,4MONTHS

techniques for determining the presence
of picloram.

The acute oral toxicities of 2.4?0.
silvex. and pictoram are given in
Table l2, in comparison with DDT.
parathion, and the 
benzo?p?dioxin. It is evident that this
dioxin is highly toxic. in fact one of the
most [one substances known. This
alorie is a good and sufficient reason to
keep this impurity at an absolute
minimum. By contrast. the low order of
acute toxicin of picloram is alsr- of
interest. The no?effect levels attained in
long-term dietary feeding studies also
indicate a low order of toxicity of
pit-loram. Concentrations were not
tested above mg/kgiday (see Table
Ill. The reproduction and fertility tests
were conducted through three genera-
tious of rats receiving continuous
feeding of the test compounds. No birth
anomalies or other adverse effects were
observed with continuous intakes of 25
mg/kg/day of or with 150
mg/kg/day of pictoram. respectively.

Some of the benefits of using herbi-
Cides are illustrated in Tables l4 and IS.
The resulting incrcaScs in beef produc-
tion are important to nutrition and

bioScienCe Vol. 2] No. 17





16215


'1

ti

-3.

TABLE 15. Grass production?Oklahoma

 

Application

Grass. Lb./Acre

 

 

PM Ac" Weed Months Deferred Grazing w" Ya? Dry Ya."
None out. 6 862 3-48
2 "1-2.4 Oaks 5 2487 2200
3 Oaks 6 2265 2063
Oaks 6 2917 2083
4 Oaks 6 - 2750

 

4 

 

Elwell, 1964 (27?

health of man (Gunther. Swezey
and Montano. 1968).

In conclusion. I set forth the opinion
that the widespread use of phenoxy
herbicides has produced no demonstra-
ble cwdence of potential harm to man.
The herbicides used most widely (2.4-0
and 2.4.5-T) are degraded and do not
bmconCentrate. MoreOVer. the compare-
tive toxicity shows these materials to be
tolerated in a variety of test sys-
tems. Man is not exposed to harmful
Concentrations. Impurities can be an

important {actor?particularly the
chlorodibenzo-p-dioxins?but these can
be controiled by proper manufacturing
techniques. As always, care in applica-
tion is an important part of safe practice
both from the standpoint of man and
the ecosystem in which these tools are
used.

References
Bamesberger. W. and D. R. Adams. 1966.
Organic pestundes in the envu-onment.
Advan. Chem Ber

Corneliuuen, P. E. I969. Pem'c. Monir. 
140-152.

Dumn. R. E.. H. C. Barry. and L. Y.
Johnson. 1967.Pern'c. Moni't. J., 2-12.

Elwell. H. M. 1964. Apart. J.. 56:411-415.

Emerson. J. L.. D. J. Thompson. R. 
Strebing. C. G. Gerbig. and Robinson. 1970.
Manuscript aibmitted for publication to
Food and Cosmetic: Toxicologi'.

Gunther, F. 1969. Residue Reviews. 29,
Springer-Verlag. New York.

Hoffman, G. 0., E. D. Robison. and M. G.
Merkle. l969 WSSA ABST. 75. Las Vegas.
Feb.

Manigold. D. B.. and J. A. Schulze. I969
Punt. Mom'r. 1.. 3: 124-135.

Martin. R. J. and R. E. Duggan. I968.Pesn'c
Monti. 1.. ll-20.

Sheets. 1.. and V. F. Lutz. 1969. Bull N.
C. Exp. Sta. 

Spuschu. G. L.. F. L. Dunn. and V. K. Rowe.
1970. Manuscript submitted for publication
to Food and Cosmetics Toxicology.

SWezey. A. W.. and A. Montana. 1968. Down
to Earth. Summer. p. 6-9.

Weibel. S. R.. R. B. Weidner. l. M. Cohen. and
A. G. Christianson. i966. J. Amer Water
WorksAssoc. 58: [0754084.

c:
0


treat;

18218

2832

  
 

        

        
   

    
   

magi?  . ..  t: . mugs-Fri:-
r" ?7??Jm ?1 bh'J . . 533-" 5"4' 

w-  --. Ewell-J- 
44? 1? Lin- 4 c: --. +1 :Lm-i-z a Trad-- aux-'- -. 
- La -- L15 q'r AI- nadng?gaarr-sr; 'rhmanner-3.19.7.2: EmittinCBC 084770191; lea/m M.) y,  6740/ {777) Home;

pic and
Rerpr?r.

fills. II.
(at. 75.
to the
in
Parka-t.
50 from

nucleic
cam

'75.

tinand 

:liuenln?

lust, in 

3min

Vrnr. .

 

. 81-65 7"

SUPPRESSION AS RELATED TO TOXICOLOGY

- Author: J. G. Vos

If%??kt
- n3?
ouov

 

Laboratory of Pathology
National Institute of Public Health

Bilthoven, The Netherlands

Rafa-co: John A. Moore

Ratio-ll Institute of Environmentd Health Sciences
Research N. Park. North Carolina I

INTRODUCTION

Aberrant or defective immune responses are

known to be the result of primary immuno-
de?ciency diseases, in the DiGeorge 
(thymic hypopiasia) in man or in homozygous'

mice with the mutation ?nude? (rm/nu). which are
hairless and suffer from con tel th ic
hmlasia (Figure Immune deficiency can also
be secondary. eg, a result of undernutrition.
irradiation with or X-rays, neonatal thymee-
tomy, treatment with antiiymphocyte senun.
treatment with certain chemicals. and during
infectious diseases. Regarding chemical
compounds. relatively much infonnation is
available on the small number of immuno-
suppressive agents used clinically for immune
suppression. some thiopurines and slityiating

agents (reviewed by Bach).'
gurp?singlyu only a limited number of

 

chemicalntave bTen shown in toxicity studies _to
Hie?'munosuppresswe EmmesLang articu-
IaTiy the Ell-mediated immunity?haigeen Edy
smiled. A likely reason for this fact is that current

res for toxicity testing? underestimate-

importance of the immune system: 

 Organs and in general the immune system have
been poorly examined. 1heret?ore, in-depth

iagzzoun
- erogegomoa .

investigations were performed?with only a few
chemicals as to their effech the immune
response. This raises the question of whether these
chemicals represent only the tip of an iceberg. a
question which adequate
toxicity testing that includes careful examination
of the immune system both morphologitu and
functionally.

in this review, it will he stressed that knowiegde
from the expansive growing immunology discipline
thouid be incorporated into the field of toxic
eulogy. First. the immune system and Ire
immunological response will be introduced. Next,
attention will be given to the interdependence of
the immune system and the macrophage system.
Of special importance in toxicology is the
discrimination between direct effects on the
immune .system and indirect effects such as
nutritional de?ciencies, pathogenic organisms, and
changes in the endocrine balance. in this light,
procedures are presented to detect inrmuno-
suppression in routine toxicity studies. Further-
more. function studies will be discussed. of the
cell-mediated home system. the humeral
immunity, and the macrophag: system: these
function tests are meessary in order to gain insight
into the mode of action of the chemtal and to
determine the functional significance

?1
?11977 

a


u-u'ng' .


W. .. .

iymphoid organs. that increase the nrsceptibility
to infection. or that impair the immunological

responsiveness. These parameiers are considered

hnportant in the evaluation of the toxicity of
'uicsc chemicals. Chemicals for which in depth

lnfonnation is available (including data on an .

indirect or a direct action of the chemical) will be
disclosed first. In addition. chemicals whose
eiToets require further investigations will be
reviewed.. 

01:00)

The compound 2.3.7.8-tetraehlorodibenzo-p-
dioxin [Table 3) is a highly toxic impurity um
may be formed during the production of
Buuoi-loi et at.? were the
?rst to describe. in addition to hepatotoxlcity.
seven: thymus atrophy in rats exposed to TCDD





95

with rats. mice. and guinea pigs have con?rmed

and extended this In
omttast to the adult animal. at and mouse pups
that unexposed during the perinatal period by
maternal .treatment-do not show major liver
pathology; while. severe effects are seen in the

organs. particularly in the thymus."

Liver lesions-are mild in the adult guinea pig when

-ounpar'ed with the severe atrophy of thymus and .
peripheral organs.? 

At ahown by Moore. and Faith? (Figure 7).
rats spaced during the pre- andlor postnatal
period by maternal treatment with TCDD had
significantly lower body and thymus weights at
weaning 'age. At an age of 39 days. pups exposed
only postnataily gained more weight than animals
exposed during the gestation and nursing period.
After 145 days. the body and thymus weights of

the fonner poop did not differ signi?cantly from -

 


I
a

a

a
I

II


 

moi-b

 

mmom?maw 

 

 

. .
a

. (Figure 3). Since this i'u'st report. several studies

TABLE 3

. i?rox 7 

The Elf-ct of on Swollen?: violation and on Various Innuologleheanaateu 

?110999011100?

lavivooriarltro

 

speci- u'aatsncat rennin: meet. lat. - 
an invivo Thymus atrophy 53.68.9345. 91-? 
Moose - in Vito atrophy +0 52. 6.8. 94-96 
Gm?t- lam 
lat In who lama n-i?oetoprotein -
Wm! Irmthocyu count -
Ileana in rivo Patiohcral count . 100
Guinea pig in viro Peripheral count 
Mouse in vino Mahala! ntonocyta count - 96
Induction serum 0. ti. and 7 globulin I ,6
Susceptibility Saimateila ban infection #0 tot 
haccptibility.peodotabiea infection . - 101
lat . In viva Delayed-type hyperaensitivity 53. 5;
Guinea pig in who Delayed-type a
In in also Graft as. heat activity 51
loose in viva Graft vs. host activity . +14 32, ?8
lat . in viva Rejection of skin silo-grafts - 52
Jim . in rim Rejection of skin aiingrafts 4- 51
Gain at; in the Antibody teaponae to tetanus toxoid 40' .68
Rat - - In vivo Antibody reaponae to bovine gamma dobulin 53
transformation by I'llvir'o transformation by HM - +14 52
in vitro transfers-nation by HM - 52
lat Ia titre

blowouton transformation ?Denotes the 1nercaacd susceptibility to infection and the apprentice effect on [ransom dd?: 
moderate: strong: -. noe?'cct. -

Home: atm-hy also: occurred in adaenaioctoenbd and hypophysectotniaod rats can a: laica Heel-d with our-inc
'hunaone' 
162.1.8
?la young and adult anti-halt. instinctively. - 
'01: the [winery and smoothly antibody-response. manly.

 

 

It

 



 

 

wawwnuml

 

 

2

body walghl 

 

 

al

   

5

ill

 

 

J5.0.01
HS. um

m7. 
centroIsJet? female Fisheries: thatvereeapesod In


'the dose that caused atropliy of the thymus. In
contrast, TCDD had no gn scant effect on



monocytc counts. but serum eortcenttatims of a.
d. and giubulins were reduced by TCDD est-
posure.

A sensitive parameter expusme is

gum 
. lntubatlon of mice. once weekly rm} "gush gm;

as low as ATCDD per ram bod -

.wct' tl talit'ratc

and decreased the time from infection to death in

 


animals infected with Sell: or This dose
0 was one order of ma 'tu lots-er than

many in mice infected with pscudt?a?bics 
11m:
to infection is probabiy due to- the
endotoxln content of the bacteria, since recent
results have shown that oral intuhatlon, once
weekly for 4 weeks with 50 pg TCDD per kilogram
body weight (a dose that gave a moderate thymus
atrophy); rendered mice 50 times more sensitive to
endotoxln com."

01' the different parameters of the cellular

the delayedotype hypersensitivity to

postnatally via -7


l4 aodpre- and postnatally statuamaldosiuon
gestation 14. (Adapted front


tubercan was reduced in guinea pigs but not in
young adult rats.? The delayed-type hyper-

sensitivity to oxazolone was reduced when rats

the control. whereas scvere.;rowth stunting and

thymus atrophy were present in the latter group.
_Pra- and postnatal maternal treatment in mice can
also produce a wasting pups." These

?ndings clearly demonstrate the severe con- 
sequences of TCDD exposure during the

developrnental phase. 

The effect on the thymus la- not indirectly
mediated by- the pituitary or adrenals. since
thymus atrophy occun'ed ln'-'TCDD-exposed rats
that were adrenalectomized or_ hypophyseeto-
mined." In addition. the In?uence of TCDD an

intake.? I'Also. recent results have drown that
serum o-fetoprotein levels were not elevated rats
with TCDD-inducpd thymus atrophy." Finall
relirninary lentils indicate that injections of
"hormone" did not protect
TODD-exposed mouse pups from developing
thymus atrophy".1 did reduce the number
of peripheral in guinea pigs and mice.
but not in rats.'? In another study with mice."
no elfect was seen on peripheral and

I1 - CRCOirtrt-lkaiwhrm 6



?the thymus is not. caused by reduced food 

were exposed during the perinatal period by
maternal treatment with TCDD: this reduction was
even present in rats evaluated at 445 days of
age.? The pal! vs. host activity of spleen cells of
rats exposed .to TCDD during the postnatal
period?. and of young mice" was signi?cantly
reduced. but no reduced activity was seen when
spleen cells of adult mice were used." in both
rats and mice. perinatal exposure to TCDD
prolonged the rejection of skin transplants.?

Regarding the thymus-dependent antibody
response. treatment of guinea pigs with TCDD
reduced the secondary response to tetanus toxoid.
but no effect was seen on the primary response."
In the rat, perinatal exposure to TCDD did not
reduce the primary and secondary antibody
response to bovine gamma globulin."

The mitogertic response of 
thymus and spleen of rats enptned during the
perinatal period was suppressed on a cell-fur-ccll
basis and in particular on an organ-for-oman
basis?" The transformatim of spleen cells by

PHA was reduced in young mice. but not in adult

animal?s-a
The responsiveness of mane spleen cells and
rat thymus cells to the ultopens mm or Ciro A

.13. oa?

-- UtnanIn."

 

. .. huts. -u-r manna-Juana gar-11.



19

 

JJ

 

a .
 

I.

was not reduced when cultured in the presence of
TCDD at concentration: up to 0.02 
medium.? liven the presence? of 0.32 as TCDD
per milliliter medium has been reported not to be
toxic for human However. in
another study. TCDD at a concentration of 0.48

medium was cytotoxic for several mnm- -

'rnalian cell whereas concentrations that
are two orders of magnitude lower induce aryl
hydrocarbon hydroxylase activity in human
i'l'hese results make it unlikely
that TCDD needs metabolic activation. but
indicate that the depletion of the
thymus and the thymus-dependent lymriroiti
tissue is not caused by a direct cytotoxic action of
.
In conclusion. TCDD causes thymus atrophy in
all mammalian species studied. Regarding the
biological significance of this effect. it has been
shown in several experiments that TCDD exposure
raults in functional impairment of the cellular
humanity. but with quantitative different effects
on of cells. In particular.
T-heiper cells. possibly with the exception of those
in the prinea pig. seem less compromised. in
addition. it seems that the suppression of the
cell-mediated -immunity is an" age-related
phenomenon. at least in the mouse and rat where
- TCDD exponrre during ontogenesis of the immune
system seems to be prerequisite: in this respect.
TCDD mimics the effect of neonatal thymeetorny.
As discussed earlier." it is tempting to speculate
to what degree depressed cell-mediated immunity
in young rats and mice and in adult guinea pigs
contributed to their deaths. since severe effects
were observed in the absence of- major liver
apathology found in adult mice and rates-Regarding
I the mode of action of the TCDD-induced atrophy
ch the thymus. it is probably not caused by a
direct cytotoxic action on neither
!indirectly by. an effect on pituitary. adrenals. or
iproduction of thymic ?humane.? nor via in-
duction of e-fetoprotein or reduced food intake.
However. as discussed by van Lngten et al. it is
still possible that TCDD may impair the ab-
sorption and transportation of nutrients. An
alternative hypothesis that needs investigation may
be thruudt_an effect of TCDD on bone marrow
stem cells as precursors of in the
'lhymus. The increased susceptibility of mice to
infection with bent is remarkable. liost

resistance to this type of infection pri .-
depends - upon cell-mediated imnrunit 9

g. - .uL

however. since increased mortality occurred at
dhse levels of TCDD that do not produce 
atrophy. another mode of action seems likely. This
increased susceptibility is very probably due to the
endutoxin content of the bacteria. which has also
been demonstrated to be the cause of the greatly
enhanced susceptibility to E. infection of rats
exposed to Regarding this sensitizing
effect of TCDD for endotortin. it would be of
interest to Investigate whether TCDD impairs the

endotcxin detoxifying properties of macmphages. .-

Di-n-oetyltindichloride (Dom and Di-n-butyltin-
dichloride (one) -
ln-depth investigations were carried out on

effects on the immune system of the chemicals 

(DOTC) and di-n-butyltin-

dichloride (DBTC) (T able 4). As shown by Seinen 
and the main effect of DDT in rats is -

on the thymus. llistologically. deple-
tion was observed in. thymus and thymus-
dependent areas of spleen and nodes. in cell
suspensions of the thymus. a dose-related
decrease was seen in the number and viability of

- thymocytes; the effect an spleen cell count and'

cell viability was less pronounced. whereas no
effect was found on bone marrow count and

viability and' on peripheral and

monocyte numbers. .
As shown in Figure 8. in a series 
compounds tested, both' DOTC and DBTC pro-
duced a severe thymus atrophy (which was revers-
ible upon discontinuation of exposure). whereas a
less pronounced effect occurred with 

. chloride and di-nopropyltindichioride. in contrast.

the other compounds 
chloride. di-n-dodecyltindibromide.
octadecyitindibromide) as well as mono-n-
and
tetraoctyiti': did not cause- atrophy of the

- thymus.? - -

The selective effect of DOTC on thymus is not
indirectly caused by stress-related release of
glucocorticoids, since a similar effect on thymus
was present in adrenalectomized rats.? A strong
argument in favor of a direct action of DOT and
DBTC on rat thymus is their in vitro cytotoxicity

for of rats. but not of guinea pigs and

mice; in these latter two species. thymus atrophy
does not occur after exposure to these
chemicals.? . 

In function studies-of the thymus-dependent
immunity, the delayed-type hypersensitivity to

Hay 1917 OJ



.
v?d-MmH-H haunts-b.40th .MmJ-hJ-l-Iu- Ll-dloa-p- J..- -e nus ns-aeneu-an-I-Ieu .-

and di?n-_

 

Tun?a?p-n?po 
. 



 

 

. r-v?v 

 

"1

 

 



 

$8220

EXHIBIT 

KURON Complaints:

1. E. L. Hand - Lake Jackson, Texas
2. John Norton - Glendale, Arizona

azazzoN?

059583 Moo.

2

851

18221

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.s rm 7/
. .Quajr??w/ . 
Copies to: DSM 
Product Sales Manager
v: Product Tech ical Specialist
AGRICULTURAL DEPARTMENT Salesman
PRODUCT QUESTIONNAIRE (PLANTS) 
information given here should be factual unless indicated otherwise. 0
Written by: R. . Swart Date: 8/1l/71 
00
Exact Name, Address and Telephone Number of Complainant: . . Hand CD

Highway 332, Lake Jackson, Texas 77566 53L
Telephone No. 713'798'2590 County Brazor?ia State Texas
Detailed location ofdamaged and non-damaged area (locate on Geological Survey, highway, SCS or irrigation map):
Buffalo Camp Bayou north of Highway 332 - west of Lake Jackson,
Texas
Name, address and telephone number of person receiving complaint: - - MCGr'egor';
Agricultural Department, Dow Chemical Lake Jackson, Texas
Who requested Dow assistance: Mr . Hand
Name and address of Dow and other personnel involved in investigation:
D. M. Turner) Dow Texas Division Water Supply Deot.L Freeport
TR. w. Swarti Dow Agricultural Deot.. Lake Jackson
Date of complaint: July 1?2 1971
Product and Application: I
June El, 24, 25 Within
Product: Kuron Date of Application 1971 Date of purchase: 6 MO .
Proof of puchase: Distributor or Dealer: yo 0, il?e from DOW
Condition of container immediately prior to use: Lot number 50" 12 ?29
Amount of Product Used 55 Ga 1 - Area Treated- Rate/Acre Spray Vol. Spray/Acre
5 Gal. Plyac Emulsifier
Wind velocity and direction at time of application: None
Obtain original container with suspected contents: Spray water source:
Prior use of sprayer: NUDE Other pesticides used in tank mix: None

 

 

identi?cation, exact name. telephone number and address of:

Cmumam: Soravco Systems. 1327 Judiwava Box 10025. Houston: Texas 
Mr. M. w. Held - 715-686?0598. 18322

Page 1

Sub-contractor:

 

Contract applicator:

M00

Weed commissioner:




w.
4P

Farmer applicator:

Other:

 

Equipment used? method ofapplication (ground, air, boom or gun, nozzle, pressure, chisel. tarp. etc.): PPS 3 UPS
method of mixing (nurse tank or single batch): Spray from Boat

[5 equipment used for application of other pesticides?

 

Application Site:

 

 

 

Acres Crapland Acres
Utility ROW: _Crop:
Future
Highway ROW: Fallow: Crop
Railroad Row: I Grassland $1531.35 

lnd. Plant Site: 

Other: (Acres or sin of structure): Buffa Camo Bayo_u__? got 
of aquatic weeds 
Organism Affected:

Human?: Livestock:__Cattle Wildlife: Ornamental:

 

Crop: Speci?c identi?cation:

 

Number (plants, acres, animals, etc.): I 3 ions Dollar value Claimed: None

 

Proximity totreated area: . Cattle drink fI?QtTl bg yog_
'NoIif Dow Medical Director (Dr. H. L. Gordon): 2030 Dow enter, The Dow Chemical Carnpany, .liidland, Mich 48640- Phone (517)636-2105

Suspected Mechanism of Movement:

 

40,000 ga1./
Drift: Volatility: Water movement: __mimte_30il movement:

 

Crop residue: Manure: - Faulty Appln. Other: 
Field History (previous years):

StISpected pesticide: Date 
Other pesticides: Date 
Disease. insect or other pests: I 

Fertilizer (analysis): Source:

 

Source ofdrinking water, if sick or dead animals involved Bayou Whe I?e we re spra yed

Previous crops grown in ?eld

Soil type __Type ofirrigation employed 

Ht
CW
0%

co

Page2

Summary of Possible Contributing Factors Not Covered Above (including competence ofapph?cntor, other pesticides applied
in area. and opinion ofwrilcr as to cause): .

MrL,Hand is a retired Dow employeei_living on 5? acres adjacenttj
to Buffalo Camp Bayou. He has 16 cows and bull that pasture on
the area and drink from the bayou. The bayou carries water to the
canal that supplies water to the Dow plant. It is a practice of
the Water Sunply Department of Dow to spot spray weeds in this
bayou each summer.

.Mr. Hand stated he has had 4 or 5 calves die soon after birth
over a several year period, and had 3 abortions this year.

8129688 M0

1910 Bayou sprayed on June 29_and There was 1 calf died
soon after birth in the fall, several months after spraying.
1971 - Spraying was done on June 21, 2h, and 25. Calves lost:

 

 

1 . June 5 lived short while
1 June 27_ about 6 mo.
1  9 

 

 

I discussed the happenings with Mr. Hand and thanked him for

reoortinq to usL However, I do not think Kuron was the cause of

his problem and Mr. Hand really doesn't either. Various causes
of abortion were discussed, and it was suggested that he obnsult

his veterinarian with the possibility of checking blood samples.
He agreed to do this and thanked us for our interest, and will
report their findings to us later.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

l8224

Page3

SKETCH (to include the following in respect to area showing injury):

(I)

Page 4

Location oftreated area showing damaged and non-damaged areas.

Direction of surface water drainage.

Location and ?ow of nearby streams and/or drainage ditches(with distances).

Prevailing wind direction. 

Include location ot'other sensitive crops in the area and indicate whether or not affected.

Have photOgraphs taken, ifappropriate, showing overall effect and closeup ofalleged plant disorder.

 

{Vi/9683 MOO

16225

Summary of Disposition: of Claim or report:

Samples (samples to be taken only as a last resort). ll? independent laboratory, obtain:

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Name Address 93


Taken Analized Analytical results: 

Soil 


Water ES
+3

Plant CH

Animal

Wildlife 

Settled. closed: Nature of settlement: None

Complaint active: Recommended action: None

 

 

 

 

Mr. Hand, being a former Dow employee, said he thought we would be
interested in checking any possible relation of our product to his

problem. I assured him we were interested and again thanked him
for calling us.

18228

Page 5

u? -q

. munmoh?
- .

I
- 



.m New Scientist

. . 1 

. d?

13 April 1918

Chloracne: the chemical disease

 

Chloracne is caused only by contact with chlorinated organic chemicals?it is truly a disease
of modern civilisation. Although sometimes dis?guring. it is. In itself. not serious. But it could be a vit:
indication that sufferers have been exposed to chemicals with other, much more serious, effects

 

One cannot usefully discuss chlor-

Dr Kenneth Crow
is consultant acn'e
dermatologist at
Princess Margaret
Hospital. Swindon and
is a leading world
authority on chloracne

without considering the
chemistry, toxicology, and even
the environmental impact of the
chemicals which cause the disease.
Chloracne is a cutaneous eruption
which is generally 
but sometimes dis?guring. its sig-
nificance is as a marker, an indicator of the presence of a
chemical which is likely to be toxic in other?ways and may
well have environmental effects. When chloracne has
sounded its warning. our whole attention should be
directed to these other, more Serious, problems. They may
not be present-indeed generally are not?but until there
is conclusive proof to the contrary. chloracne should always
be regarded as a sign of potential systemic poisoning and
environmental hazard. Chloracne is produced by skin con-
tact, ingestion or inhalation of certain chlorinated aromatic
hydrocarbons. 
. thointact skin._ but in__doiue_sa_thcit._l_r_tx_ie.
eiiect is_les_sened._so_ 
tart 

Clinically. chloracne is an eruption of blackheads, usually
accompanied by small pale yellow which, in all but
the worst cases, are from pinhead to lentil size (above.lcl: 
in severe cases there may be inflammatory papulcs and
even pustules (a papule is a spot, 'a pustule is a pupulc
containing pus). The distribution of chloracne over the
body is interesting: the disease has a predilection for the
skin of the face, especially in a crescent outside and under

. . I. I . .Ithe eyes (the malar crescent) and behind the ears. Thor
areas are frequently allotted when every other part of U:
skin is normal. The remainder of the face, neck, shoulder
genitalia. chest and lower trunk can also be involved (i
that order). The hands, feet and legs are rarely involvr
and then only in the worst cases. in cases of exception.
severity unusual ?ndings may be' encountered. Ft
example. every single "hair" follicle in affected areas ma
become a blackhead or acne (above, right). in rarr
cases the entire body surface may be covered with 
horny spines like a nutmeg grater and in others generalisc
hyperpigmentation may occur'to such an extent that th
patient's racial origin may be in doubt.

There is every reason to believe that the response 
the human skin to the same dose of causative chemical ma
differ greatly in dill?erent areas of the body but this h.
never been tested experimentally. 1n mild cases. chloracu
may closely resemble adolescent acne but the curioi
concentration of blackheads and round the eyes an
on the cars is specific to chloracne. Its duration depeni
greatly on severity?the worst cases may still have actn
skin lesions 15 years after contact with the acneigen
chemical has ceased. Mild cases. however. clear with
months; after three years all but a "hard core" of 20 pl
cent of cases are likely to have resolved themselves. Tl
only permanent skin eifect is scarring. which may 
occur at all in mild cases but in the worst is very sever

Apart from the human subject, chloracne (or somethii
very like it) has been produced in only three experiment
?animals; the rhesus monkey ion the face, with loss of 

1822?

New Scientist

13 April 8  

cause the initial damage was due to chemical
the airborne dust consisting of particles of the
released by the accident. This dust produced 
and swelling seen in so many of the cases after
sion. As the burns faded. so the chloracne at
- almost every case it has been mild. Contact wit
these cases appears to have been mainly ext?.-
?rmed by the fact that (apart from the classic is
already described) only externally exposed arc
body were affected. All chloracneigens appe-
absorbed through the skin. although the extent 
tion varies greatly. Some systemic level of the
invariably occur. But such systemic concentratio

"c

 

 

 


a? 
-



 

 


1" .

7?

.a-o A

.0 ,?mmr



chlorines all result in the relative nonotoxmty 


. 
.cannula-ulna 
'3 1 I cloak- c}
. 

No available (top right). asymmetry (second from
top. right). masking of the reactive bridge by chlorine alums
(third [ram top, right), and. unadjacent chlorine atoms {botwm
right) all make the chemical relatively non-toxic

(causing steric hindrance) and the absence of adjacent

com-
pounds on the right of Figure 3.

Usually short- and medium-term toxic effects are only
seen in heavily poisoned subjects with severe chloracne
and, with few exceptions, have been produced by poly-
chlorinatod dibenzodioxins and clihonzo?
furans. The worst incident of poisoning by 
chemicals was the Yusho incident in Japan in 1068. when
over 1000 people were accidentally poisoned by consuming
a rice-based cooking oil contaminated with a poly-
chlorinated biphenyl (PCB) which contained an abnormally
high amount of dibenzofurans. it is now
believed that the dibenzofurans were largely responsible
for the toxic e?ects. The of severe poisoning are
known in detail only for TCDD and the mixture
in the Yusho accident. The two chemicals differ little in
their target organs. Chloracne. disturbances of the central
and peripheral nervous systems. wasting, hepatic damage
and hyperlipaemia (high blood lipid levels) have been most
prominent. Recent experimental work revealing that
TCDD. although known to cause birth defects. is in fact
far more directly toxic to the fetus. is borne out by the
history of pregnant Yusho victims: thWit-e
incidence of st'l - ir 
dioxin-liktLL?CDFs.
Wknown lob; fetotoxic but not teratogenic.

An interesting toxic manifestation is meta?
bolic abnormality in which the pigment is re-
tained in the tissues. For years his been known
to be associated with severe TCDD poisoning in man and
can be produced in experimental animals by TCDD and
PCBs?-?even pure PCB isomers. especiallv the
hexachlorobiphenyl. It now seems most likely
that may be due to an et?lect on the enzyme
delta-aminolaevulinic acid the enzyme that
limits the rate of production. This is
strongly induced in vitro by TCDD and it seems likely that
the PCB isomers which strongly induce AHH alsi) induce
the enzyme.

Mild chloracne. such as that suffered after the much-
publicised Seveso accident in Italy in July 1976. does not
appear to be accompanied by short-term systemic 
and surveys of the Seveso population one year after the
accident con?rm this hypothesis. The absence of terato?
genie effects at Seveso is as expected in view of the ?nding
already quoted. The Seveso cases were most confusing be~

are unlikely to cause problems. except possibly i:
term. when much depends on rate of metabolism.
here is no possibility of consumer poisoning r,
mild chloracne from maliu??_c_t_ur_ec_l products. 
the chemical?"industry the risk of chloracne
although much diminished as awareness of the
increases and-'? as diagnosis (especially in mild 
comes more accurate. From the environmental 5!
the risk of low dose. long-term effects?in particul.
and damage to genetic material 
ably more important. Poorly metabolised materials
a hazard in the long term even when the dose is sm.
Experimental data on the long-term effects 
acneigens on animals are limited to the rat and
When fed to rodents in large doses throughout ll:
time. PCBs produce liver tumours. The results of .
study in which varying amounts of TCDD were fed

_throughout their life revealed that tumours were

only by a dose large enough to produce other cvid

chronic toxicity in various organ systems. Lower

which did not appear to affect the health of the 2

produced no tumours. This suggests a dose-relates

Of further interest is a careful follow-up of 70 mt
severely poisoned by TCDD 24 years ago in an int
accident. So far there has been no evidence of ex
or unusual malignant disease. in it seems 
may be carcinogenic lo maiT?c
high doses. There remains ot addlt
synergistic effects between mild carcinogens. as 
co?carcinogenesis. when a non-carcinogen can be act
by interaction with other chemicals. As yet noth
known about these phenomena among chloracneiger

these are under intensive scrutiny following the race:

covery of in a wide variety of materials.

Controlling the risks
What can the chemical industry do to reduce the
of chloracneigenic chemicals? One priority must be t:
lish and disseminate toxicological information i

.throughout the industry. Otherwise costly and 1e

research would be wasted. Secondly. because acne
potential in a chlorinated or brominated armuatit
stance appears to imply various other toxicities whicl
be serious and of environmental signi?cance. 1 believ
the simple. but extremely sensitive. rabbit ear test 5
become part of the standard battery of toxicologicd
applied to every new substance (especially lierhicidr
pesticides). If a contaminant is responsible for the a
it may be possible to eliminate it altogether or reduc
negligible levels (providing that long-term low dose

have been adequately assessed). The headlong 
produce new chemicals after the Second World War

that a great many new substances were inadeq
examined toxicologically. Increasing awareness on ll
of the major manufacturers and government researcl
tutions. especially in the US. makes such problems it
less likely to recur.

This whale to band on a super delivered in: weak to m. on a o!
Cocoon-amt Huron. Society. .

 

 

 :l  New Scientist 13 April 1978  iavy}
disruption of glands in the eyelids?Meibomian glands  

 'w?u In the human subject); the hairless mouse. and, most
.  of all. on the inner surface of the rabbit?s ear.
I Berg?s-ll ll not possible to produce ehloracne on any other part
I ?g (453: the rabbit's skin. Indeed, even systematic absorption of
i1? {7 chloracneigrns by the rabbit produces chloracne only on
 the inner surfaces of the ears. Thus there is a clear parallel
5?12 :Llo the curious sensitivity of the molar crescents and ears in
the human subject. The rabbit's ear appears to be the most

- it? emittive biological surface known, reacting to a total dose

in I  the most potent chloracneigenic chemicals as small as
.. microgramme. Thus the rabbit's ear affords a simple,
an and relatively cheap method of identifying chlor-
mCiScPiC materials. The_biolegical basis of chloracne, and
 fair. particular of its predilection for certain areas of the
Lil

  

is quite unknown. Investigation of the disease is
*ii'thmpered by the lack of an analytical method accurate
band sensitive enough to estimate quantitatively chlor-
- 

r31, acneigenic chemicals which may be present in blackheads
1i}, glad as opposed to other skin tissues.

The chl::inated aromatic hydrocarbons which are known
-1559 to have produced chloracne in humans are

?He

13;, 
.3 511.3," commercial (PCBs)

dibenzofurans 

dibenzodioxins 

I tetrachloroazobenzene (TCAB) and tetrachloroazoxyben-

'35! zene (TCAOB).

With the exception of the chlornaphthalenes and PCBs.
- '21" these Compounds are contaminants formed accidentally
.43.. .. 'during the manufacture of other chemicals. For example.
the PC85 have been shown to have a toxicity of
1? their own, in their commercial form they are always con-
:t-?taminated by varying amounts of the much more toxic
Again. the manufacture of 2,4,5 trichlorophenol
"'g?produces as a contaminant the most toxic and chlor-
.: .jacneigenic small molecule known, 2.3.7.8 tetrachlorodi-
"JbeHZD-p-dioxin (TCDD) of Seveso fame. Crude pentachloro?
(if it phenol production. as recent research has revealed. leads
- the formation of a range of impurities. The latter
also recently been discovered in crude hexaclilor-
I f3? -benzene. The contaminants TCAB and TCAOB occur during
-: . "$51 the chemical reduction of to the cor-
responding diphenyl hydroxylamine and dichloroaniline,
.both industrially signi?cant intermediates. TQM) and

9-3
sf"



near-





4 

sari.



  

 TCAOB are never found in the final products and exposure
?If, therefore occurs only in the chemical industry. Little is
  known of the detailed chemistry of the commercial chlor-
q' naphthalenes. Whether their toxicity is due to contamin-
. ants or the pure chlornaphthalenes is unknown.
 The acneigenic potential of a chemical appears to he.
 fr; ,r directly related to its overall toxicity. The importance of
.3 3.1 this fact is the implication that chloracne an expert-
h:  mental animal can provide a very important screening
.f g- test for systemic (not only skin) toxicity. This appears to
1 hold true for all known chloracneigens with the exception
of tetradiloroazobenzene and possibly azoxybi-n-
- zone. Recent work reveals a vast difference between the
 oral and cutaneous toxicity of these chemicals. presumably
because of the well-known susceptibility of the azo bond
v- 13}, to destruction in the mammalian gut.
- Certain basic elements of their chemical structure deter-

 mine 'the toxicity of chloracneigens. The crudest g_f__all
factors i?_t_l3e_degree of chlorination. It appears that a

 

1' 

2.1.1.: 

[y 110 
1m Uri.?

(1.71?How carbon atoms i
in aromatic at
hydrocarbons or

numbered i'
 
chipping gentent somewhere between ma um and mi

mum produces the greatest toxicity. For example, 
dioxins can have from one to eight chlorine atoms. 
than four, or more than six. chlorine atoms in. a diox

molecule greany reduTc t'oxiCiE betwden tour and?six pr
Wm??p thew
eiemicat property determining toxicity is isomerism, th

is. not 'ust the number alogcn gt_o_ms_b.u.t__thcir_m

posnion in th?olecule. By convention these positions a
allotted a number (see Figure 1). Thus we may ha-
2.3.7.8 and 12.3.4 tetrachlorodibenzo-p?dioxins or, similar:
and tetrachloroazobenzene. 


must be able to distinguish them. Ilihe detailed study i
as many isomers as possible is essential. Successful we:
in this ?eld depends entirely on the Chemist's ability i
the various isomers in pure form. Once the-
are available as standards, modern instrumentation pr
vides a means of relatively rapid. accurate and sensitix
analysis, not only qualitative but also quantitative.

 

Simple test?tube test:

ironically, many toxic chemicals initiate their (in
destruction?they can activate (or ?induce?) cellul.
enzymes whose function is to metabolise and detoin
them. These enzymes can be separated from liver cell
a particularly rich source. and used for in vitro tests in
tone potential. hydroxylase 
the most useful for assessing acneigenic chemicals. Th
degree and duration of enzyme induction has proved 
parallel closely the acneigenic and systemic toxicity of a
rhloracneigens so far studied. giving us an elegant an
simple method for distinguishing toxic from less tox'
isomers. On the other hand. in vitro tests will not reve.
the sometimes great differences in susceptibility betwei-
species. Thus animal studies are still necessary but onl
for a very small number of selected compounds. As a rcSul
of recent intensive research with the enzyme inductin

'"i?m?luc 


in the molecule. the 
adjacent carbon ate 5. the presence of available hydrogen
the Spain-
;irraneenient of a molecule "hides" reactive atoms. pn
venting certain reactions) all enhance toxicitV. tlecen
work indicates that all ?ogcireiggiic- compounds _ai\
either exactly or approximately isostcric (that is. have th-
same distributions of electrons?over the outside of th-

melee, le?see Figure 2). The 
erat -?asimmetry, chlorination next to the diphenyl bridg-

 

 

l.4 . 

tease

1,3,1. I THMCEWMEZOFWN





 

. - 

 

 

- I  
Pest-lame: necuaarnon owmon euaacr IURIAU no. era?Rn?
enumerate. o.c. zozuo 
FOR AMENDED REGISTRATION OF ECONOMIC POISONS LOAN: or APPLICATION 

- .. . A ..
(Under the Federal Insecticide, Fungicrde, and Act) ?a ?l 0? -



 

 

IMPORTANTI READ msrnucrraus on REVERIE

2. "mm" (Must be same product name as on label?do not list active ingredients)" I :311.3.5.4. 

HURON

 

 

 

 

 

3. [runs a MAILING ADDHEII an neatsrruu'r (Include Zip Cock). I . 1 ?Ineatsrni?ri?arr no. 
-2 Dow Chemical - :4 
Ag-Organics Department "l l. ?I?ll- DATE 03
NC
Page   3  . 3:5: r1   
Michigan -  . 33.373  r12. 'Lu 2.5: 
lAItn: Dr; V. I?ullison "Treats-distal}! (V
6. NATURE OF REVISION (Cheek applicable item and give details in item 7, when required)
"Elmo" OF LABELING q] . usle's'l'raoea {o LAalLlhia i 
FORMULAHON [3 {List new recommendations in item 7 (5?55
(Give description of exact change ,h . .   ape-Hence encoucr NAME Lb 
in item 7) I  I .U   .i   ram.? 91?; name and new name . 5% 
OTHER a . .. L..-. 1. V.  in item 7) -r  I    
 in item  .7 'l 7?lif 1 . . hrs-2r:- a-J- 
7. oc?rmu acauunco av nzvmou cuecxeo new 0 (Attach additionaisheets if more space is needed) .  

This application for amended registration involves a general label re?
vision. The recommendations have been reorganized into four major 
headings: Industrial and Non?Cropland Areas, Aquatic Weed Controlg'?
Pasture and Rangeland, and_Uses in Croplands. _The_precautionary state-
ments have been expanded and strengthenedreally new uses have been added but the label does have several recom-
mendations which are practices that have been in common use but were not
previously stated on the label. "These are} forest conifer release,
basal bark and stump, powered knapsack_basal treatment; and modified
basal treatment. These recommendations are-all for Industrial and.Non-

Cropland areas. i' . -- .u.v . .. . n.
EELThe Aquatic section has been modified td'make it shorter and
less complex. 

commune on ATTACHMENT 

8. THE FOLLOWING MUST BE SUBMITTED WITH THIS APPLICATION IN ANY CORRESPONDENCE ON THIS PRODUCT REFER TO-

REGI THATION 1H ITEM ABOVE
0 Five (5) copies of revised labeling, including any printed or graphic matter 5_ J, . l, .-  
- 

which may accompany the sale of this product. Copies must be clearly -  
legible and identical. ..
If a change in formulation is involved, five (5) copies of a stotemenr or
revised formula showing the precise name and preaentage of "5h active
and  inert ingredient.
(This information is treated confidentially)
When appropriate, three (3) copies of Supporting Data.

9, SIGNATURE OF AUTHORIZED FIRM REPRESENTATIVE.-


m. TITLE II. DATE SIGNED I,

 

 

 

ietrntien Qggcialist 
PR FORM 9-198 ISTIMG STOCK OF on (saws?n93 nus. 19w - .
My 1959 BE USED UNTIL exnausreo APPLICANT 5 COPY

?16230

. 3 

r1INSTRUCTIONS FOR PR man?s?its

--. cud?- 


- 'I?qI-rll-T-?I-?u4l"
"31w"- 13:? . F- 1231'!
2 2' ti .bApplications should be submitted as far in advance as possible prior to desired registration date. 'Thae'time
required to process applications may vary depending on the extent of review required. Applications which
 require consultation With other governmental agencies will take__a longer time _to.process.  

.
a - no n-ioi-rnuit-ruu cut. 5.1a 

2. ?'The name of the economic poison shown in the application must'be' the'same'pr?du'ct'nahie ",afs'that'shownm?
thelabellns submitted. Do not list the active ingredients. i  

i-

3. -?The name of the firm or person and address shown is the: person or; firm 
istration will be issued. If you are acting in behalf of another party, yen must' submit'authoriiation?from 
that party to act for them in'registration matters. The address given in item three will be the mailing ad-

__._dress permanently on record unless changed by the registrant. 
314-7 a tailifhcg?gngg'The registration number assigned must appear on the label. The number must be the'same as that appear-
ing on the notice of registration and shall be preceded by the phrase Registration NumberReg. No.9 (Better to Sec. 362.607) of the Regulations); . ?r - a ..

MOO



883683



i=3" it 331 nails .1  :3 2-13,; can melt n't?nu'?qqc .b
5' Changes in the formulamust be submitted in advance of-the prOposed effective
data rc-Jf-i it} 

?enthusiasm: in .3 DHAHNature of revision - The Whe exact changes desired and ?p?dn?reques?t must subf
mit a description of test results to justify such changes. Minor changes in formulation may be handleda's
revisions of existing registrations. This might include a change in the percentage of active ingredients or
 .-a..change, in inert.lngredients..However, any basic change in formulation such asachangein the principal ..
active ingredients would require a separate registration: as a?new product wi-th-a' different? product' nam'..-.--. 


 

.u .-


. . I t. ..-.. .. 
. i. 

-- . 1' 
B. Labeling All' cepies or labeling must be clearlylegible' and lithograph labels
should be submitted. Do not send containers. Your supplier should be 'able'to furnish photo copies of the.
screen print. Submit reduced photo capies of labeling for dry agricultural fertilizer '4 insecticide, or her-
bicide bags. Any label smaller than half the size of a sheet of paper 8 inches by 10 inches Should be stapled

to a sheet of paper 8 inches by 10 inches before attaching to application.

.- l?
-. .I

Formulation ?r If a change in formulation is involved, five copies of a saparate statement listing the cor:
'rect name and percentage by weight of each active and each inert ingredient must be submitted with each
application. This statement is treated confidentially. it cannot be used to support a customer's (formulat-
or) application for subsequent registration without written authorization from the registrant. If necessary
- information on the formula is not available to the applicant (formulator or packer), he should: Obtain
the necessary information from the basic supplier and submit five cepies of the statement with the appli-
cation or Request the basic supplier to furnish the necessary information to the Division along with
a statement specifically authorizing the use of such information to support separate registration for the
applicant. The statement from the basic supplier must clearly identify the applicant's product to which it
appliesATTENTION: The submission of an applicstion does not constitute registration. Comment
or notice of registration will be sent after examination by the Pesticides Regulation Divi-, 
sion of the information submitted?Euro-r, :irL. lo sit-.in: enu- AM 1;   i at  tun-'0
.u 1 - 'ng?i Mel-i 7:315:i?f?-.-.. H-. .. 
5 ?4 ?Six ?In. :9 '3 rel-w 
. .. a] a . ?ifWILLARD AND BILLEE SHOECRAFT

VS.

THE DOW CHEMICAL COMPANY

1 EXHIBIT 

2,4,5-Trichlorophenol Analytical and Shipment Record.

16232

 ANALYTICAL AND SHIPMENT RECORD MICHIGAN DIVISION, DOW CHEMICAL U.S.A. FOR 

ALL LOTS PRODUCED DURING THE PERIOD MAY 27, 1967 AND MAY 2, 1968 THAT WERE USED IN THE MANUFACTURE

 

-I48.11") 
:4 a: o? r: 
LOT DATE BLDG CAR (PPM)
05277 5/27/67 489 309 0.2 . . 91.5 0.2 .2 0.3 <1
06017 6/1/67 489 313 0.2. . .7 89.7 _0.2 .5 0.2 <1
06067 6/6/67 267. 314 <0.5 0.2 . .5 89.7 <0.5 .2 <0.5
06077 6/7/67 489 313 <0.5 0.3 . .8 89.2 <0.5 .4 <0.5 <1
06127 6/12/67 489 314 <0.5 <0.5 . .0 88.1 0.2 .0 0.4 <1
06137 6/13/67 489 313
   309.; 0.4 I3 0 .9 
06167 6/16/67 489 314 0.4 . . 89.2 .3 .1 <1
6/23/90.8 . . <1
0;
03

OQTSTOMOO

I

SHIPMENTS OF TRICHLOROPHENOL

 



-.-LOT DATE. BLDG CAR (ppm)
I

06267 6/26/67 489 309 0.4 .5 91.0 . . <1 .
06287 6/28/67 489 313. 0.5 .6 . 90.5 . . <1
06287-2 6/28/67 489 314 0.4 .6 90.0 (1
06307 6/30/67 489 - 309 <0.5 <0.3 .4 . 92.7 <0.2 . . <1
07067 7/6/67 489 313 (1
07087 7/8/67 489 314 <1
07117 7/11/67 267 309 0.4 0.1 ,0.4 I 3.1 90.3 2.3 0.1 <1
07137-1 7/13/67 489 . 313 0.4 0.1 0.4 2.8 89.9 0.2 2.2 0.1 <1
1M07267-2 7/26/67 267 309 <0.5 0.3 0.6 2.2 92.6 <0.2 1.8 0.4
0508107-2 8/10/67 489 309C '0.3 0.2 0.3 i.6 88.5 0.2 2.1 0.3 <1
2308157 8/15/67 267 314 <0.5 <0.1 <0.5 1.7 87.9 <0.2 1.8 <1.0 <1
?h08157?l 8/15/67 489 313 <0.5 .1 .5 1.7 89.2 0.2 2.1 <1.0 <1

MOO

SHIPMENTS OF TRIUHLOROPHENOL

 

.
m'r-I
I I I - -.-LOT DATE BLDG CAR (PPM)
08167-2 8/16/67 489 309 <0.5 . 90.4 0. . <1.0 <1
08247 8/24/67 489 313 <0.5 . . 92.3 0. <1 <1
08217 8/21/67 489 314 <0.5 91.5 0. <1 Inter-
ference
08287 8/28/67 489 314 <0.s . 92.8 0. 0.3 <1
08297 8/29/67 489 309 <0.5 89.0 0. 0.3 <1
09087 9/8/67 267 309 0.4 . . 89.0 - . <0.3 <1
09057 9/5/89.6 0. <0.3 <1
09017 9/1/67_ 489 313 0.3 89.0 0. <0.3 <1
09117?1Da 9/11/67 267 294 93.5 0. . <1
09147 9/14/67 489 -313 - . 93.2 0. <1
09167 9/16/67 489 314 91.0 0. <0.5 <1
09207?1 9/20/67 489 313 - . 90.4 0. . <0.3 <1
a .
891810 M00
(29

SHIPMENTS QF TRICHLOROPHENOL

 

8839


-.-4 W4 'rN?s.
LOT DATE BLDG CAR (PPM)
09207-2* 9/20/67 267 '294 91.7 <0.3 <1
09227-1 9/22/67 267 294 <0.1 93.1 0.3 <1
09237 9/23/67 489 314 <0.1 92.5 0.3 <1
09267?1 9/26/67 489 313-0 . 92.2 2.3 0.3 <1
09287-1 9/28/67 489 314?c <0.1 93.2 0.2 <1
'09297-1 9/29/67 489 313-c <0.1 . . 92.2 0.4 2.3 0.2 <1
10037-1 10/3/67 489 314-0 <0.1 . 90.0 <0.3 <1
10057?2 10/5/67 267 294-0 <0.1 . 91.0 2.7 <0.3 <1
10067-1 10/6/67 267 294-0. <0.1 0. . 89.9 <0.3 <1
10067-2 10/6/67 489 309-c' <0.1 .5 90.0 0.2 2.3 <0.3 <1
10067?3 10/6/67 267 294-0 <0.1 .4 90.1 2.0 <0.3 <1
p.145.

691810 

 OF TRICHLOROPHENOL

 



4JE-4
0 0 OJ 0 0 01m'u

LOT DATE BLDG CAR a (PPM)
10097-1 10/9/67 489 313-C (0.1 0.5 0.5 2.7 89.6 0.2 A 2.0 (0.3 (1
10097-2 10/9/67 267 294-0 (0.1 0.5 0.3 2.9 92.1 0.2 1.9 <0.3 - <1
10127~1 10/12/67 267 309-C (0.1 0.6 0.3 2.4 90.0 0.1 1.8 (0.3 (1
10137-1 10/13/67 489 314-C (0.1 0.7 0.4 2.5 91.2 0.2 I 2.2 0.3 (1
1016741 10/16/67 489 313-C (0.1 0.7 0.3 2.9 90.5 0.2 2.4 0.3 (1
10187-1 10/18/67 489 314-C (0.1 0.6 0.3 2.7 90.5 0.2 _2.1 0.3 (1
10217-1 10/21/67 489 313-C (0.1 0.4 0.2 2.3 91.8 0.2 2.4 (0.3
'10237-1 10/23/67 7 489 - 309-C (0.1 0.3 0.2 2.7 90.8 0.2 2.3 (0.3 (1
10257-1 10/25/67 267 314-dv (0.1 0.4 0.4 3.4 89.7 0.3 2.3 (0.3 (1
10287-1 10/28/67 489 313-C (0.1 0.6 0.4 3.6 88.9
38817-1D 10/31/67 489 309-C (0.1 0.6 0.3 4.0 88.3 0.3 2.4 (0.3 (1
6%
ES
99
4191810 

SHIPMENTS OF TRICHLOROPHENOL

 



LOT BLDG CAR (PPM)
11027?20 11/2/67 489 <0.1 0.6 0.3 4.3 89.5 0.4 . <1
11067-1D 11/6/67 489 309-C (0.1 0.1 0.5 4.4 89.7 - 0.3 - <1
11097-1F 11/9/67 267 313-C (0.1 0.5 4.6 91.8 0.3 .3 0.3 <1
11137-1F 11/13/67 489 3.3 91.0 0.2 .1 (0.5 (1
11/24/67 489 309-C 2.6 90.7 0.2 0.2 (1
11257-1D 11/25/67 267 313-C 2.4 89.8 0.2 0.3 (1
11277?10 11/27/67 489 2.4 90.2 0.2 0.2 (1
12047?10 12/4/67 489 2.7 90.5 0.3 0.3
12087-10 12/8/67 489 309-C 2.4 90.7 0.3 0.3 <1
12097-10 12/9/67 489 2.5 90.5 0.2 0.3 (1
12107-1D 12/10/67 489 2.4 '91SQTBIOMOCI

m.

SHIPMENTS OF TRICHLOROPHENOL

 



'r-I 'v011'}
LOT DATE BLDG CAR (ppm) I
I
12147?10 12/14/67 489 314?c <0.5 0.6 2.4 89.8 0.3 2.3 0.3 <1
01128-1D 1/12/68 489 313-0 - 0.6 0.7 2.7 90.2 0.2 2.4 0.3 I <1
01138-lD 1/13/68 489 309-0 0.7 2.9 90.2 0.2 2.3 0.3 <1
01158-1D 1/15/68 489 314-0 - - 3.0 89.3 0.2 2.3 0.2 <1
01188-1D 1/18/68 489 313-c <0.5 0.8 3.3 90.5 0.2 2.3 0.3 <1
01218-1D 1/21/68 ?267 314-0 <0.5 0.7 0.7 2.8 91.2 0.2 2.2 0.3 <1
01278-1D 1/27/68 489 309-0 0.7 0.6 2.8 89.3 0.2 2.3 0.3 <1
01298-1D 1/29/68 I 489. 313-0 0.7 0.7 2.8 90.7 0.2 2.3 0.4 <1
02038-1D 2/3/68 489 .314-0 0.7 0.7 2.6 91.8 0.2 2.1 0.4 <1
02078-1 2/7/68 489 309?0 (0.1 0.6 2.8 91.7 0.2 2.2 <0.1 <1

QSISIOMOCI

89391

SHIPMENTS OF TRICHLOROPHENOL

 

W391

1.510
.LIEr-I r-I .-.-I -r-.-LOT DATE BLDG CAR . (PPM)
02108?1 2/10/58 489 314 <0.1 . 0.5 92.0 . . <1
02118-1 2/11/58 257 313 <0.1 . 0.7 91.0 . . . <1
02158-1 2/15/58 489 314' . 0.5 . 92.2 . . . <1
02208-1 2/20/58 489 <0.1 0.5 0.5 91.9 . 0.3 <1
02288?1 2/28/58 489 314 0.5 0. . 92.5 . . 0.3 <1
  03098-lF 3/9/58 257 309 0.5 0. 91.5 . 0.3 <1
03208-1F 3/20/58 489 313 0.5 . 92.9 . . 0.3 <1
03228-1F 3/22/58 489 314 0.5 . 92.4 . 0.4 <1
03258-1 3/25/58 489 '309 0.5 . . 92.2- . . 0.3 <1
03288-1F 3/28/58 489 314 <1
03308-1 3/30/58 489 313 0.7 0.9 2.4 92.0 .2 2.3 0.3 <1

LQISIOMOO

SHIPMENTS .1: .120
41J1:
3  \0 con11') 
6? m? 63?. 6: 2? 
LOT DATE BLDG CAR %v (PPM)
04028-1 4/2/68 489 314 0.7 . 2.4 89.9 . .4 . <1
04058-1F 4/5/68 489 309 0.6 . 2.2 91.7 . .3 . <1
04098-1F 4/9/68 489 . 314 0.7, . 2.3 91.5 . <1
04118-1 4/11/68 267 313 0.7 2.3 92.7 . . <1
04178-1F 4/17/68 489 0.7 2.5 93.6 . <1
O418851F 4/18/68 267 314 0.7 . 2.5 91.7 . . <1
04228-1 4/22/68 489 313 0.5 2.1 93.2 . . <1
4/26/68 2677 309 0.6 2.2 93.3 . . <1
04298-10 4/29/68 489 314 1 0.6 2.2 90.6 . <1
GATX
05018-1D 5/1/68 489 70116 0.6 . 2.2 92.1 . . <1
05028-1D 5/2/68 267 313 0.6 0.6 1.9 93.8 0.1 2.1 0.4 <1

cm
 891810 MOO



 ?yr12-.

h.
I



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I.
(X ?lL-t

030/352"  
030984ng 267 3307 

 

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03/38 44/ /2/2

 

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3

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33/5845? 265 P/pe- 0.6



A A h.

 

came-mg: 265 ?fe a, 6



 

4/93 3/3 a? 0.6



 

33223?? 43-7 3/4 I ?46

AAN

 

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was-J 433 332 0.6

 





3253 03258-3 4- 10

 







.43 x? 04.


 



 

 

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03368~l 33489 3/3 

 

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Vol. 13. 6. Xowmber. ??70

29

cry/Tm M00

18249

Effects of Herbicides on ??ater and Its Inhabitants?

W. R. MULusoxi?

Abstract. There is little evidence that herbicides from agro-
nomic or industrial usage are reaching or accumulating in
our water supplies in amounts to cattse a pollution problem.
Fish tolerance to weed killers varies with their size and species
as well as with differences in the aquatic site. There may be
variation to different forms of the aCtive ingredient. Other
Components in a formulation may be more toxic than the
herbicide itself. Therefore. the LD-st of different fonuulations
should be determined. In addition. the manner and Other
circumstances involting application of a herbicide may change
its toxicity to water inhabitants. Our current knowledge of
the effeas of herbicides on fish. plankton. and other water
inhabitants indicate that harmful effects with our present
herbicides, when such min. are only temporary. Available
evidence suggeSts there is no biological magni?cation problem
with herbicides.

Ixrttoot'crtox

ooxv there is much professional and lay intereSt

in our environment. particularly with to
pollution problems. The unfavorable subtle effects of
dichloro diphenyl trichloroethane (DDT) on certain
organisms and its praaically universal distribution in
our environment have stnuulatetl questions concerning
the possibility of similar effects from other agricultural
chemicals.

Herbicides are common today and their use is growing
rapidly. Increasingr concern will be directed toward the
effects of weedkillers on non-target organisms as well as
the ultimate fate of these chemicals in our environment.
Our analytical methods are becoming increasingly sensi-
tive and trace amounts of various herbicides are likely to
be found in more and tnore places. However, interpreting
the importance of such small quantities as parts per
and even parts per billion of the weed killers
when found in our environment is the real problem. It
appears timely, therefore. to survey the possible occur-
rence of herbicides in water and to summarize present
knowledge concerning their elfects on fish. plankton. and
other water dwellers. 

The residual effect of herbicides in the ecosystem that
involves water supplies is of great importance. Drainage
from watersheds and other areas treated with herbicides
might result in secondary effects involving the health of
domestic animals. wildlife. aquatic animals, and even
man. It has been estimated (90) that more than 
of the population will depend upon surface water for
drinking by. 1980. Thus. the prevention of water con-
tamination'is of the utmost direct importance to man.
\?r'ith the increasing use of herbicides on non-cropland
as well as cropland. the significance of their occurrence
med for publication October ill. 15:39.

Department, The Dow Chemical Company, Mid-
land, Michigan.

738



in surface water be evaluated. The accumulatioz
of herbicides. if it occurs, by plankton and water animal
could start the biological magni?cation process ?rst s.
clearly demonstrated with the insecticide DDT (63
Thus, their effect on fish, plankton. and water cruStacean
is of considerable consequence.

There are several general references dealing with [its
toxicity of chemicals. particularly herbicides, to fish
Applegate et ul. (5) and Lawrence (73) both have exten
sive fish toxicity data. Three reports from the Fish ant.
Wildlife Service (135. 136, 137) are excellent summaries
of lnvesdgations carried out by the US. Department ot
the Interior. The Herbicide Handbook (62) has fisl:
toxicity data for many weedkillers.

There have been relatively few investigations on the
effect of herbicides on plankton. Usually the work re-
ported occurs as a brief note in conjunction with a
herbicidal study of an aquatic weed problem. As the
Study of herbicides is becoming a more mature science
and we have realiyed the possible existence of a biological
magni?cation problem through herbicidal the, more
emphasis and study will be placed on this aspect of the
subject in developing new products.

This is not a review of herbicidal effectiveness on
aquatic weed problems. Klinmuan (GS). Fryer and livans
(-13), antl more recently the National Research Council
(85) in their books have good sections on the application
of various herbicides for aquatic weed control. A general
review on this subject also is given by House or til. 
and a good history of the development of aquatic weed
control is given by Timmons (139.). Instead. this publica-
tion is concerned with the ultimate fate of herbicides
after use and also their etl'ects on non-target organisms.
In this paper, the organization has been according to
the common names of herbicides taken up in alpha-
betical order. In certain instances. for convenience. ntan
than one may be discussed under one heading.

ERBICIDES

Acrolcin. Burdick at (l7) reported that acrolein is
very toxic to ?sh. The chemical has a S-l-hr of h;
for fingerling brown trout L.) and 71?
for bluegills (chomfs? macroefu'rus RA.

Ferguson ct (33) used acrolein to control aquatic
weeds and snails {Bu/inns and spp.) in the
Sudan. studies at 25 and 50- showed
minor or no injury to furrow-irrigated crops. but flood
irrigation of vegetable seedlian at 15 was toxic.
Effective carriage of acrolein was demon-
strated for a distance of 1 mile. The snails were US to
killed and all aquatic weetb were
dest-roved.

L'ur'au et al'. (133). experimenting with acrolein in

JMedian tolerated limit.

Volume l8, Issue 5 

18250

MULLISOX

also found it very effective against snails as well
5 the aquatic curlyleaf pondweed ti?onmtogetmt crispus'
_t in irrigation ditches. The 20 to DU concentration
.eciuired for weed control had no adverse effect on local
crops. Field te5ts demonstrated the importance of water
control during tlte application period.

Arsenic. .\lackentltun (77) reported on tlte long-term use
of sodium arsenite. During the past [0 years, lb
of have been applied along a lake shoreline:
uniform distribution of this quantity over the top inch
of bottom mud or hydrosoil would amount to 330 ppm.
.\lttd samples analyzed itt tlte fall had 48 to l5-l 
.and the following spring 10 to 30. Five species of fish
3 or -1 years old were analyzed for arsenic: highest con-
centration found in the fish flesh was 0.07 in yellow
perch Highest concentration found
in the organs was 0.27 in skin and scales of bluegills.
?'ater samples from the outlet were analyzed for it)
weeks Starting in _Iune and contained from 0.075 to
0.225 of arsenic.

Ball used ?As-tagged sodium arsenite in a Study
of its effects on pond ecology. Treatment was at 8 ppm.
the standard commercial rate. The sodium at'senite was
taken up rapidly by tlte plants and recycled to the water
and soil by decay and sedimentation of the plants within
5 days. One alga (.S'pimgvm sp.) was killed whereas
another one {Chum sp.) absorbed the herbicide and
appeared to be unalleaed. The tagged material per-
sisted in the water for the duration of the experiment

about one-half the concentration of the applied

nount. The invertebrates (nticrocrustacea) showed great

ariation from very sensiziv to very real tan: in response
no the herbicide. The arsenite nioyetl into the soil and
reached a depth of 5 inches the hydrOsoil during a
60-day period. General eflect on the contmuuitv metab-
olisnt of the pond as shown by the diurnal curve
indicates that the herbicide had a drastic effect. It raises
serious questions as to the advisability of using sodium
arsenite in fish-producing ponds due to its deleterious
effects on several levels of the fish food chain.

(-15) studied the use of concen-
trations of sodium arsenite on bluegills antl pond in-
vertebrates. .-\pplications that mtaled -l reduced the
survival and growth of the fish: immature ?sh more
affected than adults. Substantial quantities of arsenic
were found in the water, attd the organs and
flesh of bluegills at the end of the experiment. Bottom
fauna and plankton populations were reduced or in-
hibited in several pools with the highest concentrations
of herbicide.

Lawrence (Tl) and and .\leehean (125) botlt
used 4 arsenic as a herbicide for filamentous algae
and found it to be toxic to desirable food organisms.
Lawrence also noted that it decreased bluegill produc-
tton.

Amitrole .\larston al. (31) made
a study in Oregon on a stream draining a municipal
watershed. air application was made at 2 lb to
ntrol the salmonberty (Habits l?urslt't. 
ximum concentration of lift was found in water
.tnples 3U mitt later. decreasing and none 6 days. 1.8 ntiles below the

Volume 13, Issue 6 (November), l970

EFFECTS ON 

sprayed area, no amitrole was detected at anv time. Water
sampled during their study probably would ban: been
safe for use in home irrigation syStetns. There was not
sutlicient amitrole present at any time to cause toxicitv
to watm-blooded animals. 

In Other tests with atnit?role for salmonberry control.
Tarrant and Norris made aerial applications 2
successive years using 2 lb lit 10 gal of water. Level
of amitrole itt samples of stream water at the spray
site was it)? in satnples taken 5 min after spray-inc}:
10 hr later. it had decreased IOU-fold to -l pph. Ami-
trole was n0t detected after 3 days. Norris ct al. (93)
also treated 260 acres with amitrole at 2 lb A. After-
wards, stream water contained 42 and 45 of ami-
trole which totally disappeared in 43 hr.

[Jalapon t2,2-dicltloropropionic acid). It is generally
recognized that dalapon ltas a vety low degree 01 tox-
icity to fish. It caused zero mortality to goldfish tCm'ns--
sitts atu'atus after 2i hr of exposure at NO (62).
Thiegs (I30) and .\lagee and Colmer (79) showed its
rapid decomposition itt soil.

Chancellor and Ripper (22 indicated that yellow
perch, nortltent fathead minnow promefas
R.). and pumpkinseed (Lepmnis gtbbosns L.) tolerated
30? ppm, although at 400 of the brown trout
were killed. The sodium salt of dalapon apparently did
not alleCt the Lake Emerald shitter atlterinoidcs
R.) at ppm- Dragon lly sp.) were
not killed by and several insects that normal-
ly are eaten by fish were unafleCted by dosages between
to SUU ppm.

jacorossi in investigations on controlling wech
in canals reported no toxic ellects on seven fish species
for 3 days to dalapon at concentrations lar ex-
ceeding these likely to be obtained in field practice.
Kochkin (tilt), in an organoleptic sutdy ol' dalapon.
concluded that 2 is the maximum con-
centration.

Dic/tlobcnif Frank ct tJ. (ll)
investigated the elfects of canal-bottom plots treated with
dichlohenil and acid ifenacl
on irrigation water flowing over them. Concentrations
of dichlobenil and fenac initially were 5.6 and 3.8
ppm. respeCtively. The bulk of the herbicides lost front
the plots due to the water llow occurred quickly and
safe levels were reached after I hr of water How.

\"an \'alin ?39) applied dichlobenil at Ill. 20. and
id to ponds. Residues in both water and fish were
highest within 3 dva after treatment. Concentration in

'water ll days after treatment was about of that

after 3 days. The residue itt ?sh, whole body, dropped
nearly as fast but could still be detected after 112 days.
Taylor (123. 120') reported that the of dichlo-
benil for most species of was l5 to 20 ppm: the
usual recommended rate of 0.5 did not adversely
affect spawning. It had a low degree of toxicity to species
such as sltrintp (Penman and oysters 
:?irginica (ll: plankton tolerated ll} ppm. .\letliau lethal
concentration to the ftesh water crustacean nia sp..
a common loud tltain tnganistn, was appro\imately 1U
ppm. \\'alker (H2) reported that concentrations of [2 to

739

18251

 
22 with a 96-hr exposure period produced 50%
mortality in four warnt water ilsll species.

Cope et (27) reported on pond studies in Oklahoma
ind Colorado- the former site. only of tlte dichlo-
benil applied at 10, and -10 remained in the
water after 11 days; even with 40 pptn. only 1.5 
remained after (30 days. At that time. the same amount
was left in the mud. l?sut fish reproduction was affected
adversely by the 40 ppm. and there were some patho-
logic effects in the ?sh organs. There was no immediate
mortality. but survival after 16 weeks was for the
controls and 2t), 5. and in the Ponds witlt the in-
creasing dosages. Increase in weight was for the
controls but 169. 169. and for the
other treatments mentioned above, presumably owing
to a more abundant food supply.

In Colorado. 0.53 of dichlobenil. resulted in
maximum concentration of antl 0:13 of the
herbicide the water after 3 weeks. After 03 days. it
declined to 0.17 when there was 1.98 in the
mud. ?sh mortality occurred outside the cages al-
though a lack of oxygen killed all the caged ?sh at the
battom.

Diqttat 6,7?c1ihvdrodipvridn pyrazinediitmt
ion and ion (paraquat).
Allen et (3) fouttd that the use of diquat at 2 for
1 hr produced some prophtlacdc effect on adult and
juvenile Chinook salmon W.)

affected by the pernicious disease caused by the fungus

I)crtnocystirh'tmt sp.

Beasley e! (if (101 found a maximum of 7.9 of
?araqaa: a. 1.7 of tiitlttal. in hydrosoiis where the

rois had been treated 4 years previousiv with 11.3 11) A.

'quat and paraquat will persist in for long
periods of time. Blackburn and \?t'eldon (12} reported
that diquat. originally applied at 2.5 ppm, persisted in
water 8 and 11 days. toxicity to ?sh was apparent.
It appeared to have little effect on natural fauna and
plankton.

Coats et investigated the disappearance of di-
quat and paraquat when applied at 0.5 antl 0.1 
in plaStic pools and ponds. l?araquat in plastic ponds in
water alone took 31 weeks to disappear and diquat 111
weeks. in contrast. neither was found in pond water
after 15 days. This work shows the strong sorptive
power of soil for these compounds. Daly et (31) found
that water analysis revealed only a trace of paraquat and
diquat 2-1 after thev had been applied to control
eurasian spicattun L).

Frank et al.? in comparing the residual effect of para-
quat. an ester of acid (EA-D).
dicltlobenil. and fenac in pond water and soils. found
paraquat was the least persistent in water: it was not
detected 12 days after application. But the butoxyetlt-
anol ester of 2.1-D took days to disappear. Dichlo-
benil and fenac persisted for 150 days antl traces of
fenac were still present in the water 1 year after appli-
cation. After 85 days. the top inch of the 
contained 10 paraquat and no Fenac in

?Frank. P. R. D. Comes. and E. B. Hollineslvorth. 1966.
sistence of hetbicit.es in pontied water and soils- \\'eed Sci. Soc.
mer. Abstr. p. 5



SCIENCE

the hydrOsoil dropped front 12.1 at 2-1 days to
0.26 after 160 days; dicltlobenil went front 10.6
initially to 1.3-1 after 120 days.

Later. Frank attd Comes (12) studied herbicide resi-
dues in pond water and hydrosoil. They applied dichlo-
benil at 0.55 and 0:10 ppm. fenac at 1 and 1.56 pptn.
SEA-D at 1.33 ppm. T-oxabicyclo 
carboxylic acid (endothall) at 1 ppm, paraquat at 1.1-1
ppm, and diquat at 11.62 ppm. Fenac and dichlobenil
residues persisted in soil and water for more than 160
days. Low amounts of were found in water for 
days anti in the hydrosoil for 55 days after which it
disappeared. Endothall. paraquat, and diquat were less
persistent and were not found in the water after 2-1. 8.
and -1 days. respecdvely. Paraquat and diquat persisted
at high concentrations in the ltytersoil for 35 and 160
days. respectively.

Gilderltus (40; expOsed ?ngerling and adult bluegills
to diquat at various concentrations and frequencies of
treatment in outdoor pools for ?34 weeks. Numbers
among bottom fauna and plankton were unchanged.
Treated fish did not show any changes that could be
attributetl to diquat. The herbicide was toxic to a fresh-
water crustacean. Cladocera sp.

Crzenda et a1. (50) compared the persistence of diquat
and paraquat. applied at 2.5 in pond water. to
fenac at 1.0 and at'nitrole at 1.0 ppm. Both para-
quat and diquat disappeared from the water in less
than 1 month. Soil sediment in the water increased the
rate of their disappearance: they did not appear to be
desorbed from b0ttom clays. The paraquat and diquat
treatments were repeated with essentially the same re-
sults. Fenac was reduced to 2.1 by 202 days and
amitrole to 0.13 ppm-by 15-1 days after treatment. He
concluded that fenac appears to have the least potential
for use in sources of pOtable water because of its long
persistence.

Rodriguez-Kahuna et al. (117) studied the effect of
paraquat on the growth of the pathogenic fungus.
Sclerotitun rolfsii Sacc.. in liquid culture and in soil.
In the fonner, growth was decreased at all concentra-
tions. in soil experiments. evolution of was lowered
at all concentrations but greatly reduced at only the
0.5 and 1 tates. Yeo (150) studied the effects
of diquat and paraquat in water on fish. in resen'oirs.
diquat applied at 10011 dissipated to 9 after 12
days. In ?sh-growing pools. diquat at 41100 dissipated
to 900 or less. Paraquat was slower to disappear

and exhibited unpredictable fluctuations.

Beasley et al. (10). using radioactive diquat. found
that goldfish placed into water containing 8 diquat
for attd then into fresh water for 10 days ltad
ntdioactivitv in Abe flesh and various organs in a negli-
gible atnouitt except in the gastrointestinal tract. Cil-
investigated the effects of diquat on blue-
gills and their food organisms. He reported that diquat
disappeared front the water at about the same rate dur-
ing spring or summer. Residues were highest about 10
davs after treatment and disappeared to 12 weeks
after treatment. Diquat at 1 and 3 did nOt affect
adult or ?llgerlittg bluegills. hematological or histo-
pathological effens were noted. It was quite to the
freshwater crustacean. Cladocem sp.. but these returned

Volume 18. Issue 6 (November). 1970

1132:





co
i?v

as
00

wu?v?- .

-- -

52.

 HERBICIDE EFFECTS AQUATIC ORCAXISMS

to their normal numbers with the disappearance of di-
?uat.

Holt (50) used paraquat for aquatic weed control in
ditches. He reported that fish. frogs (Rmm and
aquatic insects appeared to be unhanned by paraquat.
Cattle drinking the ditch water after treatment {how
long not specified) showed no toxic effect. Riemer (llti')
used tliquat at 0.31. 0.53. and 0.73 to control
bladderwort sp.l. No apparent effects were
mated on large month bass fry (.lIicroptei-ns 
Lacepede). pickerel fry rEsox or young yellow perch:
it tlid not affect the spawning of sun?sh.
Bottom dwelling invertebrates. primarily chironomitls.
were unaffected. Plankton were present in an insuf?cient
amount to make valid comparisons before and after
treatment. .

Blok using paraquat and diqttat at 0.4 to 0.8
to control ?lamentous algae. now! no effect on
?sh or fish food organisms. Used at 0.3 for control
of ?lamentous algae. 
urea (diuron) hatl no deleterious effects on ?sh: in shal-
low ditches. no ?sh were killed. He also acted that one
species of rotifiers was present in large numbers and
that one species of Dali/min increased in numbers. The
phytoplankton were chiefly unicellular flagellates and
l?olvox ain't-us Ehr. Dichlobenil antl 2.6-dichlorothio-
benzamide (hereinafter referred to as chlorthiamidf at
were equally well tolerated by the fauna. in
another trial with them. Crustaceae. especially cOpepods
and bottom fauna. increased in number. but the phyto-
ilankton were restricted to flagellates and isolated dia-
nms. Cassie used tliquat at an estimated 0.5 in
New Zealand antl noted a decreased number of diatoms
and algae. The decrease in algae might have been as-
sociated with anaerobic conditions caused by the de-
composing aquatic weeds.

Lam'euce (74) gave a numerical and graphic presen-
tation of aquatic weed data on nine different combina-
tions of treatments Over a r3-year period. He used
paraquat. tliquat. fenac. 
triazine (simarinel. and 
l-methylurea (linuron) in various combinations. This
reviewer interpreted the data as showing that plankton
were unaffected by these treatments with the exception
of the paraquat-linuron combination. This caused a
retluction in plankton the ?rst year and a slight
reduction the following year. Fish production appeared
to have been adversely affected in all treatments with
perhaps the exception of the fenac treatment at the end
of the ?rst year.

Newman (37) gave data on the use of diquat and
paraquat as it affected population changes in the small-
er arthropods. There was a two to three-fold increase in
earthworms: a residual amount of the herbicide was
found in them which was rapidly 10st upon their trans-
fer to untreated soil. .\mounts found even following

high application rates were negligible. in aquatic situa-

?ons no drastic effects on any group of animals t'insects.

mites. wonns. leeches. and flat
-.'ortus') were found. He :59} also reported on the eco-
logical effect of paraquat and tliquat at 0.5 to 1.0 

Volume 18. Issue 6 (November), 1970

at 10 different sites. direct toxic effects to fish were
noted and in one case an increase in Dipterons larvae.
mainly (.?hiromedae. occurred which was associated with
the breakdown of killed weeds.

Newman and \?t'ay (85) reported no direct effeCts of
tliquat and paraquat on aquatic invertebrates present
in their experiments. But they noted severe oxygen de-
pletion at one location following decay of the .treated
aquatic weeds. This resulted in some mortality among
Hirudinea. lsopoda. Odonata. Coleoptera. 'l'richoptera.
Gastropoda. Lambellebranchiata. and captive trout al-
though free-living coarse fish and tr0tit were generally
unaffected. These herbicides were absorbed by the vege:
tation and the bottom mud which in one case con-
tained 3th}. of the original dose in the top 2 inches
5% months after treatment. They also reported on effects
of paraquat and tliquat at 0.5 to at 10 different
sites. No toxic effects to fish were noted. An increase in
dipterous latwae. mainly Chironontidae. associated with
the breakdown of dead weeds occurred once.

Pierce cf of. (109) used fenac. sintazine. 2-chloro--l-
fatrazinc). 
(prome-
tryne). diquat. antl paraquat for aquatic weed control.
The triazines were effective for control of filamentons
algae. Microcystic blooms were removed and rapidly re-
placed with more desirable plankton algae when atra/ine
was used at H.U3 and 0.16 ppm. Diquat and paraquat
used at 0.25 to if) had no adverse effect on fish.
?sh food organisms. or plankton.

Tatum and Blackburn used tliquat at 0.5 
in two sub-tropical ponds. Phytoplankton was tempt".-
rarily atlet'teti nntnetliately but recovered rapidly. Ul-
ganic matter from decaying vegetation appeared to
significantly increase certain benthic organisms.

Wile (HS) Studied the effect of tliquat. fenac. and situa-
zine on phytoplankton. .\pplications were at :md 13
In two ponds where diatoms were dominant oyer
the flagellates and in two where blue green algae were
dominant. there was no reduction. There was a suff-
stantial reduction in one pond wltith had a
distinct bloom at the time of spraying. in agreement
with Hilsenhoff (55). no effects were found on bottom-
dwelling organisms examined in dredge samples. 
tliquat was detected in the water 5 and 7 days later.
was used at to 3 ppm. In three ponds. tllt't'e
was no effect on the phytoplankton. In three Others.
the numbers dropped. but total recovery was observed
in ti to weeks. In one pond. the population was re-
duced 75?; and remained there for the rest of the sea-
son. effect was noted on bottom-dwelling organisms.
Simazine was applied at 0.5 to 3.0 to control
aquatic weeds. but phytOplankton studies were not made.
it has potential in control of filamentous algae.

Endof/tafl. Liutlaberrv (75) reported that although the
normal dosage of endothall is to 2 ppm. 5 endo-
thall completely disappears in ?l days. He also reported
no residue itt fish which were placed in water con-
taining ti and 12 ppm. Bluegills tolerated. with no
mortality. ll'fl for Lil days. There was no mortality
in three species of minnows sp.) at -lH
ppm. Salmon (.S'rtfmo rainbow trout gainf-

74!
on

Moo

6f7dtff79

teasa

 SCIENCE

ncrii R.), and bass (.lficropfcrits 5p.) suffered no mortal-
ity at ppm.

Simes (120) applied endothall at 5 in ponds or
trout-rearing raceways in a fish hatchery
and acted no indication of distress by the ?sh. Steuckc
(123, l24) reported that 2 of in fish
hatchery pontls did not affect the ?sh pepulation?large
mouth bass, bluegills. goldfish, or ?sh food organisms.
Walker (141) reported that it took 25 days for disappear-
ance of residue from ?sh flesh expOsecl to to 3 
of endOthall; some tesidues were found in ?sh food or-
ganisms 3 weeks later.

Fenac. Pierce (104, 105, applied fenac to ponds at
to 3 ppm. There was no eflect on plankton, small
invertebrates, frogs, turtles (Cliclona and fish. Young
?sh were present.

Paraquat. See section on diquat.

Phenoxynliplmtic acids in renter. A study of the effect of
in irrigation water on red Mexican beans i'P/iasco-
lus vulgaris L.) has been reported by Bruns When
the herbicide at 2 in the water was applied to beans
in the seedling stage, root systems were severely in-
jured; recovery began within 16 days. When the dosage
was increased to ti yield was reduced 40%. The
2 lb-rate, when applied in the bloom stage, caused no
signi?cant reduction in yield; but the loss at 6 
was 

Aldhous (2) used the nonyl ester of 2.4-D at 4 lhfae 1?2
gal water to treat a forest of Sitka spruce tPicca sitciicnsis
(Bong) Carr.) for control of sc0tch heather (Gallium
vulgaris Salish). Water samples, from a ditch that
drained about 3 acres. were taken before spraying and
up to 4 weeks. .\nalysis showed 0.0 after 28 days.
Aly and Faust (4) found that 2,-l-D disappeared in 35
days from lake bottom soil that had been previously
treated: on untreated it took 65 tlays.

Averitt (6) in one ?eld test applied an amine formu-
lation of 2,-l-D to water hyacinth IEfch/cornfa crassipes
Solms). -l lb 10 were found at
3 weeks and 0.6 at 2f) weeks. In anodier ?eld test,
when 4 lb 2,-l-D were into the water at two
different locations, he found 689 and 967 day
atter application: 31 days later. he found If and 19
ppb. In other tests. when the original applications were
-l and 5 he found -l and I5 after 39 days.
Averitt5 also applied the methyl amine salt of at
?1 He found 802 the second day, HG the third.
the fourth, and then a gradual decline until .102
days when no 2.4-0 was detected. Similar results were
produced by acid
(silvex) and a polypropylene glycol butyl ether ester of


Bailey at of.? applied to ponds the polypropylene
glycol ether ester of silvex at 8 Half of
the ester was hydrolyred to the acid hr. There was absorption of both acid

5.h-eritt, W. K. 1963. A summary of a studv of the persistencv
and residues of some herbicides in surface waters. Weed Sci. Soc.
.\mer. Abstr. p. 63.

?Bailev, G. W.. I. D. Pope. Ir" and D. R. Cochrane. 1963. The
degradation kinetics and of silscx under impounded

inditions. Weed Sci. Soc. of Amer. .\bstr. p. 43.

742

and ester upon the houom sediments with complete dis-
sipation of both alter 5 weeks. Cochrane ct al. (24) ap-
plied b' lbl'A of the same ester of silvex to three plastic
ponds, each with a different soil type at the bottom.
in 2 weeks the ester hydrolyzed totally to the
acid which gradually dissipated met 19 weeks. Absorp-
tion of both ester and acid apparently occurred on the
followed by gradual diminution. There is a
passibility that silvex acid and/or a degradation product
may be desorhed and re-enter the water. Therefore,
they concluded that it should not be applied directly to
water supplies for human consumption.

There have been few reports dealing with the effects
of herbicides applied to watersheds and reservoirs where
the water is destined for human consumption. One 
Government publication (134) lists 0.1 for LIA-D,
and siliex as permissible criteria for surface
water for public water supplies. such criteria are

given for Other herbicides. The same reference has a,

very useful table giving the likely concentration of
herbicides in irrigation water resulting after a typical
application rate and also the crop injury threshold in
ppm.

FauSt et (35. 37) investigated drinking water qual-
ity as affected by and 2,4-dichlorophenol (herein-
after releiied to as The latter is a most p0tcnt
oflender in water as it causes an objeCtionable medicinal
taste at 8 and has an odor even at 2 ppb. ln lab-
oraton carhoi tests using 9.5 and lti.7 of
were released in 7 days. .\laximum concentra-
tions of dexeloped in 218 days. When the car-
hoys were aerated and in the presence of small amounts
of oigauit matter. this phenol disappeared rapidly. Ct.-
tlc-r acul tonditions antl water unfavorable for biological
Oxidation. about half of the persisted.

Faust e! (If. (35) in later ?eld tests applied the isooctyl
ester of 2.4-1) which contained 2,-l-DCl? as an impurity. it
was applied at ?20 lbl-?A to control eurasian watermil-
foil. 'llus east: a theoretical concentration in the lake
of lfil anti 4.36 of This did
nOt tame odor. There were 13 of al-
thouqu ll did not appear to accumulate as the result of
biolueual degradation of Eel-D. .\faximunt level of 
was i?J 3 ppb. dropping to 13 after 132 days.

Frank and Comes introduced into a pond 1.33
of from a 20"; granular formulation. Initial
was 0.02! ppm. The maximum. 0.067
ppm. (K'Hll'lt?tl after 13 days and fell to 0.019 
after '14 This formulation persiSted longer in the
lmlmunl but could net be detected there after 55 days.
Clihanm .fh?i reported on the contamination of water
in a rest-non with the butyl ester of which
rendered it un?t for use by organoleptic criteria. No
cases of poisoning were reported.

Gucnda in a public health report on weed con-
trol in potable water supplies. gave data concerningsix?
common herbicides and their persistence in water after
vanling periods of time. Application was a maximum
of -l ppm. .\fter some days. no more than 1 was
fomnl. For the dintethyl amine salts of 62 days
after treatment. the amount had dropped to 0.00] ppm.

Cratekowski (if) and Tarrant and Norris did
aerial forest spraying with a commercial 50-50 formu-

Volume 18, Issue 6 (November). 1970

M00



. -w?piml.







?ation or low volatile esters of 2.4-1) and 2.-l.5-T diluted
.?ith diesel oil and applied at 2 lb A. This gave resi-
es of to 70 in the water. [isuallv tlte level
.ell'in a few days to 0.2 ppb. Maximum time for total
disappearance of 2.l-D was 2 days at one test sration and
17 days at another.

Krammes and \t'ellets :70) sprayed a brush-covered
area on a steep slope to encourage consersion to grass.
Application rate was lb 2.-i-D and ?it lb A 2.4.5-T
as low volatile esters in a 2tt-gal mixture of water plus
gal of diesel oil. A follow-up hand treatment of brush
was made with 1.3 gal of 2 lb each 2.4-D and 2.-l.5-T If!?
gal of water that contained 1 gal of diesel oil. herbi-
?cide was found in surface water. In soil samples. small
of herbicide were detected 8 days later. .\?one
was found after months.

Noll and lint-dick t92) applied to a lake an ester of
silvex at 2 ppm. \\?lten heated before treatment. the
water ltad an earthv. musty Otlut'. After treatment. a
kerosene-like odor developed. This odor lessened con-
siderably in 1 week although it was still deteCIable 3
weeks after treatment. The lake was treated in two sec-
tions; :1 number of large and small ?sh were killed
when the second half was treated although this was not
noted for the ?rst half. A second lake was treated at
a concentration of and no fish were killed.

Norris (-1 (if. {93) applied by helicopter a phenoxy
killer to 4713 A at a rate of ll) of
2.4-D and lb of 2.1.5-T ll) gal of oil A. Anather area
. was sp0t-treated in 23 locations resulting in it)? treated

Tea. A stream drained each area. Samples taken from

)(ll streams were analvzed. Measurable quantities of

rbicide were nor. in am samples althOugn a
"mud of 3H: to 51-1: inches of rain had fallen over the
treated area. Norris (El-ll cites ivork by Linden and
Muller (Tti; who used diesel oil at rates in excess of
500 gpa. which was followed bv leaching with 4 in

'of'rain water. A maximum of 2 of diesel oil was? 

found in a sandv loant soil at a depth below 215 inches.
It was concluded that application of (llC'\Cl oil at rates
of 5H gpa to soil surfaces presented no threat to ground
water quality.

Reigner ct n1. (ll-l) discuss treatment of vegetation
along two stream banks with the usual amounts of acid
and ester formulations of Water samples were
taken for :tnaltsis periodicallv for 3 weeks. Odor tests
were conducted. Tltev concluded that 2.45-T. if applied
with normal precautions. could be used on municipal
watersheds without creating any water contamination.

Reinhart (H5) applied I325 gal (Tiff) gal of diesel oil
plus 52 gal of an ester of containing 6 lb gal} on
30 A of timber watershed. This was a basal spray
applied to all trees 1 inch or larger. This treatment
is heavier than would be used in a riparian area and
diesel oil normallv would not be used. An odor panel
found no contamination in numerous water samples
taken front the stream draining the treated area.
Schwartz (119) reported a laboratorv of aqueous
solutions of 2.-l-D under conditions favorable for mi-

'robiological activity. At least tilt"; of the 2.4-D persisted

?3 to ti months.

.nnith and lsotn applied a 20?; formulation of

butoxy ethanol ester of 2.l?D to control turasian

Volume 18. Issue 6 (November). 1970

EFFECTS AQUATIC ORCANISMS

watennilfoil. Application of 83.5 tons of tile 2.-l-D was
made to infested acres in seven reservoirs
at rates of -lt  to It"! lb A. Analyses showed little up-
take of 2.-i-D by ?sh but some by mussels {My-trim 
all mud samples showed some Eight of nine water
treatment plants showed less than 1 of 2,-l-D in the
water. The ninth had its water intake direcdy under
a 2.4-0 application plot and its highest concentration
was I and 2 ppb. Thev also reported that mosquito
larvae (Anopltries (IHatfi?i?tflfuf?fNS L.) in the laboratory
survived 2.4-D treatments at ppm: adults from these
larvae were carried to the difference in hardi-
ness or reproductive ability could be detected. They

concluded that high rates of 2.4-0 applied in TVA-

reservoirs have not produced adverse effeCts upon aquatic
fauna or water qualitv.

Wershaw er (HT: reported on a literature review
of an ionization constant study of Various authors
showed 2.-l-D could persist for periods up to (3 months
in natural waters. \\'inston and Ritty (HQ) discussed
the fate of phenoxy herbicides when applied to a water-
shed. At least eight species of soil bacteria use phenoxy-
aliphatic herbicides as an energy source. Thev concluded
that the phenoxvalipitatic herbicides are decomposed
into carbon dioxide. inorganic chloride ions. and water.
and when properlv appiied to watershed areas will not
create a water pollution problem.

acids and ?sh. Beaven et :11. (Hi had
caged ovsters. blue crab; .snpidns 

clams ,t?fvn and fish near the center antl out-

side of titlewater plots set up to control Eurasian wa-
tet'milfoil. The ester of 2.1-0 was used at.
St). an. and 15:) lb The two latter treatments were rep-
licated three times. When normal aerobic conditions
were maintained. treatments at 30 resulted in no
mortalitv to the native microfanna and the valuable
native plants. 

Butler (20) exposed continuously for 7 days to
?owing sea water containing of the 
ethanol ester of mortalitv occurred. The oysters
had accumulated l8 2.4-1) acid. Half the treated
oysters then were flushed with normal sea water for
7 days at which time they ltatl no 2,-l-D residue. In
Other experiments no etfects of were noted on
pigtoe mussels (Pit-urm'u-na corrinlum Creeul. Data are
given for effects of 37 different herbicides on ovstets.
shrimp. fish. and He concluded that
herbicides in general are less toxic than most Other

pesticides.

Huehes and Hughes and Davis (?ll reported on
variations of different formulations of 2.4.5-T.
silvex. and acid (cli-
chloroprop) to bluegills. The number of different for-
mulations varied from three with tlichlorprop to 21
for 2.4-D. Differences in toxicity of various formulations
of the same herbicide were great and ntav be greater
than the difference in toxicity between different herbi-
cides. Amine salts were generally less toxic than esters.
The actual formulation must be evaluated to determine
the toxicity.

Lawrence ct it]. t72?. ran experiments for aquatic weed
control with 2.-l-D acid and silvex both at 5 ppm.

7-43

asses

on



 SCIENCE

I .
diquat at ?3 ppnt. diquat at plus 3 {t-l-chloro-
o-tolyl) oxyt acetic acid endothall at 2 ppm.
and antitrole at L5 3 They placed
fatheatl minnows and redear sunfish micro-
loplms G.) in their plastic pools. deaths up to 9
months after treatment occurred with either fish: in
several instances thev had Mackenthun (73)
noted in using ill-D granules to control aquatic weeds
in ?sh hatchery ponds that applications harmful effects. Rao and .\lurty
(112) found that using the sodium salt water to control [point-a L.
caused no fish mortality.

Rawls (HS) worked with blue crabs. eastern oysters.
soft shell clams. and various fish species in conjunc-
tion with herbicidal experiments. The following herbi-
cides were applied at these rates as lb 2,l-l)
acetamide?QO: polypropylene glycol butyl ether esters
of silvex?l?: butoxy ethanol ester of ill-D
-?20 to 120: and isooctyl ester 0! to (EU. the
end of month. he concluded that only the 2.4-D
acetamide appeared dangerously toxic.

ThOmas and Dulfy investigated both in labora-
tory and ?eld the effect of the butoxyethanol ester of
for control of eelgrass (Zostera marina L). They
qu0te Mount and Stephen (84) who stated that con-
tinuous exposure to 0.3 Elsi-D had no effect on
reproduction of fatheatl minnows. Rawls (113) also is
quoted as saying that ill-D had little etfect on
oyster egg development: there is evidence that Lil-D
residues in shell fish are eliminated in l? mouths. Butler
(18. IO) also indicates that Continuous expo-aire- to 3.7-3
2.41) reduced oyster shell growth by :?illnj: 
for a -l-hr exposure period reduced phytoplankton pro-
ductivity by In their own laboratory experiments.
Thomas and Duffy found that with application of hi
the highest concentration of detected in the
water was 0.l ppm: the lilsl. detectable amount Occurred
20 days after treatment. In the field experiments at 30
and in water samples taken for consecutive weeks.
no was detected after 2! days. Highest field con-
centration was l.-l ppm. In the laboratory experiment.
highest residue detected in oysters was 3.07 ppm:
no 2,-l-D residue was detected 17 days after its disap-
pearance from the water. llc-nthos samples from the ?eld
showed generally increasing fauna biomass during -l
months following treatment: t0tal number of animals
also rose. mortality of natural fauna or ?ora was
observed and relative percentage of species remained un-
changed.

Tschirley (133) discussed the effects in Vietnam of
using both phenoxyaliphatic weed killers and -l-:unino-
acid (piclorant) applied over more
extensive areas and at higher dosages than nonnally
used on crops. Statistics on the ?sh caught over a 3-year
period. including fresh antl salt water fish and also cuttle-
?sh (Sr-pin sp.?t. mollusks. shrimp. and crabs. indicated
that the aquatic food chain had not been seriously dis-
turbed.

Estes (31) applied to polypropylene glvcol ether
esters of silsex at 3 lb to control aquatic weeds. Within
30 ntin after spraying where the average depth of water
was 3 ft. some fish seemed distressed attd some pitkerel

744

(Box sp.; died. lit a setonrl area. no distressed fish were
observed. Analyses of ?sh after the treatments showed
less than (Ltll silvex: there was no change in the
population. Cavlor and Home!? reported
on 3 years work with silvex used lor aquatic weed cott-
trol. Results are given for eight species of aquatic weeds

with good results for three of the most troublesome. Xoc

dead ?sh were observed. although the ?sh hunted new

habitats when their old plant coyer was destroyed. 

Homer and Caylor (53) used endodtall and silvex
for control of water weeds. These herbicides caused no
?sh mortality, nor were there reports of off-flavor in the
?sh caught by many anglers in that area. Houser 
reports that three species of ?sh in two ponds treated
with silvex experienced a weight loss in the 2 'years
following application. It is net known whether this
was direttly due to the silvex or indirecdv through the
destruction of the weed beds. Mathews reported
that lit New Zealand. trout (Sultan sp.) were ttnalleCted
by 5ft silvex.

Mullison (85) reviewed the silvex literature and gave
toxicological data for rats. guinea pigs (Cat-in porcellus
Ll, rabbits. mice. chickens (Callus gull?: L), dogs (Curtis
familim'is L). and two species of ?sh. Silyex apparently
does nOt constitute a hazard to game birds such as
bobwhite quail (Cofinus virgininnus mallard ducks
{Aims pints-rln-nrho LI). and ring-neck pheasants t'P/mnu-
colc/n'cas'LJL There seemed to'be fish species varia-
tion in regard to tolerance of ester formulations of silvex.
but in mast situations where a silvex ester was used.
?sh populations were not seriously affected. Nonethe-
less. the potassittnt salt of silvex was less toxic to fish
than the ester of silvex, Effects. of the ester of sil're?: on
plankton were variable: some injury was observed in
fresh water. this was temporary attd populations
soon were back to normal.

acids and plankton. Butler FC-
ported physiological data-on decrease in carbon 
of a natural salt water phytoplankton community. chieilx
dittollagellates and diatoms. It was given a -l-hr expo-
sure to 1 of various herbitidal formulations. There
was a wide variation in response even among members
of the same group of herbicides such as dilfcrent esters
of Ell-D. The san'u. paper contains muth 1-201?? data
on effects of herbicides on oysters. shrimp. and fish.

Cowell mod silvex at concentrations of in a
study on aquatic vegetation and plankton in farm ponds.
It gave excellent control of I?otnmogct?mt sp. and Lemon
sp. but not satist'acmry control of the alga. Claim sp.
There were no adverse effects on the phytoplankton and
rooplanktou: no ?sh mortality was observed. Cowell 
reported that neither silvex nor sodium arsenite. at the
dosages used. had any effect on plntoplankton: ?3 
of silvex were nOt too toxic to zooplaukton. but sodium
arsenite at ?l drastically reduced zooplankton and
heasilv decreased reti?ers. copepods. and ?cladocerans.
Silvex'treatments had no such effects.

(lrosbv and Tucker studied the effect of various
herbicides on 1.. after a ?lti-hr exposure.
They reported ?3.l-D and were innoatous. but

concentrate which causcs response in of the
indiy tdttals.

Volume 13. Issue 6 (November). 1970

18235

It



WA

6



(hereinafter referred to
as (licltlotte). S-ethyl 

3_ (molinate). (propanil).
. .td dichlobenil could present a danger to Dupltnitt un-
der, ?eld conditions if used at excessive rates.

De Oliyeira (33) reported that a liracilian lake sprayed
witlt an amine of 2.l-D temporarily increased in plank-
ton. This was attributed to a better food supply caused
by tlte dying weeds. Harp and Campbell (it-l) 
the effects of the porassium salt of silyex on benthos
?organisms of a farm pond. It was applied at 2.8 to ~l.ti
and tire pond was sampled at 23-day intervals for
3 months. The benthos population doubled. There was
"an increase itt tendipetlids. aligochaetes. and chaoborus
correlated with an increased food supply from the de-
caying plants. Chaoborus and libellulids increased greatly
within 1 year after treatment. Snails. leeches {Hit-mic
and coenagrionids appeared unalfectetl. Lacewing
flies sp.) decreased quickly but reappeared in
smaller numbers the last 2 months.

Petruk (97) applied 2.-l-D and trichloroacetic acid
(TCA) on saprophytic microorganisms in ponds. The
second or third day after the sodium salt of was
administered. the population of microorganisms in the
pond increased about 509;. but it gradually diminished
after 12 to days and approached its original value
by 25 days. Similar results occurred with TCA. Varia?
tions of the microorganism population. particularly with
2.4-D, strongly suggested that the herbicides were being
used as food.

Pierce (93. 99, 100) has studied in depth the poly-

ropylene gylcol ether esters of silt-ex. Plankton
as represented by about 50 species in the following
asses. Annelida. Chlorophyceae. (Irustacea. Desmida-
ceae. Dictomaceae. Flagellata. Myxophyceae. Nematoda.
Rotifera, attd other protozoa. llentltic organisms were
represented by the following groups: Amphipoda. Anne-

lida, Diptera. prertterida. Castropoda. l?elecypoda. and 

Odonata. Also present were large aquatic vertebrates.
such as fish. frogs. and Silt-ex at 1.2 tempo-
rarily decreased the plankton population. it was
back to normal within 2 weeks. The bentltic organisms.
?sh and other larger aquatic vertebrates were ttnalfected.
Pierce compared two formulations of the potassium
salt of silvex at 2.2 attd observed no effects on plan k-
ton or bentltic organisms. Fish of all ages, frogs, and
turtles were present throughout the summer attd were
unaffected. Pierce 102) made anodter series of in-
vestigations using granular 2.4-0 at rates of 1.5 to 6.2
ppm. effect on the bentltic or plankton population
was noted. although in one trial there might have been a
temporary reduction in plankton which to
normal in 3 weeks.

Pierce (107) determined the effects of a number of
herbicides on plankton attd other pond organisms. Di-
chlobenil. fenac. acid (di-
cantba). endothall. the potassium salt of silvex. attd 2.4-D
acid in a granular fonnulation were used at dosages of
to 2.5 ppm. There were no significant changes itt the
number of individuals or species in the planktott.
\'either the bentltic organisms. consisting chiellv of chiro-
tnid lanae and gantntarids. nor any other of the pond
:llers -ltowed any injury. Pierce continued her

Volume 18, issue 6 (November). 1970

EFFECTS ON 

ecological investigations using a combination of 
attd ettdothall ting} to give a concentration
of 2 ppm. This ltad tto deleterious etleCts on plankton,
large and small ?sh. tadpoles. frogs. turtles. and water
snakes 

Walker (l-ll'll reported that bottom-dwelling organisms
in ?sh ponds were significantly reduced in number by
tnebtni-
on), sodium arscnite. and copper sulfate dttring the
year following treatment but were not allected by en-
dothall and sintazine. Dosages were net Stated except
as "reasonable concentrations". He discussed the possi-
bility of sublethal dosages changing the species composi-
tion and how that would affect pond ecology.

ll'hitney? studied the effects on aquatic fauna of the
polypropylene glycol butyl ether ester of silvex applied
at Fish deaths were observed in half the
silvexapplications: giuard shad spa. blue-
gills, attd largemouth bass were the principal species
affected. Highest value found was l9 2-1 hr after
treatment a forage This decreased to a negligible
amount I to 3 weeks later. Benthic organisms increased
signi?cantly itt number after treatment. The normal
phytoplankton consistincz of numerous species was de-
strOvecl and replaced by sp. attd dinoflagellates.
It did n0t return in 3 weeks. Rotifers and cruStacea
also decreased following application but regained their
normal ntunbers within to 3 weeks.

Piclomtn acid). Davis
et? (32). after using picloram at an unusually high rate
(9.3 a pellet on a watershed area.
found detectable of piclorant in the stream after
rains. Highest concentration found in the stream was 0.37
ppm. After 16 months. during which a total of 40 inches
of rain fell. no piclorant was detected in the stream.

Butler using a formulation of piclorant and 
at 1 ppm. reported no effect on oysters. shrimp. fish.
and phytoplankton. Kenaga studied and reviewed
the effect of piclorant on if; ?sh species and three game
birds. This work indicated that it presents an extremely
low pOtential haiard. if any. to such wildlife.

Hardy (53) showed that piclorant at 1 did not
retard the growth of algae or Guppies were
grown in water with piclorant and fed 
reared in suclt treated water. The guppies were normal
in appearance. behavior. and reproduction. The Drip/mitt
also grew and reproduced nonnallv with no evidence of
biological magnification of the piclorant in them.

Sorfitmt PCP (sodium Isltizawa and
Matsuguchi (Ft-l) reported that the sodium salt of PCP
mixed with soil at a rate of stimulated bac-
terial growth. Pst'ttrlontoims sp. becoming predominant.
.-\mmoniftcation and iron retluction were depressed at
concentrations greater than [?10 ppm.

The Okubos (95) reported that an exrensive nuntber
of fish and clams were killed in IBM and 196?. ?on the
coast of the .\riake Sea. Kyushu. PCP applica-
tion had been followed at once by a heavy raitt. They
concluded that the resistance of a native clam l'cnerttpis

?\\'hitney. E. W. was. The toxicity of silvex to aquatic fauna.
xoc. of All!" Abstr. p. 

745



?on

is 
b.

SEMICS



lAf?a?



 1)

sp.) to PCP decreases to one-tenth its normal value
under stress of the diluted sea water.

Saito e! (HS) used the sodium salt of. PCP as a
359'; granular material applietl at rial on rice
(Or-ya: satit-u paddy ?elds. Samples of river water
were collected at diflerent spors near the treated
?elds. Maximum quantity of PCP detected was 50 ppb:
they concluded there was no danger front this use of
the herbicide.

s-Triazines and 
loluidine Axe et (8) studied the residual
effect in farmers' ?elds of atrazine, 2-chloro--l.6-bisiiso-
propylamino)-s-triazine tpropazinei, and triiluralin. Only
of the tiilluralin was found in the soil after 3
months. propa/ine after 5 weeks. and 33"?o of the
atrazine alter 5 days. Highest concentrations of herbicide
in the runoff water were (Ln-l. 0.05. and [or
atrazine. propa/ine. and trifluralin. respectively. It was
assumed that much of the herbicides must have been on
or in the plants.

Avraman studied the effect of prometryne on the
taste of water and its toxicity to mammals. The herbi-
cide, within its solubilitv limits. did not affect the color
or odor of the water. The taste threshold was 1.2 ppm.
The short-term value for mice (.lfus musculus L.)
was 9.3 to 20.6 gr-lb? and 19.2 to gr lb? for rats
nonmgicus Ll. Daily doses of 4.3 10"4 gr lb"
in long-term studies had no signi?cant eliect on rabbits
(Oxyctol?agus cmticulus L.) and rats. He concluded that
a level of would be the maximum permissible
concentration in water.

and Cihegotskii (3151 investigated simazine.
atra7ine. proparine. 5-amino--l-chloro-B-phenvl-B 
pyridazinone (pyt'azon?i. sodium antl diuron in
water. \Vater containing 0.1 to 5.0 of these herbi-
cides did not tlilfer from control samples in its oceano-
leptic properties even after heating to I58 F. 
excepted. increasing the concentration of these herbi-
cides to 50 to ?20? markedly chanqetl tlte orqano-
leptic properties of the drinking:r water. particularlv for
pyrazon. Dinion excepted. ga'ound water containing; till?
to ol? these herbicides stored in bottles for 3
to 20 months at temperatures of 43 to 71 showed no
drop in concentration. Ground water containing 50 to
of these herbicides was treated with 3m: 2-: ill?3
g?r-?qt of calcium hypochlorite: this completely decon-
taminated the water.

Weldon and Blackburn (H5) reported that E-t'etlnl-
(a -
tryne) persisted in plastic-lined pools nearly unchanged
for 32 days after application. In the ?eld. however. no
residues were found under treated water hyacinth 1 day
after application.

Snow used simazine at 2 in two ponds. In
one pond. adult retlear sun?sh were killed although
no small ?sh died; none died in the other. Anmher
pond was treated at 3 antl no ?sh died. The
phytoplankton developed slowly after 4 to 6 weeks. Fish
production was low to merage. but the evidence was
inconclusive as to the cause-

?'alker experimented with sitnazine and Other

'.\voirtlupois units.

7-16

SCIENCE

s-tri:uines in ?sh habitats. Sima/ine was less toxic than
propazine. 
(prometonei. or atra/ine to fish. Laboratorv tests on bot-
tom oreanisms showed simaziue had an ol ppm:
howeter. ?eld obsenations did "at demonstrate am
serious reproduction problems. .\trazine is more toxic
titan simazine to bottom organisms. but they recovered
in -l to ti months. Prontetone and propazine were not
critically examined. but no ?sh mortality was observed.

Flanagan (39) reported simaiine as non-toxic to ?sh
at 3 ppm. Gttse in a summary of reports on aquatic
weed control. mentioned that ametryne at 4 and 5 
caused irritation or death to some fish. Prowse 
noted that TCA at caused no distress to, ?sh
after several tlays' contact. He also noted that simaziue
at caused mortality in -l hr: after l1?
hr the tanks were no longer toxic. \l'ellborn (ll?i in-
vestigated the ?sh toxicity of diquat. diuron. and situa-
zine and found to be the most toxic to bass
(Roccus sp.) with an of 0.25 ppm.

Tri?rtralin. Cope stated the concentration of
trifltuaiin for a 2-l-hr exposure of 2 to 4-inch rainbow
trout was to antl for smaller blue-gills it
was 23 to l?lO ppb. Increasing the temperature increased
the toxic response: trifluralin once was 130 times
as toxic at 85 as ati-15 F. In some cases. there was
greater toxicity in soft than hard water and in other
cases there was no ditierence. (lope later reported
the values of trilluralin to be to ill [or
rainbow trout and bluegills.

Latn'encelo incorporated trilluralin in soil 7 tlats
before putting it in water-?lled pl.hllL ponds. The int-ids
then were planted with several species ot' water weeds
and stocked with fatheatl minnows. There was no mor-
tality to the ?sh during -l weeks after ?ooding; they
reproduced satisfactorilv.

Parka and \l'ortlt :?Jtil found that trilluralin had an
of 58.2 to bluegills: when put on soil and this
soil added to water. to times more were required
to obtain an in a simulated ?eld test thin: swim-
min: pools. trilluraliu was sprayed at the rate til 1 ll) 
on soil .tnd tilled into it. it then was given in inches
ol simulated rainfall: water draining from the treated
area was not toxic tt) bluegills. Analysis of the sediment
showed 55 This chemical has a water
solubilits of 50 ppb. is strongly absorbed by soil. and
leaches very little even on sandy soils.



urt'ns. \Valkev tH-i?i lound monuron. diuron.
l't-iun'on. neburon. and mixtures of them with 
eliectively controlled certain aquatic weeds. Monuron
autl lezturon were less toxic than diuron and neburon
to ?sh. These urea herbicides mixed with were
somewhat more toxic than the urea herbicides alone.
Species dillerences were manifested. Fineetlines were
more sensitive than adults. Fish food organisms were
noticeava reduced in the treated areas.

Frank 1 studied the disappearance of montu'on and
nebutnn in an aquatic environment. Signi?cant hm did
not ?rst? until after .s weeks for monurun and lti
weeks for nebtn'on, The rate of loss tlten accelerated

"'Lzhn-ncc. I. M. 19043. [ticcts c-f mil applitd truluraiin on ?sh
in mums. Win-E Soc. of .?ttner. .\bstr. pp. 

Volume 15. Issue 6 (November). 

0


f?

p'rnrx

In

MULLISOX

until about of the herbicides disappeared. after
which it continued at a decreasing rate. Alter lif?s' weeks
'pproxitnately l5"; of the monuron and 39?; of the
neburon remained.

de long (l6) used diuron at concentra-
tions up to tt.-l in ?sh spawning ponds: it ditl itOt
inhibit the growth of pike. First and second year carp
suffered sonte growth inhibition attributed to the re-
tarded development of ?sh food organisms. Hambric (fill
used diuron at 2 lb or 0.3 lb foot of water on
two lakes in Texas and no ?sh were killed. Maloney
(80) used tnonuron at concentrations of OJHH to 32.?
to control algae. Toxicity data are given for 32
species of algae with exposure periods of and 28 days.
He also noted that ponds treated at 1 and LG 
were not toxic to ?sh. Pierce (lit); used diuron at 0.25.
0.33. and 0.5 to control common duckweed (Lt-mite
minor and nOted light to moderate ?sh mortality at
all concentrations. He attributed this to a low oxygen
supply antl not to the herbicide. plankton bloom
developed in these ponds within 6 weeks.

Alabaster (I) in a study involving l?-l different chem-
icals. many of them herbicides. pointed out that the
toxicity of formulated pesticides can not be predicted
from knowing the toxicity and percentage of the active
ingredient. lt is essential to test the actual formulation
under consideration because many additives are marked-
ly toxic. some more so than the active pesticidal ingre-
dient. Xishinchi and Hashimoto {'Jll studied three kinds
of fresh water fish and two kinds of fresh water 
and their susceptibility to 76 different pesticides. 
hnugli some herbicides were toxic to the species lC?ttrd.
'.iey conclutlcd most herbicides were non-toxic to both
?sh and Drip/mitt.



- This literature survey presents a very considerable
mass of evidence concerning the fate itt our aquatic
environment of various herbicides. Evidence at hand
shows few signs of either a contamination problem or
an accumulation in our waters if our presently available
weed killers are used according to label directions.

The toxicity of herbicides to ?slt is a complex prob-
lem. There is the ttsual biological variation in size. age.
and species as well as physical and chemical differences
at the aquatic site. There also may be variation with
different fonns of the active ingredient: for example.
the esters of plienoxyaliphatic herbicides are tnore toxic
than sodium or amine salts. Additives to a fomtulation
may be markedly toxic. some more so than the active
pesticide ingredient. Therefore. the speci?c formulation
should be evaluated to determine its LDgn. In addition.
the Ll);n may nor indicate the actual hazard to ?sh
because of the circumstances involving the herbicidal
application. For example. when applied to
the soil is strongly adsorbed and moves little. This cre-
ates no real hazard to fish except for accidental con-
tamination. even though the compound per sc is
extremely toxic to them.

Plankton indicate that our present herbicidCS.

luding those used in aquatic sites. have at most only
temporary harmful etfect.

Volume 18, Issue 6 (November). 1070

HERBICIDE EFFECTS AQUATIC 

There is no evidence of a biological magni?cation
problem with herbicides in the food chain organisms.
the intrinsic fact that herbicides kill
plants which then decay and are not generally eaten
shottld prevent biological at one begin-
ning part of the food chain. the plankton. 

johnson c! til. discussed the studies on pesticide
being carried out cooperatively by several gov.
departments. He found that of the many pesti-
citlal chemicals now in Use the most important to ?sh.
wildlife. and estuaries are DDT. its metabolites, and
other chlorinated hydrocarbon insecucides. a single
herbicide is on his list. Brown and Xisltioka (H) dis-
cussed the results ol their monitoring program. Eleven
streams in the western part of the L'nited States were
checked for nine insecticides and 2.4.51?,
and all\ ex. herbicide residues were found.

Most hetbitides now in common the are low in tox-
icity to man and animals. and they break down in soil
and disappear trom water. in addition they are applied
in such a way that they normally do nor accumulate to
create a hazard in our environment. Nonetheless. com-
placency is not justi?ed. Continued careful study of
each netv herbicide is essential with the express pur-
pOse of preventing environmental contamination in the
future. Man's t0ols frequently interfere with nature.
and licibitides are a powerful tool. It is the wisdom
with which this extremely useful agricultural tool is
used that will detennine whether it has a bene?cial or
harmful etniogital effect. .

Cm 

l. I. liltill. Survival of ?sh in
tnu?rtltuln. tun-amides. wetting agents and
substatms hit. but Contr. 

.\LDitnls. R. ll'tii. LEA-D residues
acrtal in a Scottish furest. \vmt Res. 

3. . K. (5. ll. Puma". and Al. 
l'thttuci l'Jti?. Mortality among salmon asso-
ciati unit. tilt: fungus Dermoq'stidimn. 1. Fish Res. Board
Can 

t) and a. D. Ftt?st?.
It and I?ftr derivatises in
her l-s-tl (hem. l?JZB-ll??i-l?.

a. \rntun. C.. I. W. HowrtL. A. E. thL. and M. 
I037. Toxicity of 4.3M chemicals to larvae. lain-
act-l fishes. Fish and ??ildlife Sena. 1:5. Dcp. of
Int \tasliiugtun. Spec. Sci. Rep. Fisheries 207.
l'-T

ii twain K. 1967. An evaluation of the persistence of
l) saline in surface waters in the state of Louisiana.
l'tl-c ?su \h-cd Cnnf. 20:343-347.

. i ll?. lit-3". Maximum permissible concentration
nt in bodies of water. Gigiena i Sanit. llyg.

l?-l herbicides.
miscellaneous



in water following

1961. Studies on the fate of
natural surface waters. J.

unit ?at 

8. \xr. C. .\l\TllEP.$ and A. F. ??rst. 10m. Disappear-
ante ot atramte. propazine. and trilluralin front soil and
water. l'toc. 8'0. Weed Calif. 22:367. 

9. 1966. Use of ?As-tagged sodium arsenite in a

study of ideas of a herbicide on ecology. l'toc. of
Sunp. ?13-'29. \?icnna. Austria. pp. 

IO. (I, J. M. Ltmuzxcc. antl H. H. JR.
196'?. The almirption and distribution at C?-labelcd tliquat
in the (Cs-mist": airmms L). Proc. 50. Weed
Couf. 

ll. RentsEraser-r. [962.
obsettatiuus upon estuarine animals exposed to
l?roc. l'..tst. \\'ced Contr. 

Fit'ltl


l?l. R. and L. W. Within. 196']. of
tlnu- teats" testing ditiuat as an aquatic herbicide in
Florida. f?roc. So. \Vccd Cultf. 16:363. 

7-17

AMEN

SQLTEQ

16359

33.

34.

36.

37.

38.

39.

7-18

SCIENCE

l960. Experiments with herbicides in ?shing waters
?'ecd Alt-4r. lS:llT. 71'5.

Baou?x, E. and .\rsnroru. l?thT. Pesticides in selected
western streanrs?.\ contribution to the natioual program.
Pest. Mom. 1. 

BRUAS, F. l93l. The response of certain crops to 
chlorophenoxpcetic acid in irrigation water. Weeds 1:359-
376.

oc joxc. C. 

BLOK, E.

1960. The use of diut0n in

pond pisciculture. \t'eed l8:llS. 733.

G. E.. H. Drum. and E. Hunts. 1964. Tosicire
of aqualin to linger-ling brown trout and bluegills. 
Fish Canto  

P. A. l963. Commercial fisheries imestigatiorrs. 
Fish and Wildlife hert'. Circ. 

BUTLER, P. 1964. Commercial investigations.

US. Fish and Wildlife Sert'. Circ. rates?2:4.
BUTLER, 1957i. Ellcets of herbicides on estuarine fauna.

Proc. 50. Weed Conf. 

Caser. Effects oi sptmittg on plutoplankton in Lake
Rotorua. l'roc. Rotorua Seminar on Water \?L'eetls. spon-
sored by Ext. Serv. L'nit. Auckland. New Zealand. Oct.
15. IQGG. Sl-lO.

R. j. and W. E. Rter-rrt. loco. Control of reeds
and other emergent u'ater weeds itr drainage ditches. ponds
and water courses. The ellects of dalapon on fish. Weed
Abstr. G?Jt'i.

Cons, G. H. H. jrt., j. and
D. E. DANS. l'Jti-l. Persistence of dinuat atrd 
in pools and ponds. Proc. 50. Wted Conf. 171304?32?.

R.. J. l'Or-r. _lrt.. H. Nit-noises. and W.
Barux. The persistence of \il\CX in water and
ht?tlromil. Water Resour?. Res. 

Cor-c. 0. ll. thifi. Sonic responses of ?sh to
herbicides. Proc. So. ?eed (Ionf. 

0. ll. l966. Contamination of the fresh-water eco-
system by pesticides. I. .\ppl. Ecol. 
Cars. 0. 13., j. P. and L. ELLTR. IDtifl. i-Illects
of dichlubenil on pond cruironnrents. Weed ati.

Edits-H55.

U. C. 19155. The cliecb of suits on aquatic
vegetation and plankton in central New York farm ponds.
Down to Earth L?ltl  21:19?20.

B. C. [3:51. The ellects of sodium :Irsenitc and
siltex on the plankton populations in farm ponds. Trans.
Am. Fish. Soc. 94:371?377.

Caosar, I). G. and R. K. 19643. Toxicirv of aquatic
herbicides to anhnin "menu. Science 

DALY, R. H. H. JIL. and j. 
lOtis. Persistence of paraquat. and when
used for etrrasian rmternrilfuil L.)

control. l'roc. Weed Conf. 21:299.
DAVIS. E. A., P. and C. 1963. Eur? of
a watershed treatment with piclnrattr on water qualitv.

U5. Forest Res. .\?ote Rodu Mountain Forest and

Range Experiment oration. Fort Collins. Colorado. -l p.
E. SILVA. S. L. l?til. of 2.4-1) amine
on plankton of tnarshv areas q'prelirninart' nore?r. WeCrl

Alistr. H350. paged.

Eers. R. 1). "Jill. Some preliminary? observations and
?ndings on clients of silrex on the aquatic entironnrent.
l'roc. So. Weed Conf. l9:ll6?ll9. 

Faesrlfl?l. pre-
liminary report on the etlect of some aquatic herbicides
on water qualitv. l?roe. East Weed Comr. Conf. 13:
546?549.

EarnI962. Olisertations on the occurrence and persistence of
2.4-0 and 2.4-diclrlornphennl in lake water. l?roc. No. East
Weed Contr. Conf. 

Earn. 5. I). am] 0. 

1963. Some effects of 2.4-0

and 2.4-DCP on drinLing water l'roc. East
Weed Cunt. l7. Min?470.

F. F.. E. K. ?moon. antl R. 1963.
Preliminary- ?eld trials of acrolein in the Sudan. World

llull. i'iz?llil?L'IS. 
it retiew of duratine 
Prue. East Weed . Corrf. 

Health Org.
H.
weed Control.

505.

40.





~13.

H.

46.
47.

4d.

49.

67.

Ftuut. I966. Persistence and distribution of nronumn
and ill in aquatic elurtonment. eeds Hill!)?
our)

huts. P. R. ll. lloocsox'. R. D. Cnuts, 1967. Residues

of two herbicides in \ialer irrigation canals. Weeds
15:353?355.
A. and R. D. COHFS. [967. Herbicidal residues

Weeds 
l'Jtia. \'r'eed Control Hand-

in pond water and lndrosml.

Faun. and Evans.
book. Vol. I. Principles.
Oxford and Edinburgh. 45? 

C.nt.ort. J. and Hour]. 1962. Three \?ears' results with
for aquatic plant control in Oklahoma. Down to
Earth 

A.
tions of sodium at'senite on and the aquatic en-
t'ironment. Team. Fish. Soc. SIESELHOO. 

l'JtiT. Ellects ot' diquat on and
their food organisms. Proe. l-islr Cult. 29:07?74.

ll. lfltii. l'roiett 2i: L'nrlesirable u'oudt- plants.
Res. l'r'og. Rep. est. Weed Contr. Cont. 

Gunner. O. l. l?Jo'T. Cortlau?litlaliun of a sin-fact: water
supplt source of the Slrortantlt' Station with herbicide Est-D
and ester. Gig. Sanit. 32:97-93.

(2:21.305. A. R. l963. Public health aspec'ts of weed corr-
trol in potable water supplies. l'roc. So. \'r'eed Conf. l6:
420?424.

GuamCox.
The persistence of four herbicides in pond renter.
So. Weed Conf. 

Ct'sc. L. R. lfllil. Aquatic rec-ed control research in lndiana.
Res. Rep. No. Cent. Wtul Court. Conf. 

R. X. 1969. Testing' dittrorr as an economical
herbicide itr recreation lakes. Proc. 50. Weed Cent. 22:
321?3123.

lhrtor'. j. L. lilliti.


Proc.

Effect of 'l'nnlon herbicides on aquatic
chain organisms. Down to Earth 

Hue. C. L. and R. 9. Curt-mu" l9t5l. Ellecls of the herbi-
cide siltex on benthos farm pond.  Wil-llile .\lartage.
25:333-317. .

ITGG. [affects of di-?iuat on antic-.23: ins-tits
and related animals. Econ. Errtunrol. 5921320413.

Hou. W. 1963. Chemical weed control irt ditches: trials on
the control of plants. Weed .?\bstr. l2:lU0. No.
50?. paged.

HorsE. W. 13.. H. Goonsox. ll. aml K. W.
Doom?. Assessment of et'nlugical effects at cut-mite
or repeated use of herbicrdes: littal report. .\lidu'est Res.
lmt.. Kansas (in. 3:39 p.

and Garcon. lfltil. Results of 
and two of srl\e\ tor the control ut aquatic
plants in Oklahoma. l?roc. 50. Word Conf. 

HOMER. A. 1963. in \teteltt of lollmving
aquatic ?aeration control using ttre herbicide silt'ex. l?roc.
()Lla. Acad. Sci. 82232-257.

J. S. The toxicitv of ?sh of tlill'er'ent forum-
lations of Lilli-'1'. and silvex. Pine. 50. Weed
Cont. 53163?2156.

Ht'cnrs. j. S. and j.
to bluegill 
53.

H. (Chnr.).
Soc. of Amer. [?93 

New: E. G. 1961'?. llioloeical
Scienti?c Aspects of Pest Control 231?262.
\t'ashittgton. 1112. 470 p.

T. nuts. 
of phenoxy herbicides.

Variations irr to\.icits'
ll: 
1967. Herbicide Handbook. Weed
Magni?cation of Pesticides
.\cad. Sci.

5. and T. Elfects of pesticides
and herbicides upon microorganisms in soil and water
under waterlogged conditions. Bull. Scr.

tjapan). Ser. ll. hid-90.

)scortosst. F. The to\iculn_eical problems of controlling
weeds in canals. Weed l-li'JtiT-tltil'l. 1306. 
Jonxsox. R. E.. T. C. Carmen. arid E. ll. l961.

Indicator species near top or food chain clro~en [or assess-
ment of pestitide base [ads in lish and 
Outer. and in estuarine enrirornnent. l'est. Moon.
I. l:T?l3.
Ktx?mi. E. E.
to and birds.

Volume l8. Issue 6 (November). 1970

1828

I939. Tordon herhicides?Er'aluation of safety
Dorm to Earth 

Blackwell Scienti?c l'ublications.<

Some elfecrs of sublethal concentra- 

L.




0

68.
69.

80.




a2.

83.
8-Law-amen. 

MULLISOX

Kuamttx. G. C. '53. \l'eed Control: a Science. john
?'iley  Sons. Inc.. New York. ~12! 

Kocrixn. Y. 1967. Experimental data for the 
standardization of the dalapon in bodies of water.
Gig. banit. 32:7-11.

5. anti D. B. Wreuzrs. 196-1. Ell?ects of 2.4-1)
and 2.4..5-T on water qualitv after spraying treatment.

US. Forest Sen. Re~. Note 32:1?i.

1937. Recent on the we of
sodium arsenite as an aleacide and lb eilects on ?sh pro-
duction in ponds. Southeast. Assoc. Carrie and Fish Curit-
missioners 11:231-956.

Lass-user. j. 51.. R. D. Bucxat?nx. and G. Brtsen'. 1961.
Aquatic herbicide eialuation tests using plastic pools.
Proc. 50. Weed Conf. 14:309?316.

1962. Aquatic herbicide data. Agr. Hand-

book 231. Agr. Res. Sena. Dep. of Agr. 471 p.

1963. Graphic presentation of aquatic
herbicide tlata. 1?roc. ho. Weed Cont. 15:566?373.

thnsacnav, H. L. 1961. Considerations regarding the use
of aquathol in potable watersheds. 1?roc. No. East Weed
Contr. Conf. 15381?184.

thocx. G. and A. Mt?LtIn. 1963.
the efft-Ct of diesel Oil on the soil.
791-793.

K. 1960. Some limnoloeical investigations
on the long-term use of sodium arscnite as an aquatic
herbicide. 1'roc. Can". Weed Contr. Conf. 17:30?31.

R. 1961. Obsenations on the Use of
2.4-0 granules in ?sh hatchery ponds. Res. Rep. Centr.
Weed Contr. Cont'. 18:11:13.

Martin. L. A. and R. COLMEI. 1959.
acid by soil bacteria.
biol. 52251-260.

Matoxcr. T. E. 1958. Control of Ale-ac with chloronhcn?
dimetht'l ttrea. J. .lmer. Water Works Assoc. 50:417 2'2.
Martsrox. R. 3.. D. W. Seneu?s. T. Museums. and L. 
lira-i. Amitroie concentrations in-crerk watt-rs
from an aeriallt' sprated watershed sub-basin.

Pest. Monit. j. 

MAth 1. and l. (intenutsun. i967. Toxic chem-
ical low! in water and and treated with itemi-
citles. Gig. Sanit. 32:96?93.

L. 1. 1960. Aquatic weed control.
Zealand Weed Contr. Conf. 13:58?61.

Moexr. D. I. and C. E. SrEt-uex. l9t57. A method for estab-
lishing acceptable toxicant limits for fish-malathion and

Further investigations on
Allg. 13:

Decomposition of
Can.  .\licro-

1?roc. New

't'he butoxtethanol ester of 2.4-0. Trans. Amer. Fish Soc.

96:185?193.

Aft'Lusox. W. R. 1966. Some toxicological aspects of silvex.
1?roc. So. Weed Conf. 19:4215?l33.

NATIONAL Acaorxtr or mner RES. 
1966. Principles of Plant and Animal l'e-t Control. 2.
?Feed Control. .\ratl. Sci._ \Yashiugtou. 11C. 47] p.

j. 1965. Methods of wildlife hazards.
Proc. Brit. invest. and Fungic. Conf. 5:342?537.

max, 1. F. anti  War. 1966. some ecological obser-
vations iu the use of paraquat and diquat as aquatic
herbicide. 1?roc. Brit. Weed Contr. Cont. 532-535.

NEWMAN, J. F. 1963. The ecological ctfeets of paraquat and
diquat when used to control aquatic weed. Weed .-\bstr.
17:361. No. 2211.

H. P. and j. R. THOMAS. I963. Pesticide per-
sistence in public water. their anti rentmal. pp.
181?189. In Research in Pesticides. Academic Press. 1nc..
New York.

NtsutL'cut, Y. and Y. 19137.
cide ingredients to some fresh water ortzanisms.
Insect Contr. 32:5-11.

NOLL. D. C. and R. 1961. interim report on
aquatic need control of recreational waters abme potable
water. Proc. East Weed (Inntr. Conf. 

Nouns. L. Nut-rox. and I. 1967. Stream
contamination with amitroh: from fore-t spray operations.
Res. l?l'og. Ken- West. Contr. Ctmf. pp. 33-3".

NoRaIs. 1.. 19-1". Stream contammatton hv herbicuh-s
after fall rains in fortxt Lind. Res. Pin-.1. Rep.. ?Cat. but".
Of ?'t?ed lici. pp. 33-0?.

Oxlrro. K. and T. Oxtao.

Toxicitv of pesti-
Bull. Inst.

396'). Intiuence of diluted sea

Volume 18, Issue 6 (November). 1970

100.

101.

103.

i07.

103.

109.

110.

111.

119.

. 1. C., W. F.. hl't'l?El. and R. R. joussox.

. Sum. M. Ktru'un. and T. Kirsrrax 

EFFECTS AQUATIC ORGANISMS

water on the phuiolotzical actititv of hairs-neck clam.
l'cncrupir l?f?O'H?t. and the tou'c effect of an herbicide
1?Cl?, pentachiote-pheuate. Bull. Tokai. Reg. Fish Res.
Lab. 44:31?10.

Patina. S. j. and ii. Wonru. 196.3. The effecrs of tri-
tluralin on hsh. 1'roc. (so. Weed Cnnf. 15:469?473.

Penn's. C. F. 19:53. Elleet of herbicides on heterotrophic
microorganisms of ponds. Wet-d 11:330. 13-37.

E. 1935. The effect of the wee-dicith Huron upon
the ?ora and fauna of two experimental areas of Long
Pond. (.nunu. New York. 1?roc. East Weed
Contr. Conf. 12:335?343.

PIERCE. M. E. 1939. Further studv of the effeu of the
weedicitle Kuron upon the ?ora and fauna of Long Pond.
Countv, New Yark. Proc. East Weed Contr.
Conf. 13:310-314.

Menu. E. 19-60. Progress report of the effect of Enron
upon the biota of Long Pond. Countv. New York.
Proc. No. East Woed Contr. Coni. 14:472?473. 

PIIRCE. E. 196-0- :1 study: of the effect of the weetlkiller.
2.4-D Granular. on three experimental plots of Long Pond.
Countv. New York. Proc. East Weed Contr.
Conf. 14:483-457.

PIERCE, E. 1961. studv of the ctfect of the weedkiller
2.4-0 Aqua Granular on six experimental plots of Long
Pond, Countv. New ?t'orL. 1?roc. East Weed
Contr. Conf. 15:539?344.

PIERCE. E. 1962. A comparatite study of the application
of three weedicides. Kurosal G. Kurosal 5L and 2.4-D ester
to three areas in Long Pond. County. Proc. 
East Weed Contr. Cont'. 15:135?141.

PIERCE, E. [963. Progress report of the application of
fenac granular to six small plots in Long Pond. 
County. New York. 1'roc. No. EaSt Weed Contr. Conf.
17:431?133.

l?lERCl.. E.
ponds on Vassar Campus.
Conf. 20:470?475. 
PIIRCF, E. 1967. Study of a second sear application of
fenac to two small ponds on \?assar Campus. 1?roc. East
?'eed Contr. Coni. 31:330-533.

E. time. The efiect of several herbicides on
eight test areas in .\'obska Pond. Woods Hole. Massachu-
setts. 1'roc. East Weed Contr. Coui. 22:193-203.

Picker. E. 1969. The effect of a combined formulation
of 2,-l-D and endOthall upon a small pond. 1'roc. East
Weed Contr. Cont. 23:376-379.

PIERCE, P. C-. j. E. Farr. and H. Yawx. 1963. Field
evaluations of newer aquatic herbicides. 1?roc. So. Weed
Cunf. 18:497-556.

P. C. MW. Results of ?eld testing newer aquatic
herbicides with emphasis on spot treatment applications.
1'roc. So. Weed Couf. 19:374-373.

Pnowsr. A. 1960. Notes on the toxicity to ?sh of some
common agricultural insecticides and weedicides. Weed
Abstr. 16:33. .?sot paged.

Rao, K. N. and M. 5. Meters". 1963.
repmru Fair with herbicides in Andhra Pradesh.
Abstr. 17:36-1. 2233.

Rawts. C. K. 1965. Field
certain estuarine animals.

1966. Application of fenac to four small
1'roc. Xo. East Weed Contr.

Con trol of [homoca
?'ch

tests of herbicide to\icit\' to
Chesapeake Sci. 6:150?161.
1963. Will
the use of 233-1? to control strearnsitle seizetation cott-
taminate public water supplies? I. Forest. 56:914-913.
Rnxuu?r. K. C. l?JtiJ. Herbicitlal treatment of watersheds
to increase water yield. Proc. East Wad Contr. Coni.
19:546?551.
D. X. 1964.
treated with diquat.

R.. E. A. Cut. and H. H. jR.
l?Jti7. Effect of paraquat on growth of Srinau?uiu 
in liquid culture and soil. l?hstopathnlugv 57:911-915.
lT-tii? Studies
on the pretentron of poisonin in agricultural cbt-iuicalscl?l.
lullut nee of herbicide isodiuut pentathlorophenni- ~preat  on
paddstit-ltl regions with riser water. Rep. Hokkaido lust.
l'til). llt?alth 
acnuxntz. ll. (3.. jtt. 1967.

Observations on some acid water lakes
Proc. East Weed Contr. Couf.

Microbial degradation of pesti-

749

18

MOO

128.

129.

130.

131.

?1

cides in aqueous solution.
1701-14.

SUIES,  C. 1961. Control of the Polar-"gnarl
crisp?: in bath 110m": and static situations with cudothall.
l'roc. East Weed Couf. 15:356-559.

Ssn?rn, C. E. and B. C. Isou. [1167. Investigation of effects
of large-scale nl?pllc?lulti of 2.1-1) upon aquatic fauna and
water qualitt'. l'est. Monit. 1:16?21.

1. Water Poll. Contr. Fed. 39:

. SNOW. R. 1963. preliminary report on the control of
l?itltophora with simatine. Proc. 50. Weed Conf. 15:329-
333.

Observations on the use of disndium
Res. Rep. No. Centr.

E. W. 1961.
entlothall in hatchery ponds.
?'eed Contr. Conf. 15210?1.

E. W. Observations on the use of dismliurn
endothall in ?sh hatcher} ponds. Weed Abstr. 12:197. 
1036.

E. W. and 0. L. 3111115131 1931.
tions of arsenic for certain aquatic organisms.
Amer. Fisheries Soc. 61:225-239.

Tumxr. R. F. and L. NORRIS. 1967.
cities and diesel oil carriers in forest water.
Herb. in Slanazc. pp. 81-88.

W. M. and R. D. 1965. Preliininarv
study of the effects of diquat on the natural bottom fauna
and plankton in two subtropical ponds. Weed Abstr. 14:26.

Lethal concentra-
'I'r.tns. ot

Residues of herbi-
Proc. Syrup.

1-1-1.
TAYLOR. L. 1966. Catoron a new aquatic herbicide.
Hyacinth Contr. 1. 5220?21.

TAYLOR. L. 190i. Dichlobenil aquatic herbicide for use
in non-?owing WJICT. Proc. 50. Weed Conf. 19:376?377.
THIEGS. B. 1935. The stability of dalapor. in soils. Down

to Earth ?(Eel?t.
THOMAS1963. 

ester of 2.4-D in the control of Eelgrass (.Z'osrem mcrtam L.)
and its effects on outers (Crauostrm virzt?mm Cruelin)
and other benthos. Proc. East Weed Contr. Conf.
22:186-193.

L. 1962. 'Present and future of aquatic weed
control. 1?roc. Last Weed (.uutr. Couf. 16:1?8.

F. H. l-Ju'J. Ueluliattott tn \tetnani. The eco-
logical conscquences of the defoliation program in \ietnant
are assessed. Science 16327794813.

U.S. Der. or her. Report of the Committee on Water
Quality Criteria. Fed. ?'ater Poll. Contr. Admin. \1'ash-

-ington, DO. 234 p.

i 5.

135.

136.

137.
133.

139.

HO.

1-11.
1-12.

1-13.

1-1-1.

1-16.

1-17.

SCIENCE.

U.S. Den. or 1\l.
Wildlife hilltiiL?S.

1053. Fish and Wildlife Sert ice. Pesticide-

Rcticw of Full and Wildlife Service in-
\estit_.vation tluttn: 1961 and 10:52. Circ. 167. 109 p.

Du: or lxr. 11"31. Fish and Wildlife Nt'lWi?fJ. Pesticide-
Wildlife Studies. Reiiew of l-i~h and Wildlife Service in-
vestigations during calendar t'cat? Circ. 190. 130 on

U.S. Dun or in". 1955. Fish and Wildlife Service. Pesticide- 
Wildlife Studies. (inc. 1326. 77 p.

L'snie. G. 0., M. Ftaooq, K. Dmoon. L. and 13.. 

D320. 1963. Field trials in limpt with acrolein herbicide-?i
molluscicitlc. Health Org. 1111?. seem?:60. ?1

\?Ath. C. C. 19643. Persistence of 2.13-t1ichlorolienzonitri1e
in Aquatic limironments in Oraanie Pesticides in the Ean
\?ironment. A Smt. Amer. Chem. Soc. in Chem-C?-
istn,? Series 60:271?270.

?Tamra, C. R. 1961. Toxicoloeical effects of several herbi-i?
cides to bottom dwelling ?sh-food organisms in Missouri.\
ponds. Res. Rep. Centr. Weed Contr. Conf. 

C. R. 196:1. Endothall derivathes an aquatic herbi-
cide in ?shert' habitats. Weeds (I

Wiutrtt, C. R. 196-1. Diclilolienil as a herbicide
habitats. \\'eeds 122267-2139.

WALKER. C. R. 195-1. Simazine and other s-triazine com-
pounds as aquatic herbicides in ?sh habitats. WeCds 12:
134-139.

?Hutu. C. R. 1965. Diuron. fcnuron. monuron. neburon,
anti TCA mixtures as aquatic herbicides in ?sh habitats.
Weeds 13:297-301.

Wcmox, L. W. antl R. D. 1967.
floating aquatic needs with ametryne.
Conf. 20:312?318.

tt'ittnoax. T. L.. 
and herbicidal compounds to striped bass.
31:27-32.

?tasttaw. R. C. COLDIIERG. and D. 1967.
The determination of the ioniration constant of 2.1-1) in
water. Water Resour. Res. 3:311-316.

th.r.. l. 196:1. plant conttol with femur. and
sitnazine in Ontario farm ponds. Proc. East 
Cuntt'. Conf. 21373-133.

.1. in. anti 31. Run. 31'th happens
to ltet?ltieitles when applied to a watershed area?
Proc. No. East Weed Contr. Conf. 15:396?101.

?t'ro. R. R. 19117. Dissipation of diquat and paraquat. and
effects on aquatic weeds and fish. Weeds 15:12?16.

in ?sh

The control of
Proc. 50. Weed

1969. The toxicitv of nine therapeutic
l?rog. Fish-Cult.

1970

8282

Volume 18, Issue 6 (November).-

PROGRESS REPORT ON THE use or meow, 2,h-D and 2,11, 5 TP Gamma
- AS AQUATIC 

by 
Roy R.'Younger 1
Assistant Fisheries Biologist
N. J. Division of Fish and Game

The purpose of this report is to summarize the results
of three year?s experimentation using Kuron, and one year's
progress in using 2,h-D and 2,h-5 TP granules as aquatic herbi-
cides. - 

Kuron

 

The first experimental work with Kuron was conducted
at Ramapo Lake, Passaic County in 1956. g/ The following year
thirteen lakes and ponds were treated. The results of these
experiments have been presented in a previous paper. 2/

At Ramapo Lake, the herbicide was applied at a con-
centration of 2.5 p.p.m. to control water milfoil, 
fanwort, Gabomba caroliniana; and white water
lily, Mumphaea odorata. No further treatment was necessary in
1957, however, in 1958 the treated areas were completely rein-
fasted with the same noxious weeds.

 

 

Good control of the most predominant species - water
milfoil, sp., yellow water lily, Nuphar sp., and
white water lily, sp., was obtained in five ponds re-
ceiving total applications of Huron in 1957 at concentrations
ranging from 0.5 to 2.5 p.p.m. In 1958 these ponds needed no
further treatment because the water milfoil had not reappeared
and the water lily stands were greatly reduced. Considering
the size and extent of the lily root systems it is not diffi-
cult to understand why total control in one application is not
possible.' 

Plant reinfestation seems most likely to occur, at
least in New Jersey, from seeds and plant fragments entering
?from untreated lakes upstream, rather than from the-germination
of seeds after the application of Kuron. There is some evidence

 

ly?iThe author is now employed as a technical representative of
Halco Chemical Company, Kenilworth, H. J.

g/ Huckins, Robert K. 1956. Job completion report, Project
Aquatic weed Control, unpublished. 

2/ Younger: ROY R. 1957. Preliminary studies using Huron as an
aquatic Weed Control Conference, New York, January, 1958;?,
All this work has been conducted under federal aid in Fish?&

Wildlife Restoration Acts, New Jersey project numbers F-1oan

   
  
   

   

.. 

   

   

. . lib-<14:
11?   
"21rd!" 7141?- ?i 
I 155-3349.. - 

  

. . . .. rim-'1 rat-7i 
l, . 
-.. I a, 



18988171 

324.

reduced prior to treatment greater plant destruction was
achieved. Therefore, it is recommended that lakes and ponds_
receive total treatment and that the lake volume be reduced;
to insure sufficient contact time. .

The concentration recommended to obtain adequate
control with Kuron is 2.0 to 2.5 p.p.m. The Dow Chemical? 1
Toxicological Laboratory has determined the L.D. 50 for the
emerald Shiner, Notropis atherineides, is 7.0 p.p.m. The
recommended concentration is far below the toxic level and
fishkills should not occur, providing other chemicals are 
not used before or after treatment.? Furthermore, damage to
shoreline vegetation should be negligible at this concentra-
tion. 5/ 

 

@3993le M00

.
i

3. Bottom application

Spraying the exposed bottom with Huron at the rate
of gallons per acre has proven successful in one instance;
It might be to more fully explore this technique, 
thereby reducing the cost and making it possible to spray at 
an earlier period in the spring. 

Conclusions

The results of three years of experimentation would
seem to indicate that longer periods of control are obtained
with Kuron than with other herbicides. 

2,h-D and 2,h,5 TP granules

As a result of Dr. Grigsby's work in Michigan,
the New Jersey Division of Fish and Game became interested
in further field testing of 2,h-D and 2,h,S granules.
Thirty-six plots containing 2,500 square feet (.057 acres)
were laid out to evaluate the effectiveness of 2,h-D and
,2,h,5 TP granules of either 15/30 or 8/15 mesh size. The
granules were tested in three plots and the in
thirty?three plots.

 

5/7 There have been some fishkills in the past. In one
instance the fishkill was caused by the addition of
copper sulfate for algae control prior to treatment with
Kuron. Another fishkill in a farm pond was attributed to_
the rinsing of spray equipment previously used to apply 
Halathion;~ A third fishkill-has never been explained 
-due tc.so many extraneous factors which occurred either_* 
prior to or immediately following application. Ti He

Grigsby, B. H. and R. Hamilton: 1958. New techniques?

- in the application of herbicides for control ofhaquatio-Vl:r

'23ja?

 
  
 
 
 
  




plants. ,Presented_at,the_1958 meeting of the'Weed 
103: zJanuar?iwl9S??
 :3  i 

   



    

 

Conclusions:

. The advantage of using granules is ease of dis-
tribution since no cumbersome Spray equipment is required.
This makes it possible for lake front property owners to
treat their portion of the lake.

-=If the mechanics of the grannles can be improved,
and if the rates of application can be determined, they 
will definitely have a place in aquatic weed control.

J3.  
 

    

"#391? 


. 

 

939.8315! 

    
        

Table I Contd.

328.

WW I 42 85.2.7. 

16285

smear OF 2,114) and 2,115 TP my mums DI 1m: Jaws Hm 1958

a

 

 

Area treated Type of vegetation Control
Applegate's Pond Fannort, Cabonba?sp. None
Honmouth County Spatterdock, ?gynar Sp.

Uater milfoil,

Medi'ord Lakes Myriop?mjllum Sp. ?one

Camden County

Musconetcong Lake
Morris County

unite eater lily
Manaraa sa-
2,b-D control only

- 8
Hater milfoil, Riki?? 

 2,h,5 TP control with
. 9 lbs/acre l/

 

pratcong .

Hater milfoil,

619- Good control at

 

 

 

Spattardock,
Susser County Nu har Sp. all rates 2/
Pond weed Potmegeton 5p;
Grovers Mills Fanmort, .
Mercer County? Cabomba sp. lone 2/
Este rule Lake Bladderwor 
??f}antic Utricularia sp. 
Saxtons Falls Hater milveil .
warren County ?cue-l/

sp . I

 

 

 


9'



At time of application no Vallisneria was observed. After treatment and destruction
of the Vallis?gria took over the treated area. 
Regardless of rates of application, good control was had for a short period of time
Since weed fragments from untreated-areas settled to the bottom, putting out advan?
titious roots, thus reinfesting the treated area. 

In these plots the 2,h-D was sprayed on soft attaclay which broke down immediately
upon contact with water.

ed Control Conference

Reprinted from the Proceedings of the Thirteenth Annual Meeting
of the Northeaster

.
a,
.

Ea
January 7, and 9, 1959

 

 

IQ)
CL)
..


 

ACD File No. HA-8

REPRIHT OF PAPER PRESENTED AT THE

THEIPTH ANNUAL MEETING
OF THE

HORTHEASTERH WEED CONTROL 

Hotel New Yorker

New York City

January 8, 9, 10, 1958

{79.6883 moo

?62867

 

by -
Roy R. Younger
Assistant Fisheries Biologist
New Jersey Division of Fish and Game

In the summer of 1956 the New Jersey Division of Fish and Game,
under Dingell-Johnson Project No. Aquatic Weed.Control, experi-
mented with Kuron on one-acre plots in two northern New Jersey lakes.
The chemical called Kuron - Prepionic Acid - PO
is a hormone-type growth regulating compound which is absorbed by the CD
leaf and translocated to other parts of the plant. The above mentioned?'0
plots were heavily infested with water milfoil, hetero- 2%
fanwort, Cabomba caroliniana, and white water lily, L:
are a. One week after application good control of the submerged 
species was evident' the emergents were not materially affected.
(Huckins, 1956, ms.5 It was felt that no further treatment was necesq
sary after examination of the plots in the spring of 1957.



The results did seem to indicate that further evaluation was 2
desirable and a testing program was initiated in the summer of l957_/.

Hethods and procedures

Lakes and ponds, in order to be included as part of this testing
program, had to meet certain requirements:

1. As a group they had to reprpsent a cross-section of existing

'lakes and ponds throughout the state with regard to weed species, both

emergent and submerged, alkalinity and hydrogen-ion concentration.

2. They had to have a woody shoreline so that the effect of
Kuron on woody plants could be evaluated.

In treating a pond the procedure was to calculate the active
ingredient in Kuron required to produce the predetermined concentration
in parts per million.

To determine the effectiveness of Kuron at various concentrations
the herbicide was tested at 0.5 1.0 2.0 and
2.5 p.p.m. One reclaimed lake in southern New Jersey was treated at

305 ngcm.

The herbicide was applied by a Bean Estate Sprayer, skid mounted,
with split intake lines and the by-pass was used for circulation and
agitation. A spray boom was devised and a thirty-gallon drum served as_
a spray tank. The entire rig was mounted on a 11.5 feet aluminum boat
and was operated by two men. The herbicide was applied in rows with a
slight overlap between them. 

 

(- 3/ Misc. Report's21, New Jersey Fisheries Laboratory, Division of

Fish and Game, Milltown, New Jersey.

g/ This program would not have been possible without the assistance
of Dr. E. Evaul and Neal Munch of the Soil Conservation Service
who obtained ponds for testing; Dr. Mark Wiltse of the Dow
Chemical Company supplied the Kuron.

18288_

 

333.

After the ponds had been treated water samples were taken and
forwarded to the.Dow Chemical Company for Karon analyses. The water
samples were taken from two to twenty-four days after application.

Table I .

Ponds were checked at seven day intervals for the first month
with checks thereafter.

Findings

During the course of the summer eight lakes received total appli-
cations and five lakes received plot treatments of a half-acre in area.
A summary of these applications is presented in Table II.

The concentration of 0.5 p.p.m. was effective on white water lily,
The submerged species were only affected. At 1
p.p.m. good control of water weed, Anacharis sp., yellow water lily,
Nuphar, and mud plaintain, Heteranthea sp., was obtained. Complete
control of water milfoil, and bladderwort, Utricularia,
was achieved with concentrations of-2.0 p.p.m. and 2.5 p.p.m. rom
these data there is every indication that adequate control of most
species can be obtained at 2 p.p.m. however, further testing of the
effects of Kuron on pondweed, Potamogeton, is necessary since none of
the concentrations employed controlled this species. Duckweed, Lemma
minor, turned brown forty-eight hours after application of Kuron and
then recovered fully.

 




As to the effects of Kuron on trees, either in the water or
iurrounding the lake, no damage was evident on such species as silver
maple, black gum, pitch pine, willow.nnd white cedar.

Since this herbicide contains phenol the possibility of its
tainting fish flesh was considered. Fish were collected from a treated
area and taste tests revealed that Kuron did not taint in any manner.

As mentioned earlier, measurements of pH, alkalinity and tempera-
ture were recorded. None of these water characteristics appeared to
,affect the herbicidal effects of Kuron.

Observations suggest that Kuron is slow to act. Seven days after
treatment submerged plants appeared wilted and emergents were curled.
Fourteen days after treatment open areas were visible; submerged vege-
tation had started to rot, while the emergents continued to curl. It
was not until twenty-one days after treatment that the submergent weeds
were completely disintegrated and leaf and stem systems of the emergent
vegetation had broken free from the-roots. One month after application
root systems of both types of weeds were observed in windrows along the
shore.

Since the affected plants did seem to break down quite leWIy,
there was some suggestion that no serious oxygen lag occurred. However,
"this cannot be substantiated since an insufficient number of D. O.
"leterminations was taken. Observations also suggested the absence of
Algae blooms, sometimes characteristic of certain herbicidal applica-
"On3, caused by the release of essential nutrients from the rapid

eakdown of plant tissues. However, this lack of algae blooms could
also indicate possible algacidal effects of Kuron.

m2 3 3

 

Further evidence of the slow breakdown of Kuron was indicated by
the water samples which were forwarded to the Dow Chemical Company for
Kuron analyses. In the total applications the concentration of Kuron
remained fairly constant while in plot applications the concentrations
were affected by dilution, or a build~up in stream application. Resi-
dual effects cannot be determined until next spring when the viable
seeds, if any, will germinate.

Plot applications were not as effective as total applications
since reinfestation was possible when shoots and seeds from untreated
areas floated into treated areas. The possibility also exists that
once the vegetation has been destroyed other aquatic species, either
desirable or undesirable, may over?run the treated area. Total applica-
tions were most effective since reinfestation was less likely to occur.

Summary and conclusions

The experiments conducted to date have more or less been designed
to determine the herbicidal effects of Kuron on aquatic vegetation.
Success has been obtained on the most predominant wood species in the
thirteen lakes and ponds so treated, using a concentration of 2.0 p.p.m.
The period necessary for total plant destruction is normally twenty-one
days. The usual algal blooms were not observed following tneatment
with Kuron; preliminary data suggest no serious oxygen lag. This could
be attributed to the slow breakdown of the plant tissues but the apparent
scarcity of phytoplankton also suggests the possibility of algacidal
properties.

None of the concentrations employed have affected the trees either
in the water or along the shoreline. Neither pondweed, Potamo eton,
nor duckweed, Lemna minor, has been effectively controlled by any con-
centrations of Kuron. .

There are many important questions still to be answered regarding
the possible use of Kuron as an aquatic herbicide. These questions
include: At what time of year will application achieve the most
effective control of weeds? Does this herbicide have any undesirable
side effects on certain micro fauna and flora important to game and
pan fish species? Will it control filamentous algae? Does it have a
residual effect? Will its cost affect its use?

The results to date have been promising and the proposed questions
are in the process of being investigated. In the meantime Kuron can be
considered as.one of the most important aquatic herbicides on the
market since the development of sodium arsenite.

lifts  

$98888



12.73331

Table I.

AMOUNT OF KURON DETECTED FROM FIVE TREATED
PONDS 2 TO 2? DAYS AFTER APPLICATION

(Analyses conducted by de Chemical Company)

 

Kuron applied

No. days sample taken Kuron detected

 

Name of Fund p.g.m. after application . p.g.m.

Rte. No. 1 2.0 2 days 2.0
Golomb's Pond 2.5 2? days 1.5
Keswick Pond 2.0 14 days 1.8

New Waywayanda 2.0 3 days 2.1
Panther Pond 2.0 8 days 'Less than 0.5

 

896883



'Y?a?iC-llg

L?s

 

 

 

 

 

 

 

 

 

Table 11 SUMMARY OF KURON APPLICATIONS - 1957
Name of lake Kurcn applied Tot.a1E. ?Dominant weed 
or pond p.g.m. QH p.p.m. Temp. species Degree cf Control
Taylor Pond 1.0 2.2 13.0 86? Heteranthera dubia Complete
Anacharis canadensis -
Potamogeton crispus Excellent control of sub-
Mohawk Lake 1.0 8.h 82.0 78? Typha latifolia mergents, no control of
Scirpus americanus emergents except those
Sparganium sp. individual plants which
Lemna minor came in contact with Spray
Nuphar variegatum
Anacharis canadensis Excellent control of both
Bell Lake 1.5 8.3 105.0 68c nghar variegatum species
Eriocaulon septan-
Ludlum Lake 2.5 6.1. 14.0 81? gulare
(plot) Excellent control
-Reinfestation
Vallisnerid americana
Nymp?aea odorata
Cape May 0 PotamOgeton sp.
Park Pond 3.0 6.6 12.0 79 Anacharis canadensis Excellent control
Utricularia
purpurea
In doubt. Pond was previous-
Barker Pond 0.5 7.3 6.0 78? 1y treated with sodium
arsenite
odorata
Potamogeton sp.
Vallisneria americana Only Species affected was
Applegate Pond 0.5 7.0 27.0 80? Cabomba caroliniana water lily



 






~32






 

896883 

 

 



H3

CAD

 

 

 

 

 

 

 

 


Table II Cont.
Myriop?yllum
Galomb Pond 2.0 6.7 12.0 81? 
. . Nuphar variegatum Excellent control
ggirpds Sp. - 
Nuphar variegatum 
Cabomba caroliniana Plot #1 only Nuphar. Plot
Foo Lake 0.5?p10t #1 5.L 53.0 80? and Plot #3 good control
. Q.1_plot #2 Lcmna minor No control of Lemna
3,0-p10t $3 Anacharis canadensis
Utricularia purpurea .
Keswick Col? odorata_ Excellent control
onv Lake 2.57 5.6 2.0 78? Pocamogeton 59. except for Pctamozeton
-
Panther Lake 2.5 8.8 103.0 74? Excellent control
odorata
New Wayuay- Excellent cortrol
anda Lake 2.0. 8.8 107.0 76? 
Scirpus I
MacDowell americanus Only Scirpus was
Pond' 2.0 6.8 76.0 80? Potamogeton Sp. controlled


1202

SUMMARY OF TOXICOLOGICAL AND ANALYTICAL INFORMATION

ON THE CHLORACNEGEN IMPURITY FOUND IN SOME SAMPLES OF 

OCCURRENCE AND IDENTITY

HIM

Several impurities can occur in arising from their



presence in The; can be minimized

by proper operating procedures. The most important impurity

-4



which causes the chloracne problem is:

.

symmetrical tetrachlorodibenzo-p?dioxin

 

 

TOXICOLOGY
Note. All doses given as MICROGRAMS (10-6 grams)

 

Skin Contact

Human

1. Known to cause severe chloracne by skin contact.

Very small amounts can cause the lesions.

182174

 

 

 

Rabbit
. . . If.
1, Inner surface of rabbit ear treated various 
 
levels of the chloracnegen in 100 mieroliters (ul) -a
of a test solution in acetone benzene or chloroform f?
H?h
on an area animals per doselotal Dose Applications CD

0.2 ug repeated no effect
0-5 U3 repeated marginal effoct*
1?2 ug (sometimes from1 usually causes an effect
4?8 ug lone applicatio?E= almost always causes an

effect, usually severe,
but not lethal

2. Shaved belly of rabbits treated with 10% solution
in acetone, 5 animals per dose. L050 by sgin
absorption 275 ug/kg body wt.
Mortality generally occurred 12?22 days after
dosing.




palathlon, 2, the D5
by skin absorption has been reported to be:

0

rabbits 40,000 ug/kg body wt.
rats(M) 21,000 ug/kg
ratSCF) 6,800 ug/kg?

Intraperitoneal

When administered as a 10 mg suspension in corn oil to

rabbits (5 animals per dose level):

I.P. LD50 283 ug/kg body wt.

 

*Follicular prominence and/or epithelial thickening begins
1 to 3 weeks after exposure.

confidence limits.

16275

 

{for parathion, the intraperitoneal LD50 has been

(1)

reported to be:

E,

I

b_

5,500 ug/kg ad
guinea pig 12,000 ug/kg
12,000?20,000 ug/EE

ll

rat

5



dog

Acute Oral (single dose)
Rabbit

When administered as a 10 mg suspension in corn oil

(5 animals per dose level): Acute oral LD50 100
ug/kg. Two rabbits died at 32 ug/kg, 1 died
at 16 ug/kg. Typical clinical picture was severe liver
and kidney injury (histopathologic examination) at doses
approaching LDSO. Have seen histopathology at single

dose of 3 ug/kg. Dose of 500?1000 ug/kg was always fatal;
sometimes took 10?20 days for death to occur.

{Tor parathion, the acute oral LD50 in the rabbit
10,000 ug/kg]

Rat

5 mg in 1:1 corn oil/acetone (5 animals per dose):

Acute LD- female rat
00

If

45 ug/kg
22 ug/kg

ll

male rat

{For parathion, the acute oral LD50 is:

rat(M) 5,000?30,000 ug/kg
rat(F) 1,750?5,000 ug/kgj

 

(1)

Data for parathion taken from ?Evaluation of The Toxicity of
Pesticide Residues in Food?. Joint Expert Committee on
Pesticide Residues. r30 MCH. Rept. No. 

95% confidence limits.

162:: t?

 

Guinea Pig

50 mg in corn oil (5 animals per dose)

 

oral LDSO, female guinea pigs 0.6 ug/kg

:3

[For parathion, the acute oral L050 is 9,300? 

1

32,000 ug/kg for guinea pigs: 

?-Iu



ANALYTICAL METHODS t?
CO

Vapor Phase Chromatography can detect 1 ppm. Need 100 3. E5


for assay. (Method attached.)

Liquid Chromatography can quantitate 1 in a 2 g. sample.

 

Bioassay (see under TOXICOLOGY, Skin Contact, Rabbit).
Can detect the chloracnegen in smaller samples and should
_be used to verify absence or presence of ?dioxin? in 

used in toxicity tests.

DOSE RELATIONSHIP or WHERE PRESEN
AT 10 IN 

 

10 "Dioxin"
mg/kg ug/kg_ ug/kg
1 1,000 0.01
10_ .10,000 0.1
50 50,000 0.5*
100 100,000 1.0
*Equal to the acute oral LD- for the ?dioxin? to female guinea

pigs and 1/6 of the singleooral dose which causes histopathology
in liver of rabbits.

 

limits.

182?"?

 

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5% in DPM 375 M. Intact i 5 be1_y Ex-
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cat?sh. Animals sacri'icadAbradea i 3 I beliy ESLen?lally the same intact 5 ear N0 irritation; no foliiculftf
- Animal died after laSB app 1-
cation.

 

1% in DPH 55? F. Intact 5 belly Same as abnvo, no Hyp?rpla?l?
. v?uIr-v-?? 

 

 

 

 

in DEM 608 H. Intact 6 ear No irrntation, folli?ibs
prominent afta~ last cpbliu
cation. Animal died.

same as above; no Hyperpld?i?

0.25; in DFH 603 M. a Intact 6 beii}

 

 

 

 

0.15 in DFM 932 M. Intact 14 ear Moderate folliculitia;
(l7 ?ayn) only

 

 

 

 

 

 

 

 

 

 

 

 

 

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(17 days) only
1? 

 


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n:

 tion or the? registrations tor useof phenoxy herb1c1de Viz. 2, 4-D, 2, 4, o-T. 

I



. 




C.


2-D

DEFARTMENT - WILMINGTON. DELAWARE 19899 - TELEPHONE: 

November 27, 1967

.I- I - - I "at! I
I I r-?I 4/6 I
. I [2/2 

Dr. Harry Hays, Director

.estici Regulation 71,.  
WashingtonDear Mr. Hayswriting to request that the effective date for possible cancella? 

MCPA, and Silvex, on range and pasture grass be extended for at least one 
year beyond the December 31, 1967 date established for cancellation of
no-residue registrations.

The NACA Industry Task Force on Phenoxy Herbicide Tolerances
has assembled petitions requesting tolerances to cover uses previously
registered on a ?no-residue? basis.. The Department of Agriculture has
cooperated with this group in several ways. Dr. L. L. Danielson was
assigned the function of Department liaison with our group, and has provided
information and?data from USDA sources. Dr. Danielson and other members
of the Department have also consulted with us at various stages in the
preparation of our petition. 

On November 17, representatives of the Task Force met with you
and other USDA officials in the office of Dr. Robert J. Anderson to inform
the group of progress in the preparation of petitions. The status of range
grass and pasture grass as raw agricultural commodities and tolerances
which might be necessary for them were also discussed. Pursuant to the
results of that discussion, we are deleting from our petitions requests for
the 20 tolerances for grasses that had been considered, and requesting
that an extension be granted so that data_necessary for tolerances at a level
higher than 20 may be developed.

Continuation of the registrations for use of phenoxy herbicides on
grass is in the best interests of both the agricultural industry and the public.
The usefulness of these materials in recovering and improving of grass
lands and pasture for livestock production was emphasized by USDA repre-
sentatives at our meeting on November 17. There is no hazard to animals,
as evidenced by the extensive use of these materials for more. than a decade?55" -. . -

- .-5,?mti?'e'd't' 1 8 ifMOGW

Dr. Harry Hays - 2. - November 27, 1967

nor is the presence of residues in meat or milk a regulatory problem.
Evidence from feeding studies concerning residues in meat and milk is
included in the petitions being submitted in support of negligible tolerances
for no-residue uses. 

We have requested a cooperative program with USDA to resolve the
matter of residue tolerances for phenoxy herbicides used on grass, and
where necessary, for meat products} 'We intend to pursue this by a series
of conferences to establish a program that will meet the needs and desires
of the various agencies concerned with the agricultural use of phenoxy .
herbicides, as well as those of the regulatory agencies. A meeting with
Pesticide Regulation Division to further discuss such a program will be
scheduled in the near future. I 

amass MOO

Petitions seeking negligible tolerances for uses of 2,4, S-T,
MCPA, and Silvex presently registered on a no-residue basis (except for
range and pasture grasses) are being submitted by Hazleton Laboratories,'

in behalf of the Industry Task Force. These should be in your hands

 the next week.

.

Very truly yoursDunn, Chairman
industry Task Force on
Phenoxy Herbicide Tolerances

CCD:alc  . .   

cc: Dr. George W. Irving, Administrator

Agricultural Research Service
U. S. D. A. -
 Washington, D. C.

Dr. R. J. Anderson, Deputy Administrator
Regulatory and control Programs
Agricultural Research Service
U.S.D.A.

WashingtonDanielson I i  I  .- - .
Agricultural Research Service -  
Beltsville, Maryland - 

. .- . x: [Cu-,Ir -

 

 

 

Pesticide Petition No. BFOGTE




u- 'r

- .
DEPARTM ENT HEALTH. EDUCATION. AND WELFARE 1 a 6
FOOD AND DRUG ADMINISTRATION

WASHINGTON. 0.0. 2020?

April 13, 1900

Mr. C. L. Dunn, Chairman

NACA Industry Task Force on
Phenoxy Herbicide Tolerances

1155 Fifteenth Street, N.W.

Washington, 20005

Dear Hr. Dunn:



This refers to Pesticide Petition Rb. BF0675 requesting tolerances for
negligible residues of the herbicide silvex at 0.2 on apples, pears,
prunes, rice, and sugarcane. we have completed our review of the petition
and find that we are unable to establish the tolerances as requested for
the reasons listed below.

Our conclusions with regard to the residue data are as follows:

1. The metabolism and degradation of silvex and its ester and salt
formulations are not adequately delineated. Data are also required to
identify and quantitate the major toxic metabolites which may be present
in plants at normal harvest; of particular concern are conjugates of
silvsx such as those reported by Meagher, J. ?53. Food Chem. l1,
37-1-6 (1966). If such metabolites are found to constitute a significant
residue at harvest, an analytical method for their determination will be
required. 

 

2. The methods of analysis outlined in the Pesticide Analytical Manual,
Vol. I, 6.11, 6.21, and 6.33CA), would not be expected to deter?
mine the metabolites mentioned above.

3. The present residue data are not sufficient to conclude whether the
proposed tolerances are adequate to cover all toxic residues, including
the metabolites mentioned above. In addition, residue data (including
toxic metabolites if found present) are required for apples reflecting
the concentrated spray application; (b)-for dried prunes; for rice
bran and straw; for sugarcane bagasse and sugarcane products (syrup,
molasses, sugar) derived from cane bearing residues of 0.2 ppm. (The
present study of residues in refined products does not indicate whether
residues were found on the cane from which the products were derived).
The application rate and PHI used in the present residue data for prunes
should be furnished. The additional residue data should represent appli-
cation of the most persistent esters or salts of silvex. If residues are

 .



(Mr. Ce L.  2

concentrated in any of the above processed foods, appropriate food addi-
tive tolerances should be proposed.

4. Depending on the level of residues found in livestock feed items
(rice bran, rice straw, bagasse, and molasses), we may need additional
tissue storage data as a basis for concluding whether or not residues
would transfer to meat and milk.

We are unable to draw final conclusions with regard to the adequacy or
the toxicity data until the necessary information is received. You may
amend the petition or you may withdraw it and refile when the necessary
information is available. Otherwise it may be necessary to establish a
tolerance or zero for residues or silvex on the crops in the petition.
Please let us have your reply within 30 days.

guess/woo

Sincerely yours,

William Stokes

Petitions Control Branch
bureau 01 Science

co Pesticides Regulation Division
ARE - USDA



 .. .. . . 

THE MADISON BUILDING

Cor?ed to:
I A I A n55 STREET. MW.4 

1? IAng WASHINGTON c. 20005 1.40%"
File I I . . . 
AGRICULTURAL
CHEMICALS

ASSOMATION

 

Tllophnno: 295-l585
Area Coda 202

 

?y

.hUJ'll, 1968 c5

2 

2; on

c: 

F0

hm. Drew H. Baher, Jr. CD 
Petitions Control Branch n- 2;
Bureau of Science 

Department of Health, Education, and Welfare
Food and Drug Administration
Washington, D. C. 2020h

Dear Mr; Baker:

I am writing in regard to Pesticide Petition Nos. 8FC669 
8F067O 8s0675 (Silvex); and 830676 (tern). Letters detailing ob-
Jections, primarily concerned with questions concerning metabolic products,
were sent to us in April, 1968. On April 25 representatives of our Task

Force met with you, Mr. Cummings, and Ramsey to discuss these objections.

At the meeting we stated that we would prepare an additional section
for the petition reviewing the scientific literature with respect to metabolism
of phenoxy herbicides and emphasizing the application to residues. Work on
that project was started shortly after our meeting, and I wish to report on
its status at.this time.

Publications dealing with the metabolism of phenoxy herbicides in
plants and animals as well as recent residue studies have been screened.
From this material, about 60 references and review articles have been
selected as being pertinent to this review. Documents have been obtained,
examined in detail, and information tabulated. In some cases authors have
been written or called for additional information or clarification of their
research findings. The writing of various portions is currently in progress,
and it-is estimated that the draft will be completed in about two weeks.
Revision and reproduction of this somewhat bulky document will require an
additional several weeks, and we are aiming at submission of the document

'by about August 15. 

- 

IKIEWSLE 

JUL 151968

BIOPRODUCTS DEPT.

arm

-- .- 

"i?ass

 ir. Drew M. Baker, Jr. - 2 -

July 11, 1968

'An outline of the section that is being prepared is as follows:

Table of Contents
Summary Statement
Glossary of Terms

I. Metabolism in Plants
A. Introduction
B. Tabulation of Conditions Used in Letabolic
Studies in Plants?_
C. Metabolism of 2,h-D
1. Hydrolysis of Esters and Salts
2. Decarboxylation
3. Conjugation
Formation of Water Soluble Unknown 1
Formation of Water Soluble Unknown 3
h. Transformation to a Different Ether Soluble
Compound 
D. Metabolism of 2,h,5-T and 
E. Letabolism of MCPA and MCPB
F. Metabolism of Silvex 



II. Metabolism in Animals
Recent Residue Studies by Newer Methods

IV. Significance of Metabolites as Residues in Foods
and Feed

Please let me know if there are further questions about our activity
in the execution of our commitment to prepare this additional information

discussed at our April 25 meeting.
Very truly yours,



C. L. Dunn,.Chairman

NACA Industry Task Force on

Phenoxy Herbicide Tolerances
..CDD:mea

cc: USDA Pesticides Regulation Division

CHLORACNE

Dow EXPERIENCE :TCBD 2;
O1

 

15291"

35391 

'313 330W

3811 UBNIV13 ANVN ?83A11 '5

166_

38 SNIWHVSIU 'h


HO Oi SHVSA 01 - Nail-9N01

W. 1 



3Hi ?0 HBHLO

GNV MOVE 3H1 Oi AVW NO 18315 

'savaHx3v1a 330w Lna 'auav EBVNEEL 3x11 

N1 3N3VH01H3

DON EXPERIENCE 1964 TO PRESENT

1. ABOUT 100 CASES.

2. ABOUT 60 IN TCP INCIDENT DESCRIBED BELOH.

3. MOST OF REMAINDER IN DOWICIDES PRODUCTION.

4. SOME CASES IN LABORATORY RESEARCH.

THREE EXTREMELY SEVERE CASES, ABOUT FIFTEEN SEVERE

5.
CASES, ABOUT THREE CASES, ABOUT FOUR 




6. MOST CASES ARE MILO, MANY WERE FOUND IN HEALTH
INVENTORY ACTIVITY BY 

MW

18383

1934-1936

1930?s

SIGNIFICANT CASES -- DON

DOWICIDE PRODUCTION -- SEVERE OUTBREAKS AMONG
EMPLOYEESSODIUM 

CHLOROPHENATE



LABORATORY CASE 



Active

DIPHENYLENE OXIDE: ALSO DIBENZOFURAN

1991



1966

D. D. IRISH AND E. M.-ADAMS
PUBLISHED PAPER ON RABBIT EAR TEST
FOR PREDICTING ACNEGENS: STILL THE
MOST SENSITIVE TEST AVAILABLE.

DON CUSTOMERS CONTRACTED CHLORACNE..
FROM THEN DONICIDE 3 (Now

DONICIDE 31 a 32, HIGH PURITY THAN
DONICIDE 3).

OH cl



or

c/

MIXTURE 0F 4- AND 
PHENOL

PRODUCT AND PROCESS WERE CLEAN
EXCEPT FOR TAR.

Mn?

1942

1950?s
TO 1960?s

RESEARCH LAB ONE SEVERE

CASE FROM RECYCLING TCP RESIDUE IN

GLYCOL SOLVENT. AN MOLECULE

NAS IDENTIFIED BY IR AND HYPOTHESIZED \yn 
AS CULPRIT, BUT NOT CONVICTED BY Q57)
THE EVIDENCEPRODUCTION -- AT LEAST ONE
SEVERE CASE, SOME MILD CASES. DON

oz. 14-94 I mun

TERNINATED PRODUCTION BECAUSE IT 
LON-VOLUNE. LOW-PROFIT ITEM, NOT -
NORTH THE COST OF RE-ENGINEERING TO
PREVENT FURTHER INUURY.

Cl4_6 Actlve
0
C18 Inactive

1963-1964

CHEMICAL-PHYSICS LAB -: SEVERAL 
AT LEAST ONE SEVERE, FROM RECYCLING
CAUSTIC INSOLUBLE OIL IN ALCOHOLIC
HYDROLYSIS OF TETRACHLOROSENZENE.

AT LEAST ONE EXPLOSION OCCURRED.

WESTERN DIVISION TORDON PROCESS 
FOUR MILD CASES. RABBIT TESTS FAILED:
SUGGESTING NEW FAMILY OF ACNEGENS IN

CHLORINATED PYRIDINE CHEMISTRY.

NH
C1- C1

COOH

ACID

 

*0

1966

.. \2
. - 
DOH CORNING RESEARCH LAB CHEMIST (QO
. 
HEATED TCP FOR RESEARCH PURPOSE - 
UNRELATED TO DON PROCESSES, ISOLATED 
TCBD AS AN UNKNOHN BUT INTERESTING-
ECAME ONE OF OUR BETTER

   
  

BY-RRODUCT

   

SOURCES

CASUAL DISPOSAL OF LAB HASTES
RESULTED IN ONE MILD CASE -- THE


ZLILQZJ M?u

y:


Q55 .1
OD

;i83l adlM 01 109
3M 3H1 AH NV313 SVM 



'aHl NI 33315 338Hl 1v N338 GVH
HBIH GNV uaxaa
BQIDIMOQ 'N0113naoaa
BGIDIMOU.01 031v1aa sasva N31 inoav
- vavwva MOO Laos

996I

.1966

1968

UNIVERSITY OF IOWA 2,3,7.8?
TETRABROMODIBENZO-P-DIOXIN APPEARS TO
BE AS POTENT AS TCBD. AT LEAST ONE
SEVERE CASE: DID NOT RELATE IT TO HIS
RESEARCH UNTIL ASKED FOR A SAMPLE AND

DESCRIBED THE PROBLEMONE MILD CASE RESULTED FROM 
DICHLOROPHENOL TARS FROM 3A9 BUILDING.
THE TAR HAS USED IN A PUMP TESTING

SET UP AT 562 BUILDING. POSSIBLE
HAZARD HAD BEEN MENTIONED BUT NOT
EMPHASIZED TO USERS. VICTIM DISMANTLED

CONTAMINATED PUMPS WITH NO PRECAUTIONS.

Pi.
"Cb

1930's

OTHER EXPERIENCE

MANY SEVERE CASES INCLUDING LIVER INJURY
RESULTED FROM CHLORINATEDINAPHTHALENES:
THREE CHLORINE EQUIVALENTS OR MORE.

C13

FRENCH MANY SEVERE CASES. REVISEO -
PROCESS. WERE VERY SECRETIVE. USEU I
MOUSE TEST IN PLACE OF APPEARED
TO BE IGNORANT OF MANY IMPORTANT ASPECTS
OF PROBLEMS.



18301

1955

1957

196?

GERMAN SEVERE CASES INCLUDING SOME
SUSPECTED FATALITIES. SHUT 
RESEARCHED NEW PROCESS.-ISHOWED AGAIN
THAT RABBIT EAR AND LIVER ARE MOST
SENSITIVE TESTS.

SENT A REMARKABLE LETTER TO ALL KNONN -
PRODUCERS OR TCP DESCRIBING PROCESS
PARAMETERS NECESSARY TO AVOID
CHLORACNE HAZARD LETTER WAS FILED
AND 

GERMANS WERE QUITE KNOWLEDGEABLE 
VERY COOPERATIVE. DOW BOUGHT PROCESS
INFORMATION.





pa

yd
C9
C3
,bo~

 1946

1952

THE ANILINE MOUNTAIN STORY -- 199 BUILDING

BATCH PROCESS START UP: RAN UNTIL

1952.

BEGAN CONTINUOUS COIL REACTOR
PROCESS.

C. I. OIL AT 10% SEVERE
FOLLICULITIS.

EXPERIMENTED NITH 73% AND 24%
CAUSTIC T0 REPLACE FLAKEDIFFERENCE IN SEVERITY BETWEEN
FLAKE PRODUCED AND ZNZ CAUSTIC

PRODUCED OILS.

2.2. 

183031

 1953

JULY

INOVEMBER

NEN EQUIPMENT START 
PLUGGED LINES PROBABLY
PRODUCED MORE EXPOSURE.

PUSHED PROCESS TO INCREASE
PRODUCTION, RAISED
TEMPERATURE FROM 195? TO
205?225? C. MADE MORE OIL.
BEGAN SAMPLING IT.

MW

15304.

4-0

FEBRUARY

APRIL 1

APRIL 15

APRIL 20

MAY

JULY

SUPERVISORS, LAB MAN AND
OPERATORS WITH CHLORACNE.

PHOENIX SPRINKLER MEN MOVED
IN.

REDUCED EXPOSURE BY EOUIP-
MENT AND PROCEDURE CHANGES.

c. r. OIL AT 0.25% KILLED
RABBITS 

PHOENIX MEN MOVED OUT.

BLILQL Man

O. I. OIL AT 0.25% KILLED
RABBITS.

c. I. OIL AT 10% SEVERE
FOLLICULITIS.

PHOENIX MEN WITH CHLORACNE.

16305

MID'lg?q MANAGEMENT ASKED Two OUESTIONS
- OF MEDICAL, INDUSTRIAL HYGIENE

AND SAFETY:

(1) CAN PLANT BE OPERATED
SAFELY AT 

(2) WHAT TO ENSURE


ANSWER TO (1) WAS YES.

ANSWER TO (2) COST MANY AND
MUCH MANPOWER.

18306

PROCESS CONTINUED 1964?1966 NITH NO NEH
CASES OR ADVERSE EFFECTS ON OLD ONES,

TNO CASES IN 1966 -- DID NOT FOLLOW
RULES.

M00

TBILSLI

DISPOSITION OF 199 BUILDING

PRIZE WINNING SUGGESTION 

IT AND MAKE A SKI RESORT,
CALL IT ANILINE 

DISMANTLED.IN 1968 -- NO INJURIES.

1 Mon

EBTLQA

1-63

{x

8.

NEW PLANT -- BUILDING

 H- 7 PM 77:01) moans

- 
c. I. OIL 30-50 PPM TCBD 
PRODUCT 'ALwAvs LESS THAN

4 a; #97 

2
Id.

WIPE TESTS CLEAN ?3
U)


C0
03

pt;

?229
CD
CD

 COMPOUNDS

0

KNOWN ACNEGENS

ACTIVITY



C13 

DATA NOT towcuusxvs

UNKNOWN ACTIVITY

UNKNOWN ACTIVITY

SOME VERY ACTIVE-

NOT ACTIVE

SOME ACTIVITY
NOT ACTIVE

SOME ACTIVITY

T494 1 may


(3'3
CD
In. 
CD

MATERIALS POSSESSING UNEXPLAINED CHLORACNE ACTIVITY

on . . 1
c1 c1 SOME MIDLAND BATCHES ARE
MILDLY ACTIVE
MANY CANADIAN.BATCHES ARE
cl cl MILDLY REACTIVE
c1 -
OH 
c1 c1
1
DOWICIDE 
C1 c1 ALL SLUDGES ARE ACTIVE Egg
1


.cn
- 0Na :3
ALL DECOMPOSITIONS CD
ARE ACTIVE 
11-5 A
on . .
HEAT SOME HAVE ACTIVITY

c13_5

18311.

COMPOUNDS KNONN TO BE INACTIVE

CARBON TETRACHLORIDE


AT LEAST ACTIVE



UCTACHLOROBENZODIOXIN

DON TETRACHLOROBENZENE

5-3

FIE

5.,






DOWICIDE 9
Q-CHLOR0-2-CYCLOPENTYL PHENOL

ACCUSED IN LITERATURE:

15312



PLATITUOES

AVOID TEMPERATURES GREATER THAN 150c C.I
BEWARE 0F.TARS AND OILS

LABORATORY SCALE IS HAZARDOUS

USE RABBIT TEST EARLY

COMMUNICATE INDUSTRIAL HYGIENE, MEDICAL





3 

3?


4:5)

-- 

$2

MIDLAND, Michigan

4m THE DOW CHEMICAL COMPANY 
1 t}
i 
up

August 16, 1966 2;

co

cn

o. A. Highhill
2 Plant
489 Building

cc: E. C. Staehling, Org. Chem. Prodn., 258 Building
C. Amstutz, Herbicide Section, M41 Building
I. Chase, Products, nag Building
B. Holder, M.D., Medical Department, 607 Building
. w. Wilson, Safety Department, #01 Building
_53 Rowe, Biochemical Research Laboratory





I would like samples from your first run with 2,u,5?
trichlorophenol in the direct ester process. The animal
test will detect the presence of chloracne-producing
compounds. -

Please send me a sample of the final product and any
intermediates that you think should be tested. If
activity is detected, the intermediate may tell us

_where the trouble starts.

Because of the concern about chloracne, I think we should
run animal tests on esters for a time to prove
the lack of hazard, even if the first tests are negative.

twee:
L. G. Silverstein
Biochemical Research Laboratory

1701 Building

.a



 

    

 

7

xm?W.
i' OCCUPATIONAL

DISEASES
Maw, cm (Hi 

. 

A Guide
To Their Recognition

Rewritten and Enlarged Edition of

 

()cruputiun Hnumlx and Diagnostic Signs

 

 

OF HEALTH.
AND WELFARE

Public ?with 

W. M. GAFAFER, D. Sc., Editor

Division of Occupational Health

28 JNAL DISEASES

may be. damaged by maceration, mechanical or chemical trauma, and certain
internal diseases. Lipid-soluble and nonionizable (organic) substances are
able to bypass the barrier layer and enter by way of the transfollicular path-
way. Cases, except for carbon monoxide, are readily exchanged through
the epidermis.

Protection against actinic stimuli is afiorded by increased melanin pro-
duction and a concomitant thickening of the keratin layer. An antimicrobial
action is attributed to certain fatty acids in the surface lipid ?lm, and the
subcorneal barrier layer also offers some protection against microorganisms.

Primary Irritants

Primary irritation causes most of the cases of contact dermatitis in in-
dustry. A primary irritant is an agent which will cause dermatitis by direct
action on the normal skin at the site of contact if it is permitted to act in
sullicicnt intensity or quantity for a suf?cient length of time. Thus, the
normal skin will almost always react to a primary irritant if the necessary
conditions are present.

lrritants can act in several ways to injure the skin. Several examples have
already been given, such as removal of lipid ?lm, denaturation of keratin,
and interference with the subcorneal barrier layer. Other examples ofi'pri-
mary irritation include dehydration by inorganic acids, and
alkalis; protein precipitation by heavy metal salts and tanning agents; and
oxidation by lilt??Cltt?S, chlorine compounds, and per- salts.

Sensitizers

Ordinarily, no men- than 20 percent of contact dermatitis in industry
is caused by allergens. HoweVer, certain of the strong sensitizers may
affect many workers. Almost any chemical can be a sensitizer, but the mode
of action usually differs from that exhibited by primary irritants. The
cutaneous sensitizer, therefore, does not necessarily cause demonstrable
cutaneous change on first contact, but may effect such speci?c changes in the
skin so that after 5 to days or more, further contact on 'he same or other
parts of the body may cause dermatitis. 

The difl'erence between the irritant and the sensitizer, therefore, is generally
a matter of time as well as mode of action. The irritant will usually act
within a matter of minutes to a few hours, whereas the sensitizer requires
at least 5_ to 7 days because sensitization must build up during the period
of incubation. Low-grade irritants such as soap may require prolonged or
repeated contact before a dermatitis appears, and this prolonged reaction
ime may be confused with allergic incubation.

iccondory E?ccts

After the cutaneous defenses have been broken down and a contact der-
ualitis is present, the oozing or fissured surface offers ideal conditions for
be cunt-misc and growth of bacteria. Thus, it is not unusual for a contact

co 
pus

.31

 

OCCUPATIONAL DERMATOSES 29

dermatitis to he secondarily infected. Another secondary effect commonly
seen in contact dermatitis is over-treatment by irritating or 

medication.

Diagnosis

Diagnosing an occupational dermatosis is generally contingent upon satis-
fying certain criteria. The disease should look like a contact dermatitis or
one of the other clinical types of occupational dermatoses. it should be
located on sites of exposure, and the time of appearance of the eruption as
well as periods of rcmisson and exacerbation should correlate with the history
of work exposures. ?'hen the patient does not get well following complete
withdrawal from the suspected contact agent, stimuli of nonoccupational
origin should be reinvestigated. Multiple factors not associated with the
occupation can perpetuate a chrOnic dermatosis.

Patch tests are helpful in differentiating between a primary irritation
dermatitis and an allergic contact dermatitis. A nonirritating concentrav
tion of the suspected allergen is applied to the normal skin of a patient for
24 to 48 hours in an attempt to reproduce an eczematous dermatitis beneath

the covering patch.

Classi?cation 0/ Lesions

The clinical lesions seen among occupational exposures are multiple,
varying from the mildest erythema to lesions of neoplastic nature. How-
ever, occupational skin disease can be classi?ed as follows:

(1) Acute contact eczematous dermatitis characterized by erythema,
edema, papules, vesicles, or bullae, crusts and finally desquamation. Tthe

eHects are generally the result of contact with either a primary or a 

sensitizer, or with both.
(2) Chronic contact eczematous dermatitis characterized by erythema,

lichenification, and ?ssuring of the skin, usually resulting from contact with
dehydrators, fat solvents, soaps, and detergents. I

(3) Folliculilis and acncform types, including chloracne, characterized by
plugged sebaceous follicles and suppurative lesions. Chloracne also shows
numerous straw-colored cystic lesions. These forms of occupational derma-
toses are usually caused by contact with oils, tars, waxes, and Certain
chlorinated hydrocarbons. I

(4) Neoplastic {benign and malignantl types, characterized by keratoses,
papiliomata and epitheliomata of the exposed areas, usually caused by petro-
leum products, coal tar and certain derivatives, sunlight, and 
radiation. I

(5) Pigmentary disturbances characterized by increase or decrease of pig-

ment in the epidcmtis. Increased pigmentation can result from contact with

Mitt]





 

o-Dichlorobenzene workers
Dry cleaners

Dye makers

Fumigant workers
Creasemakcrs

CHEMICAL HAZARDS 117

Varnish makers

Varnish remover workers
Wax makers

Wood preservative workers
Wool processors

 
 
  
 
  
 
 
  
 
  

 

 
  
 
 
 
 
 

   
  
 
    
     

Cum makers  
Heat transfer workers Deodorant makers In."

Hide processors p-Dichlorohenzene workers 

Insecticide workers Disinfectant workers ?3

Lacquerers Drug makers 
Lacquer workers Dye. makers 
Metal degreasers Workers DJ
Metal polish makers Moth hall makers 01

Organic chemical Soil iumigalors *3

Paint remuvcr workers Trio/tinrohennrm's C'n
Paint workers Dye makers
Polishing compound makers Electric equipment makers 
Resin makers Heat transfer workers 
I: Rubber makers Insecticide workers  
Solvent workers Lubricant makers  I 

Stainers Trichlorohenzcne workers  

Stain makers chacl'ilorobenzenc 

   
 
 
 
  
 
 
 
 
 
  
 
 
 
 
  
 
 
 
  
     

Sullur processors l-?ungieitle workers

Tannery workers llexach[orolu-nzene workers

Tar makers Organic chemical 
Tar remover workers

Termite exterminator Workers

Set-ti disinfectan

References .

IL L.: Ime. v. K.: OVEN. Iltn'LE. ll. IL. AND It. 
Toxirity ol paradii-hi"ruin-menu: determinations on experimental animal- and human
:ul?jt?t'lb. Arch. ?cull/t liili, l?ISh.

H. L.: Itqu. V. ores, IL. .4in H. M.:
Toxicity of o-dichlorobenzene; studies on animals and industrial i-xpi-rin-nr-z 

Arch. Imiust. Hr'ulth 17: 180. i953.
(48) Chlorinated anil 
Harm/u] Efforts

Local Prolonged c0ntact with fume or the cold wax lemls on exposed skin
to corneilones, sebaceous and pustules, known as chlorac-ne.

Routes 0/ Fairy inhalation of fume or vapor; pr-rt-ntanmms absorption
of liquid.

Systemic Acute or chronic exposure can produre varying ilegrees of liver

damage depending on amount of chlorine in compound ami 

 

 

   
  
 
   

118 

DISEASES

state of liver. include jaundice, anorexia, nausea, indigestion,

abdominal pains, and edema. Death
I has occurred.

Special Diagnostic Test
None.

Recommended Threshold Limit

Cltloroclt'pltenyl (42 percent chlorine),

from acute yellow atrophy of liver

1 milligram per cubic meter of air.

Should be reduced when also absorbed pcrcutaneously.

Cltlurmlt'pltena'l {5-1 percent chlorine)

. 0.5 milligram per cubic meter of

air. Should be reduced when also absorbed 
Chlorinated clip/tenyl oxide, 0.5 milligram per cubic meter of air.

0.5 milligram per cubic meter of air. Should be
reduced when also absorbed percutaneonsly.

5 milligrams per cubic meter of air. Should be re.
duced when also absorbed percutaneously.

Potential Occupational Exposures
Aniline dye makers
Cable coaters
Carbon removers
Chlorinated diphenyl workers
Condenser impregnators
Crankcasc oil additive makers
Dye makers
Electric equipment makers
Electricians
Electroplaters
Flameprooiers

.. .

Cum processors

Heat transfer workers

Herbicide workers
makers

insecticide workers

insect prool'ers

Lacquerers

Lacquer makers

Light ?xture makers

Machinists

Metal ClL?gl?E?th?S

Reference

 

:3 arocltlar. Med. Assoc. 154; 1417,1954

 

macs. J. ALBUM. J. mo swarm, a. L.: Chloracne from an unusual exposure

Mineral oil processors
Moisture prooiers
Paint makers

Paper treatcrs
Petroleum re?nery workers
Plasticizer makers
Plastic makers

Rayon makers

Resin makers

Rubber workers
Solvent workers
Stainers

Stain makers

Textile llnnu?pruofcrs
Transformer workers
lipper cylinder uil tuakt-rs
Varnish makers
Vegetable oil processors
?iax makch

Wire coaters

Woad preserveHarmful E?ects

Local Extreme irritation of
sion of teeth.

Route 0] Entry Inhalation of
Systemic Acute respiratory
pain. and 
nia. and pulmonary edema ma

Special Diagnostic Test

None.

Recommended Threshold .

1 part per million parts of a
of air.

Potential Occupational Ea

Aerosol propellant makers
Alkali salt makers
Aluminum puri?ers
Benzene bexacbloride maker
Bleachers
Bleaching powder makers
Bromine makers
Broom makers
Carpet makers
Chemical 
Chloride of lime makers
Chlorinated solvent makers
Chlorine workers
Color makers
DDT makers
Disinfectant makers
Dye makers
Ethylene glycol makers
Ethylene oxide makers
Flour bleachers
'Freon makers
Gasoline additive workers
Cold extractors
makers
iodine makers
lron dctinners
lron dczinkers

..
?s

. - 'i 
?minim? 0- 
- a - 

4



to



l? ?hicides, or weed killers, may be classi?ed as pesticide chemicals. The:
cat .l plants on contact, or they can be translocated; that is. absorbed 
one part of the plant and carried to other parts where they exert their print-trW
toxic ell'ect. Most of the commonly used herbicide-55ammunium 
dalapon, phenoxyacctic acid derivatives. carbatnate derivatives petroleunu
oils. sodium borate, Crag" herbicide??have a low toxicity and liave caused
little dif?culty among users.

borne pose more serious problems; for example. the central
nervous system effects of maleic hydrazide or the methemoglobinemia and
central nervous system depression of sodium chlorate. l?entaehlorophenol, a
metabolic stimulant. has been responsible for several deaths because of
hyperthermia. Amino triazol has produced cancer in experimental animals
but there have been no untoward e?ects reported in man. 

llerl-it'ides with cutaneous effects include. trichloroacetic acid, a' corrosive
irritant of the skin and mucous membranes; maleic hydrazide, a producer
of ?allergtc contact dermatitis; pentachlorophenol. a producer of a primary
Irritant type of contact dermatitis; and creosote, a primary irritant and
photosensitizer.

Reference may be made to chemicals in the section on Chemical Hazards
for the toxicity of the following herbicides: Arsenic trioxide and sodium
arsenate lsee Arsenic). copper sulfate (see Copper and Compounds), creosote
compounds lsee Cresol and Phenol), dittitrophenols lsec Dinitrophenol),
kercsinc. and acetate (see Mercury and Compounds).

'l'hreshold limit values. in milligrams per cubic meter of air, have been
recommended for the following;r herbicides as shown.

Ammonium sulfatnate )5 mg, per cu. m.

Crag!" herbicide )5 mg. per cu. m.

2.4?l) f2,-l-dichlorophenoxyaeetic acid)- 10 mg. per cu. m.

(PCP) 0.5 mg. per cu. 

l?henylmercuric acetate (PMA) 0.0] mg. per CL. (for organic
mercury)

l2,4,5-tritrhlorophenoxvacetic
acid).

10 mg. per cu. m. (tentative)

'Shouhl be reduced when also absorbed 

Furnigants

Fumigants are pesticides which may be applied in the solid, liquid, or"
gaseous state. A combination of high volatility with high pest toxicity is
generally desired; however, compounds with low Volatility may be preferred

for sod fumtgatton. The possibility of excessive exposures exists wherever

 

 

[Hunts-H? .

houses. barns. :llillS, mills. lrt-ieht Pitt". and greenhouses.

Each of the following has found use as a funny .it. Because 1

they have other industrial appliratinns as uell. they are discussed in.lividua1ly .

..

in the section on (Chemical Hazards.

Methyl Bromide lsee Bromine and
Compounds)

Methylene Chloride

Acrylonitrilc
Carbon Disnl?de

Carbon 'l?etrachloride

p-Dichlorobenzene (see Chlori- Methyl Formate
nated Benzenes) Naphthalene
Dioxane Perchloroethylenc

Propylene Dichloride
Sulfur Dioxide

Ethylene Dibromide
Ethylene Dichloride

Ethylene Oxide Tetrachloroethane
Hydrogen Cyanide Trichloroethylene
References

J. ornttut. w. r.t.t.tnr'r. J. AND WOLFE, H. R.: Exposure to para-
thion. Measurement by blood level and urinary p-uitrophenol excretion.
Arch. Entiron. Health 3: 476. 106].

ASSIHZIATIHV or CONTROL orrtcuLs: Pesticide Chrmiutls 0i-
?cial Compendium. A. B. llt-aey. University Post Of?ce, College Park. 1962.
Loose-leaf senice.

CHMMITTEE on PESTICIDES: Outlines of information on pesticides.
fungicides. I. Am. Med. ulnar. LET: 237. I955.

ox Toxtanorz?r: Occupational dit-ldriu poisoning.
172: 2077. 

Comm-nuance onsets: am: SERVICE: Clinical nit-moramta on
economic poisons. l?ub. Health Sentim- l?uh. No. 476. Government Printing Ollie",
Washington. 1936.

amount: or AND NATIONAL nest-1mm! NtTtoN ACADEMY
Vol. 3. Insecticides. Vol. 5. Fungicides. W. B.

Part 1. Agricultural

I. Am. Med. Assoc.

01' Handbook of Toxicology.
Saunden (.20.. Philadelphia, 195?).

DURHAM, w. E: canes. T. 9., no HATES. w. 1. 1a.: Paralytic and related e??erts of
certain organic phosphorus compounds. Arch. Indust. It'rahh 13: 326. 1956.

ut'nunt, arm w. 1., Ureanir phosphorus poisoning and its therapy;
with special reference to modes of action and compounds that reactivate inhibited
cholinesterase. Arch. Environ. Health 5-. 1962.

FREMI. D. E. II. iEDtToa): f?estiriife Index. College Science Publishers, State College.
l?thl.

CHNES. T. The acute toxicity of pesticides to rota.
2: 88. 1960.

cows. It.
Autsrit?an i'vanamitl Co,, New York. l?Hnll.

ttn'tzs, w. Jtt.: The toxicity of dit-ltlrin to man; report on a survey. Boil. 
'20: )950.

HAYES, W. 1.. JILI


:ta-o-H O-tio?t"

Tort'ml. Appf. 

ll. tum 51Hrt?t?R. C. tI.: Toxit'ot'ogiruf on 

Pesticides in rriation to pubiic health. Ann. Rev. Entomot'. 5:

useat Hill?

1-.

 

 

 

i: t.  ..


Section 

 

Dow 537307

.11. 

 

 

 



Petition for the Establishment of
Tolerances for the Pesticide Chemical,
2,h,S?TTichloroPhenoxyacetic Acid

on Raw Agricultural Commodities

SECTION 

Full Reports of Investigations Made with
Respect to the Safety of the Pesticide

Chemical, 2,h,5~Trichlorphenoxyacetic Acid

December 8, 1967

8081.89 MOO

- 15323

Acute Studies

Oral Administration RatsZ Mice, Guinea Pigs, and Chicks

The acute oral LD50 of 2,h,5?trichlorophenoxyacetic acid

for young adult male and female albino rats, mice, and guinea pigs and

 

for young chicks, as reported by Rowe, et al. (1), was as follows: (3
Species LD50 Confidence Limits Slope 
(mg/kg) to

Rats 500 39]. -6?40 1 .  
(23
Mice 389 ens?619 1. 80 :0
Guinea Pigs 381 3074:72 l. 21
Chicks 310 21.14456 1.1m

The material was administered as a 3.0 or 10% suspension in
olive oil. Gross toxic signs consisted of ataxia and myotonia, particu?

larly at the higher doses.

Oral Administration - Dogs

According to Drill and Hiratzka (2), the acute oral LD5Q of
2,h,S-trichlorophenoxyacetic acid for adult mongrel dogs is probably

in the range of, but higher than, 100 mg/kg of body weight.

Only 25 mortality occur?red "t this level. Gross toxic signs :consi'sted of
anorexia,.body weight loss, slight or moderate stiffness in 
legs, and ataxia. Histologic changes included some necrosis and
inflammation of the intestinal mucosa, moderate diffuse hepatic necrosis,

and a mild degree of renal tubular degeneration.

18323

C2
Acute Oral Administration Rats, Mice. Guinea Pigs; RabbitsL_Chicks,

and Steers

Rowe and Hymas (3) reported the acute toxicity of various for-

mulations of 2,h,5?trichlorophenoxyacetic acid and mixtures of 2,h-

Hie reported LD values

dichlorophenoxyacetic acid and 2,h,5-T. 50

and/or range of toxicity are as follows:

Esteron 2h5 (old formulation containing 33.3% isoPrOPyl ester of 2,h,5?T

and 12.1% mixed amyl esters of 2,h,5?T)

Adult female albino rats Estimated L050 of 1000 mg/kg of

body weight (range 300?2000 mg/kg).

0181.89 MOO

Esteron 2h5 (new formulation containing 65.3% mono?, di-, tripropylene

glycol butyl ether esters of 2,h,5-T)

Adult male albino rats - Estimated LDBO 800 mg/kg of body weight

(range 600?1000 mg/kg).

Isopropyl ester of 2,h,5-T

Male and female albino rats - LDSO A95 mg/kg of body weight

(confidence limits h20-58h mg/kg).

Male albino mice - LD50 551 mg/kg of body weight (confidence

limits 380-799 mg/kg)
Male guinea pigs - LDSO hh9 mg/kg of body weight (confidence
limits 

Mixed Butyl esters of 2,h,5?T 

Male rats - LD50 h81 mg/kg of body weight (confidence limit

313-739 ms/kg).

18324

C3

Male rabbits - Estimated LDgo 712 mg/kg of body weight (range
500-1000 mg/kg).
Male mice - LDSQ 9&0 mg/kg of body weight (confidence limits

67h-1312 mg/kg).

Guinea Pigs - Estimated LD5O 750 mg/kg of body weight (range

500-1000 mg/kg).

Mixed Amyl ester of 2,h,5-T

Female rats - Estimated L050 750 mg/kg of body weight (range

500-1000 mg/kg).

Brush Killer (52.2% butyl esters of 2,h,5?T)

. Female rats - LDSO 1200 mg/kg of body weight (confidence limits

783~1850 ms/ke).
Female guinea pigs - LDBO mg/kg of body weight (confidence

limits 875-2290 mg/kg).

Female mice - LD50 1230 mg/kg of body weight (confidence limits
938-1620 mg/kg). 

Male rabbits - LD50 8H9 mg/kg of body weight (confidence limits


Chicks - LD50 2000 mg/kg of body weight (confidence limits

1350-2960 mg/kg).

Brush Killer 50-50 (formulation containing 27.2% butyl esters of and
26.5% butyl esters of . .?iat wigw a.
I 

Female rats - LDBO 1070 mg/kg of body weight (confidence linits

700?1650 mg/kg).

ITELEQ MOO



is;

.

J.

ch
Female guinea pigs LDSO llfO mg/kg of body weight (confidence
limits 820?1630 mg/kg).
--Female mice LDSO 800 mg/kg oi body weight (confidence limits
62h?1070 mg/kg).
Female rabbits Estimated LD50 lh20 mg/kg of body weight (range
500-2000 mg/kg).
Chicks - L050 #000 mg/kg of body weight (confidence limits
2700?5900 mg/kg).
Esteron Brush Killer (old formulation containing 25.6% isopropyl ester of
2,h-D and 2h.h% isopropyl esters of 2,h,5?T)

Male rats Estimated LD5O 1000 mg/kg of body weight (range

ETELQS Moo

300-3000 mg/kg).
Esteron Brush Killer (new formulation containing mono-, di-, and triproPylene
glycol butyl ether esters of 2,h-D 3u.8% and 2,h,5-T 33.0%)

Male and female rats - LD50 860 mg/kg of body weight (confidence

limits 800?930 mg/kg).

Male guinea pigs LDSO 1220 mg/kg of body weight (confidence

limits louo-lu30 mg/kg).
Female guinea pigs LD5O 1600 mg/kg of body weight-(confidence

limits 1390?18h0 mg/kg).

Male and female rabbits LD50 960 mg/kg of body weight (confidence

limits 790-1160 mg/kg).
Chicks Estimated LD50 2000 mg/kg of body weight (range 1000?




3000 mg/kgweight.

1

Steers - Acute oral LD50 greater than I000 mg/kg of be

163251




Application Rabbits

l-According to a report by the Dow Chemical Company (A), a 10%
solution of 2,h,5?T in propylene glycol produced severe conjunctivitis

and moderate corneal damage which persisted for longer than five days.

Acute Oral Administration - Rats; Acute Dermal and Application-Rabbits

8
The results of acute studies on Nu Amine a formulation gig I
containing 1,3-propylene diamine salts (11.95%) and 2,h,5-T it?
(11.85%) was reported to Diamond Alkali Company February 23, 1962 by it?

Cd

Hazleton Laboratories, Inc. (5).
The acute oral LDSO of Nu Amine for adult male albino rats
is 708p1/kg of body weight with confidence limits from uh7?1120im/kg.
Deaths occurred within 1 to it days, and toxic signs were mainly characterized
by depression, weight loss, labored respiration, ataxia, abnormal gait,
circling movements, depressed or absent reflexes, and hematuria.
A single application of 0.05 ml. of Nu Amine to the eyes
of albino rabbits produced marked irritation, including corneal.damaga
which did not completely subside within seven days. However, no evidence



of systemic toxicity from mucous membrane absorption of the test material
was observed.

The estimated acute dermal LDSO of undiluted Nu Amine for
albino rabbits is 1,2 ml/kg of body weight. A corrosive?type skin reaction

. fl-
was produced at dosage levels of 0.1, 0.5, and 2.0 ml/kg of 


In. 

At 2.0 ml/kg, toxic signs included depression, labored respiration;
depressed righting and placement reflexes, incoordination of hindquarters.

abnormal gait, prostration, sluggish pupillary response to light, absent

#4
C73

b3

as}

pain response in hind limbs, running movement of hind limbs, and coma.
Necropsies revealed definite liver and kidney changes at the 2.0 ml/kg

level and pcssible liver and/or kidney damage at the two lower levels.

Subacute Studies

Oral Administration Dogs

According to Drill and Hiratzka (2) oral ingestion of 2,h,5-T

by adult male and female mongrel dogs for 13 weeks at levels of 2.0, 5.0,

PTBLEQ M00

and 10 mg/kg of body weight produced no adverse effects. At a dosage
level of 20 mg/kg, total mortality occurred between days ll and 75, and
gross toxic signs included weight loss, weakness, slight hind leg stiff-

ness, difficulty in swallowing food, and bleeding from the gums.
Dietary Feeding - Rats

The results of a 90?day feeding study involving DoweJDl 97B

'(ester of was reported by Biochemical Research Laboratory of

. ?1
the Dow Chemical Company Ch).



Ingestion of Dowanol 973 by young male and female albino rats

for 90 days produced no adverse effects at dietary levels of 100 and 300 ppm.
At 1000 ppm, increased kidney weights were noted in the male animals and
histologic alterations were noted in the liversof both sexes and inithe

 
kidneys of_the females. At 3000 ppm, growth suppression and incrgaSed

1 


A



liver and kidney weights were noted only in the male rats, while gross
and microscopic changes in the livers and kidneys and increased terminal

serum alkaline-ph03phatase values were noted in both male and female animals.

18328





Oral Administration - Steers

Repeated Oral administration of Esceron Brush Killer (formulation

of ester of 2,h?D and steers, one per level, was also reported

by Rowe and Hymas (3). Results of these studies were as follows:

1. Fifteen (15) consecutive daily doses of Esteron Brush Killer at

a level of 100 mg/kg/day produced no gross toxic effects. How-

ever, necropsy performed h8 hours after the last dose revealed

some intestinal irritation and histologic alterations were noted
in the liver and kidneys.

2. Two (2) consecutive daily doses at 500 mg/kg/day produced slight
toxic signs an hours after the last dose but apparent recovery within

an additional 2h hours.

Three (3) consecutive daily doses at 500 mg/kg/day produced no

gross toxic signs.

Three (3) consecutive daily doses at 1000 mg/kg/day produced

death (three days after the last dose).

Consequently, assuming field usage at two quarts/acre, a steer
grazing on_7hh Sq. ft. of treated forage would consume a maximum of 100

mg/kg of the formulation which, as shown above, was not lethal?after 15

daily doses.

1.


Oral Administration - Sheep and Cattle

Palmer and Radeleff (7) reported the following results of re-

peated oral studies in sheep and cattle (oneanhnal per level).

Blues moo

18328

C8

The propylene glycol butyl ether ester of 2,h,5?T was lethal to
sheep following 369 consecutive doses at a daily dosage level of 100 mg/kg
of body weight and to sheep and cattle following seven consecutive doses
at a daily dosage level of 250 mg/kg. The triethylamine salt of 

produced no toxic effects in one sheep after #81 daily doses at a dosage

level of 100 mg/kg of body weight.
Repeated Dermal Applications Rabbits

The results of repeated dermal applications of Nu Amine was
also contained in the report to Diamond Alkali Company (5i, and a
supplemental report (5a) dated December 16, l965 presented findings from
microscopic examination of bone marrow from three animals.

Aqueous emulsions of Nu Amine - at field strength (h oz/gal.
or and at three (12 oz/gal. or and five (20 oz/gal. or 15.6%
times field strength were applied at a constant daily dose of 1.0 ml/kg/
animal five days per week for 20 applications to intact abdominal skin.
Moderate dermal irritation and evidence of systemic toxicity from
percutanious absorption of the test material were produced at all con~
gcentrations. Toxicity was characterized by depression, labored respiration,
sprawling of the limbs, weakness, and body weight loss; prostration,
incoordinstion or partial loss of use of hind limbs, depressed reflexes,

also noted in one animal at each

clonic convulsions, and death was



concentration. 


.
a

'Depressed hemoglobin values, increased_percentage of segmentEd

neutrophils, and increased urinary bilirubin values were noted in each

test group. Histologically, evidence of compound effect was noted in

STELES Moo

C9

liver, kidney, and skin at field strength and at five times field strength

(middle level tissues not examined).

No distinctive bone.marrow effect

was noted at three and five field strength.

various derivatives in fish and other marine species have been performed
by Ehe Bureau of Commercial Fisheries and reported in Fish and Wildlife

Service Circular No. 167, No. 199, and No. 226.

Fish and Marine Life Toxicity

Toxicity studies on 2,h,S?trichlorophenoxyacetic acid and its

AISLES MOO

Data pertaining to 2,h,5?T

has been excerpted and is summarized below.

Oysters -

0.1h of the butyl ether ester of 2,h,5-T caused
50% decrease in oyster shell growth (96- hour E050) with re-
covery within one week 2.0 of the acid of 2,h,5-T

produced no toxic effect (9).

Brown Shrimp -

Fish -

1.0 of the butyl ether ester of produced
10% mortality after 24-hours exposure and 20% mortality after
h8-hours exposure, and 1.0 of the acid of 2,h,5?T produced

no effect after h8-hours eXposure (10).

50.0 of the acid of 2,h,5-T produced no effect in Longnose
Killifish after h8?hours exposure 1.0 of the poly?lypol

butyi ether ester of produced 0.32% mortality in-Spots

after 2h- and MB?hour exposure (10).

Fri
CE



Ihe potency ratio of derivatives for Bluegill relative
to that of DDT was quoted in Table 2 (11) as 0.10.

I Although no specific studies or mortality data on only 2,h,5-T
for wildlife were found in the above Fish and Wildlife Service Circulars,
the potency ratio of 2,h,5-T derivatives for Bobwhite Quail, RingeNecked
Pheasants, and Mallard Ducks relative to that of DDT was quoted in Table

2 (ll) as approximately 0.50.



STELEQMOO.

pm;

CE

CA9





Conclusion

Although considerably more toxicity data, particularly regard-
ing subacute toxicity is available on 2,h-D than on 2,h,5-T, the aVailahle
toxicity data indicates that the two materials are comparable. The acute
oral toxicity of 2,h-D and 2,h,5-T and their various derivatives for labora?
tory animals is in the same range, 300?1000 mg/kg of body weight. Short-term
(90?day) dietary feeding of esters of 2,h,5?T to rats at 100 and 300 
level, and short-term (l3-weeks) oral administration of the acid of 2,h,5?T

to dogs at level of 2.0, 5.0, and 10 mg/kg of body weight produced no

SIBLES MOO.

adverse effects. Sheep tolerated h81 doses of the salts of both 2,h?D
at a dosage level of 100 mg/kg of body weight, while the ester
of 2,h,5eT was lethal to both sheep and cattle at levels of 2,h-D which

were only toxic or non-toxic. However, 369 doses of the ester

of at a level of 100 mg/kg of body weight were administered to one
sheep before death occurred. The toxicity of 2,h-D and 2,h,5?T for fish

and wildlife appears to be comparable. Particularly, since the long history

. of usefulness has not produced evidence of unusual toxicity.



C12

BIBLIOGRAPHY

Rowe, V. K., McCollister. D. D., and Spencer, H. C., The Acute
Oral Toxicity of 2,h,S-Trichlorophenoxyacetic Acid to Rats,
Mice, Guinea Pigs, and Chicks; Biochemical Research Laboratory,
The Dow Chemical Company, Midland, Michigan, 1950.

Drill, V. A. and Hiratzka, T., Toxicity of 2,h-Dichlorophenoxyacetic
Acid and 2,h,S?Trichlorophenoxyacetic Acid, A Report on Their
Acute and Chronic Toxicity in Dogs; Industrial Hygiene and
Occupational Medicine, 7:61-67, 1953.

Rowe, V. K. and Hymas, T. A., Summary of Toxicological Information
on 2,h-D and Type Herbicides and an Evaluation of the
Hazards to Livestock Associated with Their Use; American
Journal Vet. Res. 15: 622-629, 195A.

Results of Toxicological Tests on 2,h,S-Trichlor0phenoxyacetic Acid,
Reported by Biochemical Research Department, The Dow Chemical
Company, Midland, Michigan, August, 1950.

Toxicity Tests on Nu Amine (l,3-propylene diamine esters of
2,h?D and 2,h,5?T; Report to Diamond Alkali Company dated
February, 1962 by Hazleton Laboratories, Inc.

. Microscopic examination of bone marrow sections from the above

repeated dermal study, Report to Diamond Alkali Company, dated
December 16, 1965 from Hazleton Laboratories, Inc.

Results of 90-day Dietary Feeding Studies of Dowanol 97B (Ester of
2,h,5?T) in Rats, Report by Biochemical Research Laboratory,
The Dow Chemical Company, dated November 27, 1961.

Palmer, J. S. and Radeleff, R. D., The Toxicoloaic Effects of Certain

Fungicides and Herbicides on Sheep and Cattle, Annals c1
N. Y. Acad. of Sc. Bol. 729?732, 196A.

Butler, P. Commercial Fishery Investigations, Acute and Chronic
Toxicity Studies, U. S. Department of the Interior, Fish and
Wildlife Service Circular 199, 5?6 and 26, l96h.

r? .-

Butler, P. Commercial Fishery Investigations, Laboratory St dies
and Toxicology, U. S. Department of the Interior, 



Wildlife Service Circular 167, 11-15, 22 and 2M, 1963. .

0881.89 MOO




am

C13

10. Butler, P. Commercial Fishery Investigations, Acute Toxicity

(Biological Laboratory, Gulf Breeze, Florida), U. S. Department
of the Interior, Fish and Wildlife Service Circular 226, 69?73,

"l965.
11. George, J. L., Recommendations for Minimizing Dangers of Pest Control

and Pesticides to Fish and Wildlife, U. S. Department of the
Interior, Fish and Wildlife Circular 167, 101?105, 1963.

IZELES MOO

18335

INDIANAPOLIS . INDIANA

daft. --

WINTHHOP 2500


Dr. Harold H. Gay
Medical Director,
The Dow Chemical Co.
Midland, Mich.

5 I

FM 

1 ROBERT E. JENKINS. M. D.
1 3.1m N. MERIDIAN ST.

June 7, 1951;

Dear Dr. Gay:

Mr. James R. Mayfield has recently brought
in several letters from you and other inter-
ested parties regarding an alleged contact
dermatitis he may have sustained in August
1953 from the use of T.-or something.

tessta rum?

I have seen Mr. Mayfield on several different
occasions since his first visit here on Aug- 
net the 114th 1953. At that time he presented
a severely excoriated dermatitis of the peri-
anal area. From the history at that time he
was stated to have had pruritis ani for ten
years. There was some dermatitis of the el-
bow extensor then but I was not clinically im-
pressed with the dermatitis venenata picture.

He has been treated with various topical med?
icaments and with some success but still has
some dermatitis. I also had him seen by Dr.
R. Kierland at the Mayo Clinic and they felt
much as I do, namely that there might have
been an element of contact early but that
would not explain the pruritis ani of long
duration except from the "over-treatment"

 
  

  

standpoint.
SincereI I
.6:
Re ert Jenkins, MD

 


IO




CHEMICAL COMPANY

MIDLAND MICHIGAN

April 12) 195? .

Dr. J. S. V'sndenerk

Assistant Professor of Horticulture

Purdue miversity Agricultural htensim Berries
Department of Horticulture

Lafayette, Indiana

Dear Dr. Vsndenerkt

Your letterto our-Ir. v. K. Role isnow for
oment.

As Hr. Howe hes told you, this is certainly not a reaction
typical of contacts. I am assuming that my-
field has had no subsequent contacts with since the
original back in August, and if this is two then it
follows spriori that the demtosis which he now has, is
not the molt of contact with becsuse contact
dermatoees by definition are cleared certainly within two
nonths after discontinuance of the contact.

I: contact has continued however since that tin, then the
first 
further contests, aid it nest be remembered contact
does not necessarily neon direct contact with the material.
It my be-?contsot through inc-wearing or clothing uhich are
impregnated with the substance.

I hesitate to offer any suggestions for therapy without
knowing more than the fact that ?this area turned red in

the field and now remains rather red and irritated." I think
the most effective way for me to offer any help would be

to talk to some physician who has recently seen the skin
condition. If you could give me the name or the physician
who last sew him, I should be glad to cell and talk with him.

Very sincerely,

Harold H. Gay, 11.1).
Medical Director



CC: V. K. Rowe, Biochem. Res Dept., 280 Bldg.

 THE



MIDLAND MICHIGAN

June 5. 1953.

V. K. Rove
BIO-CHEMICAL LAB.
2?280 Bldg.

 

cc:
H. L. Smith

SUBJECT: REPORT OF CALL BY CLOFT, BOSTON OFFICE ON

LUCAS TREE EXPERT COMPANY

Attached you will find a copy of a report of call from
Harry Cloft wherein he requests some information describing
the effect of a 4% solution of 2,h,5~T in kerosene on

eyes.

Perhaps some action has already been taken on this matter;
however, Hillard Smith will be in town on Monday, June 8

Could you call Hillard Monday to decide what action,if_
any, should be taken at this time?

XJ.

Yours very 

a.~J
Clyde . Bryant 
Agricu tural Chemi a1 Sales
Ext 8804



1 2 4i '7

9898.431" 

16338

4



I PURDUE UNIVERSITY
Agricultural Extension Service
Lafayette, Indiana 

Department of Horticulture Egg
5..

March 24, 195k 
K:
Cb

Mr. V. K. Rowe :5

Dow Chemical Research Department to
no

Dow Chemical Company
Midland, Michigan

Dear Mr. Rowe:

I an inquiring as to whether you or anyone in your or-
ganization might know of a treatment for an apparent allergy
developed by 2&5 T.

One of our growers in this state received severe toxi-
cological reaction appearing to be a burn over a portion of his
body while spraying fence rows with 245 T. This occured last
August. This area turned red in the field and now remains rather
red and irritated. Ir. Hayfield was quite concerned over this
and local physicians were not able to give him any satisfaction
fand he went up to layo Clinic last January. They didn't seem
'to know any treatment for this type of skin injury. Could you
possibly suggest what might be done, if anything, or refer us
to anyone who is more familiar with this type of a toxicological

problem.

Very truly yours,

Joe

J. S. Vandemark
Assistant Professor in
Horticulture



HQO

main 0. 

OOHJW

whmu. 3.U.
0% ?In mwuu:
waDHHummH wwum.



9.5m m. 

92991315100

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nonm?nwm momma. 
mmu nuduu 

mm" hunumun 
4m. musmw onuwmua

. ?o UH won awn wmnnu uwmum mm won



Hmmumma w.

85m Hmm?hum w?m mm m: rm 

Odumm owwulnlullnunnunlur awn? ww?m

Madman mvwasunuulauunaluunmmaw<m 
mmqunlunanuaunu?L awn 

damwm ionwm Munun um ?aw nm?mm ow Hmnmun ?buu?mm mm-. 
- wan nusnm oww.nmmw mm dauunwm.wmd ?um dwdu mrm.?mmu wuwwmu.

I .L. .. ..-. 1 .. .. 
utwnh L: Hm orlu Ln?. do ?w chin Emu amorlmH nurwh m: ciwu 
.1by?. mu awn vam.- armn on ho: .c quIn ?mm

wuwvuwuu.
.I.T. maibldIrI  IIHJ.J flle.l (.0 Pals.an

OHthluunn  

mwunmumww woch~
w?mmcm b. 

h. mmsa R. U.



 . . 125s

conmanuv?

MIDLAND MICHIGAN

July 12, 1955

Dr. Andrew G. Goesl
Daniel-Harrell Clinic
Medical Arts Building
317 State Line Avenue
Texarkana, Arkansas-Texas

I Mm

Dear Dr. 60681:

Mr. R. A. of our Agricultural Chemical Division at
St. Louis has asked me to reply to your letter or June 25th
addressed to that office. 

I an very sorry to hear-that you have a patient-who is suf-
fering from mild hepatitis following use of our Esteron 2&5.
Our studies of the toxicology of this material have been con-
ducted on laboratory animals and livestock, and while mild
hepatic have occurred, the dosages in all cases have
been rather massive and by the oral route. Furthermore, ef-
fects upon the hepatic or renal systems seemed to appear only
after rather large doses were given and after other-more dras-
tic effects suoh as nuotonia,'snorexis, and gastritis were ex-
perienced. Thus, if the patient had swallowed the material

in substantial locusts, it lould not be unreasonable to-ex~
pect some transient hepatic and perhaps kidney inJury to follow
other primary 

It is recognized, of course, that such experimental work is
suggestive but not cone usive as far as human subjects are
concerned.

As you are probably aware, we have manufactured and sold Esteron
2&5 and related materials for several years. Millions of
gallons have been used and many people have been exposed. To
our knowledge, there has never been a case of systemic toxicity
attributed to these materials except perhaps where it has been
swallowed accidentally or with suiCidal intent. An occasional?
case of dermatitis has been reported,but in most of these cases,
the diluent oil (fuel oil or kerosene) has been shown to be

the causative agent. 

The significance of this plant and field experience is not ap-

preciated until one realizes that there are many custom spray-
ing firms in this country and abroad whose employees do little

18341

Dr. Andrew a. Goes]. 2 July 12. 1955

else but apply these materials. It is not unusual for these
men to have their clothing wet with the material for hours a
day, day after day. He, or course, do not recommend such prac-
tice and do our best to discourage it, but nevertheless, it
does occur. 

In view of our laboratory findings and the experience we have
had, we doubt very much if the application or Esteron 245 by
your patient could have been in any way related to his hepati

tis. 

I am sending a copy or your letter together with a copy or this
reply to Dr. E. H. Gay, Director of our.Hedical Department, and
thereby will ask him tc-contact you directly with his opinions
in the matter. 

If we can be of any further assistance, please do not hesitate
to contact us.

Sincerely yours,



V. K. Rowe

2-280 Building


enc.
P.S. I am enclosing a reprint entitled, "Summary of Toxicologi-
cal Information on and Type Herbicides and
Evaluation of Their Hazards to Livestock Associated with
_Their Use," Recognizing that this information does not

apply directly to human beings, we believe it is indica-
tive of the type of thing which can be expected.

V.K.Crandall
H. L. Smith
G. J. Williams

1193243 1 M00:




33.83



2

 OF TOXICOLOGICAL TESTS

ACID

U)

Biochemical Research Department
The Dow Chemnzal Company
Midland, Michigan
August, 1950'

PROBLEM

The widespread handling of 2,4,S-tricnlorophenoxyacetic a 10


:lde it desirable for us to have a significant amount of toxico
logical information.. Data on the oral toxicity was necessary for

presentation-to the F.D.A. Hearing on residue tolerances.

MATERIAL
Name: 2,4,S?Tricnlorophenoxyacetic acid
Formula: C1
Structural: Cl
Cl
Empinical: 08H501303

EXPERIMENTAL RESULTS

1259

81.8899 M00

The acute oral toxicity of 2,4,S-trichlorophenoxyacetic acid

113C.

for rats, mice, guinea pigs and chicks is given in the attac?
summary which was presented in evidence at the F.D.A. Hearing.
.concluding our testimOny on this material we stated,


3
Q3

CE

5?

 ?by 

0f the SiMilarity in chemical structure and in
toxicity and physiological effect between and it'
seems that a residue tolerance for the latter on fresh fruits and
vegetablcc.should be the same as that established for 2,4?0; it
would seem that a residue of 20 p.p.m. would provide a wide margin
of safety".

Irritation

When 2,4,5?trichlor0phenoxyacetic acid as a 10% solution in

81.8899 MOO

propylene glycol was introduced into the of a rabbit, it

produced marked pain and slight conjunctival irritation immediately.
Five hours later, severe conjunctivitis and moderate'corneal damage
was apparent. This picture remained about the same for two days
before healing began. Five days after the exposure mild conjunc-

tivitis and corneal damage was still apparent.

I
I

kin Irritation

 

when 2,4,5?trichlor0phenoxyacetic acid as a 10% solution in
butyl_carbitol acetate was applied repeatedly to the rabbit ear and
bandaged repeatedly to the shaven abdomen,-a slight simple irri-
tation developed in both instances. I

SUMMARY

 

The toxicological preperties of 2,4,5?trichlorophenoxyacetic
acid may be summarized as follows: I

1. The material.is moderate in acute oral toxicity for the
or these
species the rat seems to be the more resistant and the
chick the most susceptible.

four species, rat, mouse, cavie, and chick.

2. The material is markedly irritating to the eyes.

3. The material is to moderately ir itating to the
skin upon prolonged exposure.



xi

yer.?
CE
us


 I

all

amallowing

HANDLING HAZARDS AND PRECAUTIONS 303 SAFE HANDLING

 

It does not.seem likely that 2,4,Sutrichlorophenoxyacctic
acid Can be swallowed accidentally in amounts dangerous to life,
although this might be possible if concentrated solutions of the
iatorial were available to children.



0?



If substantial quantities
the material'were to be swallowed by anyone, serious ill effects
could be anticipated.

If the_material should be swallowed, induce vomiting by giving

1 common emmetic such as 2 tablespoonfuls of table salt in a glass
3f warm water. Call a doctor.

?18899 'Moa

 

The 2,4,Sutrichlorophenoxyacetic acid is capable of causing
serious damage to the eyes. Precautions should be taken when
.andling this material to prevent possible contacts. It is

ommended when handling strong solutions of 2,4,5-trichlor0phenqu?
cetic acid that goggles, face shield, or other similar device he
orn to prevent accidental splashes in the eye. It is believed
hat the handling of the solid does not present a hazard simply
ecause of its insolubility in aqueous medium.

If the material in any form should get into the eyes. they
'ruld be washed with flowing water for at least 15
inutes and medical attention obtained.

(in



Prolonged and repeated contact with the skin may result in

'ritation. Occasional contacts for short periods are not expected

a cause irritation. Strong solutions are no doubt more likely to

.use difficulty than the dry solid;

In case of contact wash the exposed area with soap and water.

not wear clothing or shoes which have been contaminated with solu-
ons_of the material. 

lags





Inhalation

 

30 studies of toxicity upon inhalation of 2,4,5-trichloro?
phonoxyacetic acid dust have been made. We believe, however, that
in view of the similarity with this material to that of 2,4-dichloro-
phenoxyacctic acid, that it is unlikely that toxic ooncentrations
3f the dust wou1d_be inhaled, pri?arily because of the irritation
1t_wi11 produce upon the upper respiratory passages.

SL8899 MOO

18348



?cLon; 

?ti-r



TOXICITY OF ACID AND
ACID

A Report on Their Acute and Chronic Toxicity in Dogs

VICTOR A. DRILL. MD.
AND

TOMIHARU HIRATZKA, MD.
DETROIT

BOTH 2.4?dichlorophenoxyacetic acid (BA-D) and 2.4.5-trichlorophi-nostacetic
acid may act as plant hormones and herbicides' and are now being
fairly widely used {or the control of certain types of weeds on farm leads. The}
are being sprayed from airplanes on many acres of Specialized farms and grazing
land and are present it'. trademark preparations sold a: retail for home gardener
\Vith the widespread use of these herbicides. possible toxic e?'ccrs are important.
as accidental ingestion may occur or residues may be inadvertently encountered in
food. In an earlier report. 2.4-0 was mentioned as being nontoxic to animals
and man when administered orally.??1 However, the injection oi large doses of
in animals produced S)'Illp10m5 similar to those seen in clinical nit'omnia.?
More detailed observations were reported by Hill and Carlisle who Studied the
acute and subacute effects of in various species of animals.? There hate been
no reports concerning the toxic properties of The present study concerns
the acute oral toxicity and chronic oral toxicity of 2.4-1) and in dogs.

EX PERI MENTAL PROCEDURES

Adult mongrel dogs of both sexes were used. Most of the animals were housed in the
laboratory for a period of two to three months before the study was started During this
control period they were immunized againsr distemper with the Green vaccane. The dogs were
fed a standard stock dict (Friskies) ad libitum. Both and 2.4.5-T were commercial
materials. with a purity of 93.5 and 98.9% and freezing points of 132.8 and 150.6 C.. respeCtiirelt-.

In the acute studies. the calculated dose of 2.4-D or was administered as a single
oral dose in capsules. The dogs were observed for a period of 14 days. at which time the sur~

 

This study was supported by a grant from the Dow Chemical Company

From the Department of Physiology and Pharmacology and the Department of Pathology.
Wayne University College of Medicine.

I. Hildebrand. E. Ms. War on Weeds. Science {03:465-468. 19-16. (15) van OverbeeL.
L. and V?lez, 1.: Use of Acid as a Selective Herbicide in the Tropics.
Science ?13:472-473. 19-16. Smith. F. C. and Carlson. R. 17.: Control at
Ragweed Pollen Production with 2.4-DichlorOphcnon-acetic Acid. Science 1113:473-474. lQ-l'i
(J) Martin P. C., and Mitchell. J. W.: 2,4-Dichlorophenoxyacetic Acid asja 
Herbicide, Bot. l9-l-Buchcr. N. L. R.: E?ects of 2.5-Dichlorophcnoxyacctic Acid 9n Emrim?nf?ar ?An'mls'

Proc. Soc. Exper. Biol. Med. 63:204-205. 19-16. I . -
3. Hill. E. V., and Carlisle. H.: Toxicity of 2.4-Dichloropltenoxyacettc. Acrd for Expert-
mcntal Animals. J. Indust. Hyg. 8r Toxicol. 29:85-95, 1947.

61

Pesos



Q22





63 LYDUSTRLU. Hl'lillfl'li 1H 

.viwrs \u-rc autupsictl. In the chronic studies. the or ?nu mimiuislcrul 
[ti capsule: fun- day: a wet-L- uvcr a 1.5 ?rel; pcriud. Each capeulc WM in a 4 to 3 pui
plet'uf of tanned dug laud. ?hid: the attituals cumulncd fC-Nili}. 11m: :uuiduu; the
continued trauma of the stunmch lube. During tlu: study the aninmh m-rc .t
?(ck Control blood Cuutlti ?are takrn bciun: tlu: arhninistratiun oi the (Hug, nu the 301? 
and on the 90th day at the of (In: study. Hcinugluhin dvtrilnmn'ii ?uh 
l'pou ?it:th or at the cmtullction oi the study. tlu' animals ?we:
lumpii?l. Tl-?r?ucs were takcn (mm the lung. heart. liver, kidney, adrenal". thyroid, aml

0" Rites. ?xed in 10% formalin. and stained with hemaluxylin and cmin.

TAILS l.??E?ect of .S?Engh- Oral Dom o] Acid and
Acid in Dog:

 

\"elu ht

 

 

 

. DOW: Death
Doc Hthg. Loss. Kg. Dry: Hort-lit2.?1ans-T 
a 3L. .. mo ?I.l 2 
31 .. ?t.i 
15 .. mo ?o.5 a 
.. mo --1too ~04. 
19 .. 50 +0.3 0 a 0.1
an+0.1 0 a
:1 .. 50 -o.c a 3

 

. 

In this column means lent-1:; male.
[Column Indicate: truer-l unruly of syiatploms din-113ml In text. a mum: no change from normal
83 la column mun: survived an tut period.

RESULTS

1. Arm: Oral Toxicity?(0) Mortality: The effect of single oral do?: of 2,4 l)
or on survival for 14 days is shown in Table l. The deaths. were 
and occurred two to mi days alter the Compounds were administered. The oral
of ?as approximately 100 mg. per kilogram of body weight; for
it was in the range of 100 mg./Lg. or higher. 

Bod}: ?'cight: The large oral dnses of 2.471) or: pruducrd a
decrease in bad)- weight (Table 1). Such animals- dc'tflopgd Furious dc'grct-s of
anorexia, and those that {lied refugd even canned d'og 't'o'od toward the end of their
survival. 

 

4. Emlyn. K. A.: A Stabilized with Light J. Biol.
Chem. ?5:63-75, 1936.

 ?hiv?i'i'K'Hilfx 

In dogs that died the etleet prudnt?ed In 2,4-l) hunt a
mild ataxia and stiffness in the hind legs to a definite nnutnnia. Initial signs were
often noted six hours after the oral atlutinisttatiun of this time the animal-
were more quiet than annual. and a slight ataxia. was occasionally garment. The hind
legs were always affected first; the lnrelegs later not at all. 'l?ht-ri- 

a progressive increase in spasm in the hind legs with inert-amt; ill??llt? 
and some-drags extended the hind lintl-s in a spastic mm mnent Inning:
a few seconds. The kin-t: jerk was either normal or hyperactive. 
in the early stages of acute poisoning witlt the aniutal~ nt-ri- n~uallvs
quiet. [i the animal was lifted and made to walk. lhe spasm and ataxia \u?tr L'lt'illi}
noticed. but after it had walked for a while, the spasm and ataxia tenth-d tu ?ll?i'ftn?l?m
In the later stages of intnsicatiun. the animal. when placed on its iCt?l. wa~ umhlt- it)
stand and would then sit or he in an awkward position with spastic marker
particularly by spasm ?if the hind legs. Other dogs anti rulled Hit the tint-rm
in attempts to right themselves. occasional dug slim-red evidence ni irritatinn ?arm
pain when the skin over the hack of the neck was grasped. Home dings. \thu'n 
up hy the shoulders. would show extension and crussing oi the hind lnul-- 
toxic effects of the drug progressed. the (logs eventually refuse-d the Hull-lb til-w
of food oiiered. Occasionally, sneezing, ruhhing oi the and diarrhra wen
observed, hut not vomiting.

Such signs were also seen in one dog that survived a lower dnst- ni int
14 days (Table in this instance the changes were relatively mild and were
limited to spasm of the hind legs and a slight Ataxia.

Marked effects were observed in the (lugs receiving 'lhr vhaugi -
were limited to a slight or moderate stillness in the hind legs. with the tlt'H?lHl'HW??
of ataxia in the two dogs receiving the highest doses, N0 Hitlt?ltl
in the one dog that died after a single dose ?ii 100 mg [kg ('l'ahlc- 

Pathology: Changes ohserved were limited to the trail Jill-l
lungs. Four dogs 2, 4, and 6) receiving 2.4-0 and two (lugs 1.3 and 17; ml
showed mild to diffuse redness of the mucosa in the small lu annu-
dogs pneumonia was present. llistnlogicallv. the nitrate tings showed snnu- llt'l?flt?l-
and in?ammation of the intestinal mucosa. One dog (1.3) had a ninth-rate. tliti?u..-
hepatic necrosis. Two dogs (l3 and 16) had a mild degree of renal tulntlar tit-gm
eration. In most dogs that died the ?ndings were limited to clianet 
such as hepatic congestion. in many cases death was apparently due to 
which followed the development of anorexia, weight loss. and 

II. Citrmiir Oral Mortality". .\ll dogs receiving 3. S. or 10 in;
of 2.4-0 or 2.4.5-T per kilogram nt' hudy weight survived the 90-day periu-l
The dogs receiving 20 tug/kg. of these cuinpounds died during the stud}. Tha-
administration of 20 rug/kg. itt divided doses twice a day produced death in hoth
dogs receiving and in one dog receiving 2.44) (Table 3).

Body \l'cight: The only signi?cant changi- in. lintly_\_veigltt occurred in the
animals that_lailed to survive the study (Tahle 2). Loss of lit-eight in till?? dogs

began 7 to 12 days before the death of the dog. - 
General Dogs?that survived the?nrai tit minisfratiun oi 2.4-D nr
for 90 days were free of any  

1834195

64 

The three that dit'd \vhile waiving the highmt tluw ui 3.4-1) 
differing somewhat innit tltosc- ulrervt'tl in the mute studies. The tluga \vt'n- mi:- 1.
tveaLer, and less than normal. ?as higher in lltt' him! it
particularly on passive exteusimt. 'l'he hind legs tvt-re held mure than 11 -5
when walking. and :1 slight ataxia was present. During the tun [11 thru- 
of their survival tuu m' the dogs shmved dif?culty in chi-um}; nr uvulltmiug I
eventually even refused a small bolus oi canned dug loot] (usuztlh ti"-
sumed). There was also some oozing of blood from the gums and 

TABLE fir?Body lVright' and Surriz-ul (If Dog: Fm? flu}! tutJ
Arid [Hr 91) days

 

 

Weight.
Dose.  
Do: 111.- 112.1142. I"ii?" ?33? 
1.1.11
23? .. None 11.! 11H 42? . 
21 .. None 915.2 131211.1 11.11 +01 5 C71
1011 .. 10 1111 1111.2 11.11 -t 6 
:2 11 . 101 11.0 1311:11 .. 20 12': 10,1 111 
15 11 .. :0 13None 1.3 +01 5
8711 . . . . . . . . . . . . . . . . . . . . . . .. None I'12119 10.0 4 1.1 
?1 .. a 111211.1 11.5 5
1.1 .. 10 13.0 12.: 3 
1? .. 10 9.11 9.2 ?06 
15 11 .. :01 11.3 9.2 w? 5 49
.. :01 10.5 1.9 --1 a 1:1
111? .. an 119 12111this column mum lemalr; male.
9 Dilly dose d1?. hie-l amt. om- hatt In 11 111. end one-hull in p. m.
:5 In this column mum :univeut U1 any tut. period.

In dogs that died while receiving the prominent effects ut't'c ?cult-urn.

slight stiffness in the hind legs, dif?culty in swallowing food, and, in (me 
bleeding front the gums.

Blood Count: The administration of or 2,4,5-1? (lltl nut ltme an}
signi?cant effect on the hemoglobin, red cell enuut, or total white cell nt
animals that survived or died during the study. The differential cumtl renmim-tl
?normal in the surviving animals. In three animals-that tli?d a terminal fall in 
percentageh'oi was ohserved (Table 

(6) Organ ?Eights: Thergtvas no the ?eight ut? tln'
thyroid gland, adrenal gland, heart, liver, or kidney iu-lanimals that 1hr
90~da_v period of study. In two of the three dogs that died during the 

#5431




44"

70

00w 668861

IN-IRIHCIUI: 5' (-5

of there was a slight increase in and kidney weight. :nnilar clung--
was noted in two of the four dogs that died as a result of 

Gross Pathology: Two dogs receiving (26 and 34) are.? of
redness in the duodenum. With the administration of tln'le (ll'Yt'lUlK?fl in
deg 42 a diffusely reddened duodenum and jejunum. while in dog 43, ml
the 59th day, a generalized icterus was demonstrated.

1?55!? 0f 2.4-0 dud 3.4.557. cm "euloglohin, Tom! BloodDi?ereotinl Count

J- -. i.

 

Eh. 3110. Kent", Moo . Bh- . Emu; .
Doc No.? Du Gm. '13 Ill. ?'80 '5 1?3 ?In 

Control Don
None 0 13.20 572 11,300 5.93 15.603 64 :9 0 1
to [3.10 09 ?.350 1! 2? 2 0
14 l? None 0 6 0  10.? 86 II I 0
an 165 13,l50 7'9 1'2 1 I
I) ?.33 6.16 1.100 79 1] a 
. . None 0 0 60 6.03 16,109 26 0 5
I) 10.75 5.63 13,300 511 1r. 2 I
on 12.7! 5 ".101 Rune 12 19 6.53 14.500 5.3 2 I
I) 13 3  5.18 H.909 53 26 0 1
13.95 6.47 3.1013 um 13,500 61 21 5 <1 1
1.1.11 10 15,100 12,100 211 14.11.9,100 I (I 1
49 11.35 5.67 10.500 :15.2110 ?.95 647 13.150 76 II I 
2.5 11.59 on 9.100 68 27 I 1
2.4.51"

?5 I . ..  0 1151 5.15 1.6.100 911 16.158 8.01 6.1.10 11050 63 29 2 0 
11 6.82 H.200 'n 0 1
.. so 0 1?11 5.13 11,301 12.13 a tn 10,1100 51 2 0 3
I) A 11,550 7.175 In 9 I 

 

'1 In lhle column rut-emu hmnle: 51, male.

1953'] do" nod one hall to e. on. em: one hell In y. In.
To ?weenie apnea. only due on control don and the htgbeu (1021 are liven.

II.
I

(9) Microscopic Changes: '1 he heart, lungs, Ill) roid, adrenal, mar), te~tr<

did not demonstrate any signi?cant changes. Three of the. (Logs rewivmg 'l
and two receiving an occasional area?o'f' {pcal?inegrmis in the Inc:
This ?nding, however, has not ru-tated to either the, dose of the? cumpuumlu or the
death of any animal ("fable 4) and was judged tn be of no 'l'lu- dun
denum of dogs 26, .34, 42, and 43 showed valuing degrees of hyperemia and wim-
early in?ltratiOn of cells in the mucou. The snhinucosa and serosa were 

1mg

1 :5

C9





DOW 688862

.66 

A slight increase in the of casts in sections ?at Ihilt?ti in .t 
(Table but again this was unrelated to dosage and was of tluuhtlul 

COMMENT

In the acute studies, single large oral doses of administered lu \itlgn n. 
Pmd?Ce Sigus characteristic of clinical niyotnnia. This effect of BA it run Hill-Ll
to that observed after various experimental species had received In,
addition. some dogs showed signs of irritation and pain when the skin hi tltt' itJt'iu 
?hc "la-"k ?'35 grasped, which may indicate meningeal irritation. 'l'lu- 
crossmg of the hind limbs, when lifted by the shoulders, may point to a spin?: Ht

Tm? in Um and Kidncy 0! Dog: Fed 2.4.0 or 2.45.1 m:  

Liver
. I
Defy. Death Foe-I 
Do: No. Make. Du Congestion Necrosis (?nts Tutu. 
men
23 .. None . 0 0 0 0
.. Kane None Kane 'Abo a 1+ ?brosls.
Thane: untied on but: of 0, and 

central lesion. Other e?'ects occasionally observed were sneezing, oi m-
eyes, and diarrhea. The effects of previorisly studied, it ere not as nt"-I Ir
being limited to stiffness in the hind legs and ataxia. The higher dost-s of mi - -
compound produced anorexia and weight loss. Although initial effects may he  
six hours after the compounds are given, the developed slant) in [its
dogs, and death was delayed. occurring several days afterr?t?he administration m' 
compounds. t- 

In the chronic studies, the dogs survived iS?o?'lO mg. of 
per kilogram of body weight without apparent?illi'e?ects. Hnu't't't?r, 
daily oral administration of 20 trig/kg. of 2.4-0 produced death in three of (mu
dogs within 18 to 49 days. This delayed death may indicate a cumulative 
The signs observed in animals dying from the 20 mg./ltg. dose of 2,4 ll



DOW668863

TZKA- TOXICITY Ul- 67

differed somewhat from those seen in the acute studies. With the clnunic mhninit
tration, the animals exhibited chie?y a stillness ot' the hind legs and weak
ness. dif?culty in chewing or swallowing. and occasionally bleeding from the gum.
All lour dogs which received 20 tug/kg. of sucCunthed between the 
and the 75th day of the study. Toxic ell'ects in these animals consistently 
slight muscle spasm and dif?culty in swallowing food. Thus. the urnl 
dose for in dogs seems to be in the same range as that obtained with 2.4 ll
With both drugs, there was a terminal loss in body weight.

Dogs surviving the chronic doses of or 2.4.5-T did not show any signiti- -

cant change in hemoglobin, total blood counts, or differential blood count. in thru-
ol the dogs (64, 57, and 105) which died while receiving 20 nag/kg. of nr
there was a de?nite decrease in the percentage of in lln'
peripheral blood. Hill and Carlisle have observed a fall in the 
leucocyre count in one dog and a decrease in the per cent of in tun
dogs to which 50 tug/kg. of 2.4-1) was administered intravenoust each day lur six
days. in mice which received chronic injections there was no alteration in the
peripheral blood picture.1

Large doses of administered intravenously or, in the present study. orally
occasionally produced some necrosis in the liver. However, the ni
chronic oral doses produced only a slight and inconsistent focal necrosis of donhtinl
signi?cance (Table 4). Deaths during the chronic oral administration of or
were not correlated with signi?cant lesions in the liver, kidney, or other
organs examined.

suststanv

The acute oral of 2.4-dichlorophenoxyacetic acid (ll-D) in 
approximately IOO mg. per kilogram of body weight. Doses in this range or higher
produced a de?nite myotonia accompanied by anorexia and weight loss. The acute
oral for acid was in the range of 100
mg. per kilogram or higher. Such toxic doses produced only signs of a mild
spasticity.

All dogs survived the oral feeding of 2, 5, c: 10 mg. per kilogram of or
5 days a week for a period of 90 days. These doses did not produce any
or changes in body weight, organ Weights, or blood count.

Three of the four dogs receiving repeated doses of 20 mg. per kilogram of 
died. and all four dogs receiving 20 mg. per kilogram of 2.4.5-T died during llac?
study. Toxicity to at the dosage level was accompanied by bleeding from the-
gums, necrotic changes in the buccal mucosa, and some diiculty in chewing and
swallowing, but there was only little evidence of clinical myotoriia. A terminal lull
in the percentage of was observed in three of the animals.

Death during the repeated administration ol 2.4-9 or was not related
to pathological changes in the liver, kidney, or other organs examined.

P35.
C73
w-

THE ACUTE ORAL TOXICITY 0? 2,155?
ACID TQ RATS,
mm CHICKS

K.  
D. D. MCCollister
and
C. Spence:

Bidchemical Resea?ch Laboratory
.'The Dow Chemical Company

Midland, Kic?igzn
11950 I

1261



1835

-1-

C-JLL

Single oral doses 0: acid
- percent suspension in olive oil by stom;cn tuLa

to se? groups of rats, mice and guinea pigs. Aliquots of a 3

laboratory.' White nice were obtained from the Carnorth Ferns and law

Hampshire Red chicks were purchased from a commercial hatchery. Kat??e,
young adult animals were used in each instance except for the chicks
which were approximately three weeks of see.

Table I summarizes the and volumes of olive

assess/?00

all solutions containing 2,h,5-tr1chlorophenoxyacetie acid administered

to each species.

ISINGLE ORAL or
ACID

 

Soecies _Percent material yblume Given by Stomach Tube

in olive 011 Minimum Leximum 
K. I. 

 

Rats 10  2.9
.Xice . go I _o.o3 0.3 
Guinea P133 '10 '3 I 1.0 . 2-9
5:1ch 355 - 0.3 5.8

 

Mortalities resulting from the various single oral dosages
0? 2,4,5-tr1chlorophenoxyacetic acid to the experimental animals are
Sivan in Table II. All surviVing animals were observed until it was

certain that they had fully recovered (usually about two Weeks).'


?Tatons of poisoning, particularly with the higher doses, were



ataxia. and avotonic. 3   

-2-

TABLE II


KORTALITIES RESUDTING FROE THE ADKINISTRATIOX or 
ORAL DOSES 0? PHEHOKYACETIC ACID ?0 
PIGS AND CHICKS
Rats- Mice? - Guinea Pins Chiqgg

 

- No.fed ?b.d1ed No.fed fazdied ?de?d??onied ro.rca :o.d:The acute oral LDSO values with their 19/20 confidence limits
were determined for each_species by use of the method described by
Litchfield and Wilcoxonl. The results of the statistical treatment
of the dosage-response data are presented-1n Table 

TABLE 112'



ACCTE ORAL LDso VALUES FOR ACID was:

 

ADHINISTERED T0 RATS, MICE, GUINEA PICS AND CHICKS
I confidence limits) Slope a
Soecies . gm.[kem. 1 ?Function?
Rats . 0.500 (0.391 0.6t0) 1.h9
nice 0.389 (0.2h5 - 0.619) 1.80
Guinea Pigs 0.381 (0.307 0-472) 1?21
Chicks 0.310 (0.211 - 0.455) 1.u0

the fold change in dosage required to produce one unit
standard deviation in response along the line.

in;
Ch

,1
35



1. Litchfield, J. 9., Jr., and Wilcoxon, F.: J. Pharm. and Expt.
- Ther. 3g; 99, 19h9.

 

1 I. -..-
dung-Han.

The Libchficld and Uileoxon t-sts for patella of

?11res and estimate of relative potency revealed that the 2,h,3-tri?
chlorophenoxyacetic acid was significantly (19/20 probability) more

D.

the following comparisons: rats versus mice, rats versus guinea 
nice versus guinea pigs, mice versus.chicks and guinea pigs versus

chicks.

toxic to the_chicks than to the rats.

There were no Significant

A

result 0;



if?erenccs in either parallelism relative potency as 

i




to";

998899 M00

1835?

I . - 
 5-K 1 
1p,_ 
.

Bicclu mical 

The Don Camical 


as.



raresvlg?

RESUME 0F SKIN TEE-TS on: 
1. IN Dow's T17.4-12-9
(x 2777-uo) 2777-40
2. lrmunnms 2,u,5? 277741
mxcmoaomzmr. m1}: FROM arm-1:2
(E 277741) 2777.43
CONCEHIRATED nmnn-zs FROM 237,2- 12194
mom news 1,2,3, Rep; Park A u.
(K 277742) 7 - - 
IMPURITIES FROM Hooxm's
(K 277743)
muss FROM vows 
(K 2696-5)
1mm FROM mom?s menmno?
mm (1c 2696?6) 
mus REPORT 15 THE PROPERTY
OF
THE 00w cannon. Conn" 

I

3x
?3
?3




?9

 

- F?gm?




?rlill CIFIICHRIC3IKI. 
July 12. 1955

HIGHIMI

Mr. R. Reese
Bisphenol Plant
3N9 Building

SUBJECT: SKIN IRRITATION OF SOME IPFURITIES IN 2,3,5-
AND FROM
VARIOUS SOURCES

3818.99 Lh?i??

The skin tests on the impurities in 2,3,5-tri-
chlorOphenol made from various sources have been completed.
These tests were undertaken to help explain, if possible,
the differences in skin responses produced by various sam?
ples of Previous tests have shewn
that only the impurities round in Hacker's 
phenol were capable of causing skin responses which were
unusual. ther possible impurities were shown to be similar
in their effects on the skin to 2,4,5-trichlerophenel.

The materi 1: submitted for testing were as fol-

1. Concentrated impurities from Dow's 2,u,5-
triehlorophennl (K 

2. Concentrated impurities from 2,u,5-triehloro-
phenol cede from ?eoker'e l,2,3,h-tetrachlorobenzene
(K 2777?41).

3. Concentrated impurities from 2,h,5-tr1chloro?

phenol from Dow's 

 

   

I 

     
 
   
  
   
  
  
  
   
   
    
  
  
  
  
  
  
     
  

 

..
.yzimu;

Hr. R. Reese - July 12. 1956


I

h. Concentrated impuritiee_from Hacker's 2,3,5- 
trichlorophenol (K 2777-33).
All of these materials were applied to the intact
rtbbit?s skin as a 10% solution in Dowanol SOB. When
tested in this manner, all of those materials gave ocean?

tielly the same recponee in that they were irri-

eatesvl?HMQ

teting to the rabbit's ekin. Thiz response was character-
ized by slight reddening of the skin and slight exfoliattui
or scalineae. This is conzidored typical of the response -
caused by 2,3,5-triohloropheusl at this concentration.

These results show that the impurities, as oo-
teined in these earplea, should not contribute significantly
to the irritating properties of 2,4,5-trichlorophenol regard-
less or source of the raw material. This finding is not in
accord with those reported earlier in which it was suggested
that an inmurity of Hooxsr': may
canon eoneform dermatitie in humans (See letter or January
27, 1956).

Two poeoitle explanation: for this difference in
response mere suggested. Tie first wee that the "acneform'
effect was wank :n nz;ure :nd may have been overlooked in
this eerie" or tcazo alth:?gh they were repeated on several
rabbit: with no eignlfica.t change in response.

The recon; possible explanszion has that there may

have been differences in the :zginal samples used even

 

 









no

alg?


FER-uni

Hr. H. Reese - 3 - July 12, 1956

though the impurities were concentrated in a 31milar manner.

Consequently, the samples were analyzed by infrared scan-

ning with-tne following results.



ANALYSES OP 



2,4,5-tri-
chlorophenol 2,4,5-tri-
made from chlorophenol

Dow's 2,h,5? Hacker's made from_ Hacker's
Identification

trichloro- tetrachloro- Dow's tetra- 
phenol benzene chlorobenzene chloroohenol Cf Constituents

re
3 9 1 3 91 2,h,5-triehloro?
phenol

9 
phenol


i 2,5?diohlorephenol

i.

.5 7 i 1 1.

49~5
5

10 
- mothozxpehnol
(1rpur1ty A)


methoxyphenol
(impurity B)

The remaining :31271313 in the above samples are

in structure to tn: methoxy

not identifie: but are similar

dichloro phenolo and dirc?hrxy benzenec. This

would indicate pcsaib?lin; cf Hi-et?rxy chlorc phenol:

and .- - and ?esorninOI.

ainilnr analysis was no: 

on the previously "concentrated. from

Hooker'n 1: in wgaevvr,

g~vo - 
16381

that t?c oenple repartr? Cn hurt

 

 

Hr. R. Reese July 12, 1956

from materia? o; a high orVer of purity, being 91 i_2 5

of I: is reasonable to assume that
any impurities that were in the submitted sample were present
only in minor amount in the criginal product thus reducing

no

the possibility of its produc?rc the skin effects observed

QBIEQPLMUH

in the first sample.
The skin irritation tests :n the two samples of
irpurities from tetrachlorcbenzene have been completed

also. The samples submitted were:



14?"  

2696-5 Residue from Dow 1,2,4,5 tetrachloro-

up!


2696-6 Residue from Hooker 
chlorobenzcne.

When applied reportedly to the rabbit's skin,
beta of these materials: as 5% solgtion in Bowsnol SOB,
caused only very slight irritation and slight to moderate
exfoliation. A similar response was obtained when the
Hooker residue was applied as
a 15 salutton. This r::p:nse 1: t"pical of 2,h,5jtri?
a; this level. Hence the impurities, as
represented 3n ese two should not affect mata-
risily t1- rcupensu of 

information proves useful t: youHalo, plea:e let me know.

721,14; add/wt-

Her): A. Wolf 
Biocremicsl Research Durartmen:
le?cya Building,

 

1.9: u" . . 

.x a hhouahonun 

ww? .m I ucuam Hos-cauam



m3. LuucH gondunom Hananno nunaa

Wu, mnam zohdonmm Hanucoo shun-o:
anuucoo






hucsom .m noudonom no youOOhdn


QCONC
saga uoapauummH

Hoconnxuh.yun
oonuanoauu-m.4.~ aonu 

1.41.43;

. .. .aohu
.Hocuam "oadz .H.m.u

If
ll



.copnw and noouson

.. ?anaconunuauum.?.m.a unu Hanunnonuanownunm.w.m
.maumu%MIUhu Jwauwhao?a uaowuub no name? cwxu no 

BHKD

 

.ll'

.4 . 

. .31h-Lnn-l:

. 38L -  - 3 a 9
221?: THE Dow CHEMICAL COMPANY 1 2 0 

NEW YORK .
IAN .
PHILADILPI-HA MIDI-AND MICHIGAN 

Low ANGILII Ill. u. s. In. on.

 

Doceaber 9, 1949 
CLEVELAND a
 2' 
3:3: 5" 
soc-row vessey ?3
Iain Office c?
#7 Building

  

lubJect: Request for authorization? -
acute and cub-acute toxicity tests
on dogs with 2,l-D and 2,4,5-1.

(1) In preparing toxicological interaction suitable for
presentation at the IDA hoarince on agricultural choaicalw, wo be-
lieve it would be desirable to have inrorwation on tho acute and
sub-acute toxicity or and on dogs. This work will
be in addition to aiwilar work which wo have conducted on swaller
laboratory aniwals and in addition to that which appears in the
literaturo. rho advisability of conducting this work has been dis-
cussed with representatives of tho Agricultural sales Dopartwent and
tho Legal ,s

(2) Is have discussed this problel with Dr. v. 1. Drill,
er.the Depart-sat or Pharwccolcgy, School or ledicinw,?iayne
torsity, Detroit, and have received word fro: bin that he is

willing to undertake this work; It is our feeling that a fair price
for tho work we have in wind would be about three thousand dollars
(l3.000.00).

(3) we therctore requcet that an authorisation for three
thousand dollars be set up to cover the acute and sub-acute toxicity
studies on dogs with and 

you have any quostions regarding this work we will
be happy to discuss?thow with you.

I Sincerely yours,

  

v. x. 
(fr-inputs.?
.08 I.   
I. I. Intac.rh. -.- . -- 

Acute and Semi?Chronic Toxicity of

Silvex to Laboratory Rats

Biochemical Research Department

The Dow Chemical Company

16335.

 

7?

- ACUTE OF
. .

zl'Biochemical Research'Department
-. The Dow Chemical Company. . .-. .
- Midland, Michigan_ 7 

. Do#;c m?nuv

. 



I'?Silvex" is a trade name that has been?coined for the I

'material known chemically.as propionic' 
-acid. The test suhstance'used in these stddies was a sodium salt 
preparation'in the form_of a brown, rosin-like; hygroscopic solid; 

This material assayed 905 acid equivalent and was shown to con-1:.

tain one molecule of water of hydrationi?



 

1]:f SINGLE-ORAL 

Procedure 

-The_toxicity of Silvex was deternined when adninistered

*Iin singleional doses to yonng adult.white rats of'both sexes from .
Ithe stock colonies of this labofatory; 'AlidUOts of a 10% solu-

xltion of the acid (as the sodium salt) in water were fed hy intuba-'
ition. Eight animals (u o?.each sex)'were used at each of A dosage
levels differing.in quantity.by a geometric ratio of 2. ?All sur- -
vivin? animals were obsenved for about 2 weeks or until they had

fully recOVered from any loss of weight and were gaining normally.

.-

I?esults_
The dosage-reaponse data were treated statistically ac-
cording to the method of "moving-averages? described by_Thompson.
The LD5O 30% rats was found to he 0:50 g./kg; with i9/2o confi?
dance limits of Oi3? t910;65 

. 1

 

 . I .I .

 

?obtained by this lahoratory from the Wistar Institute in 1938.

I ani?els Were maintained-on the stook diet from the time of
iweaning-ontil they were 45 days of age; when they were divided.
?Iaccotding ?toups of?lO rato'of_

feaeh'sei'and.oterted on the control or experimental'diets.

. - 2 

 

DIETARY FEEDING 

W. A I I Procedure

 

. 'All,of.the rats used_in'this work were prOgeny of_stock-



{I'Thefconttolendibaeic retion used in this investigation 

?Wis the usual stock diet employed in this laboretoryui The follow- 

.ing the prep?ration 

I l"?7?nf7?Peroent 9y weight ?iiwt. .
 1' j"    - 



"brie i?xtrad?ed 
:5;Dfied brewers? yeastij?liltf .
G. -looized table Belt-ti. -

.Calcium carbonate' .g5r I 1-OL- -

The experimental diets ?ere ?repareq by thoroughly mixing

the finely ground test material With amounts?

?.Calculated to give diets containing the equivelent of 
0.03, propionic-

acid.

Five reta of the same-sex here caged together in wife
bottom cages_and were allowed free aocess to food and water at
I ,2..r536?1

an

.. n? ., 



I- a 90-day experimental feeding period. Records Were kept of body 2

?then moribund and examined in an.effort to ascertain the cause

'tion.

1.0 Percent"'

. for the control femalcu.

- 

3 all times. All of_the rats were'weighed twice a week for the

first 30 days, and weekly thereafter throughout the remainder of?

weight, general appearance, and average daily food consumption

per animal. Whenever possible, failing'animals were sacrificed

of impending death.

At the end of the 90-day experimental period, representa-

tive groups of female rats were selected for hematological examina-

All of the surviving rats were then fasted overnight, 
neighed, killed_hy decapitation, and examined.. The lungs, heart,
liver, kidneys, and spleen-from all the rats and the testes from _ls
the_male rats were sections off'l?L
these organs,_as well as from the pancreas and adrenals, were: I



prepared 

Results

The diet containing the equivalent of 1% Silvex was not.

- acceptable to the grOUPs.of male and female rats that received

it. Food consumption for each grams per rat
per day conpared to'15.0 g. for the control male rats and 1233 g.

. I - . beeame?rohghland-unkempt"

in appearance, The?average'bodv weight records (see charts) showed

a severe depression. .Within 32 days six of the ten male rats and

. Itng or the 'ten' female rats?had died.? At'jthisl' time, thel?surviving 



.-


Moo__?7?

[15363-

 

were sacrifited and examined. Histopathologic findings

were as ?ollows:
Male and Female Rats,

_f_Heart - Cloudy swelling of muscle fibers. 
'Liver - Generalized cloudy swelling and granular degenera-
tion of parenchyual cells together with foci 
. necrotic cells especially portally.. 

Kidney.? Cloudy swelling of tubular epithelium and vacuole-

tion or epithelium cells of the convoluted (proximal
and distal) tubules._

?stats; "?09

Male Rats.

Testes - Degeneration of the tubules with necrosis of."
germinal epithelium. .



Histopatholngical examination of tissues from the control

animals revealed no abnormalities.

0.3 Percent

'_The groups of rats that received the diet containing

Silvex for 90 days exhibited a marked retardation of growth._

Food consumption records indicated that the animals were accepting
the ration but eating 5115n31- less (15.1 gl/rat/day for the males;
10.9 g. for the femaleel than the animals redeiving the control" I
diet. I - I

Four of the ten sale rats and three-cf the ten female.

rats failed to survive the 93-day experimental?period. These 

deaths were believed to be :ue?to spontaneous infections of-the

lungs or middle ear. The incidence of this type of illness was-

similar in the control and experimental groups.'*

'jvisses



 

 

 .31

II
I.

l" 
IL: 
 

.
.O?Dhcu? -- t- t-

.- 

J. 

pathologic examination revealed:

However) the male rats ex- 

I
gun-

-5-

rerminal hematological values were normal (Table 1).

Organ weight studies (Table 2) revealed a large increase in the




erage weight of the livers and kidneys for both sexes. .Histo-'

Hale'and Female Rats.

172.9461. Moo

Heart - Slight to moderate fatty degeneration of the Enscle
.. fiber. 

Liver - Marked generalized cloudy swelling of the 
. cells. Foci of necrosis in the portal areas involv-
5"ing 3 to 12 cells. 

Kidney Degeneration of numerous uriniferous tubules.

- Vacuolation and cloudy-swelling of the epithelium
cells of many remaining convoluted and collecting
tubules with accumulation of debris in their lumen
but without cast formation. Degeneration of.several 
glomeruli.. - 



Testes Numerous degenerated tubules and degenerated germinal? 
- ~epithelium cells in other tubules but_there was still'
?.evidence of I

a Percent 

. . ,The groups of male and female rats maintained for 90 days

on the.diet.containing 0.1% Silvex seemed normal as far as gross 

. ?Iappearance, behavior, hematOIOgical values (Table and food

Exhibited a moderate depression of growth_(see chart). -A suggestive,
bbut_not statistically significantl depression of growth was ob-
'served for theifemale rats; "Liver?and-kidnev weights were in- .

'Hcreased'significantly.f Histopathologic examination revealed*=153?57



 

Male and Female Rats.

Heart - very slight cloudy swelling and fatty degeneration
in some of the animals.

Liver Marked generalized cloudy swelling of the 
cells. Foci of necrosis in the portal areas involv?
-ing 3 to 12 cells. -

'Kidney Degeneration of numerous uriniferous tubules.
vacuolation and cloudy swelling of the epithelium
cells of many remaining convoluted and collecting
tubules with accumulation of debris in their lumen
but without cast formation. Degeneration of several
glomeruli. - -

MOU-

0.03 Percent

The group of male rats that received the diet containing
0.03% Silvex exhibited a slight depression of growth, but the fe-
male rats grew essentially as well as the control animals (see

charts). Liver and kidney weights each were increased signifi-

'cantly in.the males, but liver weight only was affected in the

female rats. Microscopic examination of the tissues revealed:
Male and Female Rats.

Liver - Marked generalized cloudy swelling of the 
cells. Foci of necro?is in the portal areas involv?
ing 3 to 12 cells.

Kidney - Degeneration of numerous uriniferous tubules.

ovacuolation and cloudy swelling of the epithelium_
cells of many remaining convoluted and collecting
tubules with accumulation of debris in their lumen

but without cast formation. Degeneration of several
glomeruli. 

0.01 Percent
The groups of male and female rats that received the diet

containing 0.01% Silvex for 90 days showed no evidence of adverse

183711

Jinn-?  [an

.--. . . 
. ?1 

Male and Female Rats. I 

Heart - Very slight cloudy swelling and fatty degeneration
in some of the animals. 

-l-Liver Marked generalized cloudy swelling of the 
cells. Foci of necrosis in the portal areas involv
ins 3 to-l? cells.- . I 

Kidney - Degeneration of numerous uriniferous tubules.
vacuolation and cloudy swelling of the epithelium'
'cells of many remaining convoluted and collecting-
tubules with accumulation of-debris in their lumen

but without cast formation. .Degeneration of sevaral,,
glomeruli. I - - 

"Islam; Mon

0.03 Percent

The'group of male rats that received the diet containing

.0.03% Silvex exhibited.a slight depreSsion of-growth, but the-fe-h
- male rats grew essentially as well as the control animals (see"

. 'charts). Liver and kidney weights each were increased signifi?5

cantly in the males, but liver weight'only was affected in?thel
female rats.- Microscopic examination of the tissues revealed:

Male and Female Rats.-
- Liver_- Marked generalized cloudy swelling of.the --W
cells.? Foci of areas involv?

'a ing 3 to 12 Cells. 

'Kidney'- Degeneration of numerous uriniferous tubules;
?.Vacuolation and cloudy-swelling of the epithelium

- cells of many remnanin: canvoluted and collecting.
.tubules with an emulation uf debris in their lumen
but wicked? can; fur.? Degeneration of several


- 

0 '0 .

-. r_ ..




0.01 Percent

The groups of male and_female rats that received the diet

containing.Q.Ol% Silvex for 90 days showed no evidence of_adverse

l\

. - 

 .

after 32' says .-

7 

4-
eiiects

as judged_by growth (see charts); gross appearancc.and

hehaVior, mortality, or food consumption records. Organ'weights .


(Tahle 2) here normal except for Slight increases in the average? 'gg'
weight of_the liver from the female rats and of the kidneys from a;

3the male rats. Microscopic examination revealed: 
(a)l Hale and Female Rats. 

6 Liver - Very'slight cloudy swelling of some of the 
. - - cells in the periphery of the lobe, together with
I foci of necrosis of cells in a few portal areas. 
Kidney [Male Rats)'? Very slight cloudy swelling and vacuola- 
- tion of epithelium cells of numerous
convoluted tubules together with ac?
."cumulation of debris in the_lumens of -.
their tubules but without case formation.?

Sin-suns 3 

Hale and female rats {10 of each sex per group) were

maintained for 90 days on diets containing the equivalent of 0,0

(control), 053, 0.1} 0.01 percent 

oxy) propionic acid (Silvex)31 The actual test material was in the_

for: of the nonohydrate sodium salt. A similar group of rats that
were offered a diet containing?l% Silvex did not accept the ration
'sa

tisfactorili and this portion of the ekperiment was terminated

Growth was retarded for the male rats at all levels ex? 

Icept.0.Ol%, but for the females retardation occurred only at the

0.3% level. I 
- -Terminal hematogieal values were determined on groups of I
5 female rats each fron'the 0.3 and 0.1% levels'and normal results

a.
were obtainedI I


1- J.

J?cceived dietary concentrations
Weights were increased also for
female rats; the average weights of the liver were increased"

.311 groups, including the 0.01% level, out kidney weights were

CD

a 

Organ weight studies showed significant increases in the

average weig

ts of the livers and kidneys from the male rats that

of or 0.03%. Kidney

the 0.01% male animals. For the



4-9 

levated ohly for the female rats on the 0.3 or 0.1% dictS.

I Histooathologic examinatioh revealedevidenoe of adverse
effects, appeefing'predominantly-in the liver and kidneys, from
rats 0; both sexes ?nd at all levels of dietary concentration,

exceht,that the kidneys from the female rats_fed thejlowest dietary

. concentration were not 

 

 

 1.a?r?f 


.

5 AVERAGE HENATOLOGICAL VALUES FROM FEMALE RATS.
. THAT RECEIVED DIETS CONTAINING SILVEX FOR A
. PERIOD OF 90 DAYS

vetermination

 

. Dif

H4

bq??xi? E1


co). . . 

erential Count

Neutrophiles

Monocytes

.Eosinophiies-

Zeucocytes_~Fi?



-9-

1


inn 1

[1

Average value Per Grouo of 5 Rats

Controi I 0.1%

'8.93

I.  18.8 

13.2  

121.8  .
.-. g'

1.2-
2.6

0.305

3.1;899'
I. 

g' 4.311 



813.8 7 .-
70-.8' . 
I:  Jo?s.?  ..

-8.,57

 

2

  {59.1"

  1'3 . 

. 1.27.?.lpi
. we

ggzl.8.

 



. .
.

- I

l- 

AVERAGE HEMATOLOGICAL VA

r.

FROM FEMALE

RATS

THAT RECEIVED DIETS CONTAINING SILVEX FOR A

PERIOD OF 90

 

31?$x1? 31

. Differential Count .
NeutrophilesA





Monocytes



I .



 

Average

3.93-

13;2--5

h)
.

I-q

'mH




Control

5., 18.8 in:

0.3%

(DP
0 rm 
?n.33cn a) 

  8.29.9 - 
'17.1g71u"



value Per Group of 5 Rats

0.1%

 

7.19.1
.31'3-0




 

I
a
.II$13.35. -



 ?fr?H Jr?riffr

.s  

?f10 

HMO-arc?. 

- - . 2

AVISIAGE FINAL BODY AND FRO-111 3.3m 1153335}
.. RECEIVED DIE-TS FOR A PERIOD 

Percent Ho. Orgy-.1: weighth (55-7100 body? are.) .

in Diet . Rats .-  . .. . 
. . . - . .  (3 Lung: ?ea-mt Livar 3:315:01: '??astc-n .

. I
hu-n?uhr Fm . -
. r?I. . .

 

 

 

 

J,Lf33j359: 6.3i 2.45 ?H7e}62 


. . . . 0

*?263,?jg' 0.'11033 '.al

_10.03 
1

Lb.58 '0.31{1 3.15 ,.'o.75l
0.30 l_ 2.46' gp_72aa







:91

3'1!
?3 .

I =?coh?roll'


0.03f '33 10f_
f-107 



0:66__ 0.36 2.h6 p.59 . 0.36 
0.78 _o.u3 5.72 0.30
'0136' i 4.78. 0:76? T0.2a
if.?196 0.35 -3.19 0.7? '0.29
??1'209 6.71 0.36 2.70%, 0.72 . .o.27

03-4:   
?11 w" '1 an

0.01-to 0.05 7 1;.f 
calbuiatea_by "t9_te5t. 

HF -

    

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

    
   

 

    

 

              

 
   
 

 

 

 

 

 

 

             

 

 

 

 

 

 

 

  
      

 

   

 

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thin
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Iskll

 



LIKLIIVIPHIKLGROUPS oEj"
INED FOR I.u..ua)j.jjkn.I)1

 

 

 

 

 

 

T789L6L M00

1 - 2. Results of 90?Day Dietary Feeding of
I Propylene Glycol Isobutyl Ether Ester
F. -of Silvex (Dowco 171) to Rats. Biochemical

Research Laboratory. The Dow Chemical
Company, Midland, Michigan. ?June 19, 1962

 

 

' 

is 
pound
and
upport
ties. In

excre-
of the
inimals
relative
ence of
11. 
.1 made
resence
'ed that
evels in

gonizes
ethacin
.ethacin
les?h 5,
1e .ene
ifferent

are re-
-mpared

ore and
i ulcera-

iacol., 17

2d,, 146

376.

ity Press.
:5 (1975)
iacol., 19

1armacolIqumMu-Imnumlwan?L-buu-muL-l -

-I-

Hit-443. 

 

 

Toxicology, 9 (1978) 75?86
9 Elsevier/North-Holland Scienti?c Publishers Ltd.

STUDIES ON 2.3 ,7 2 UCED

IMMUNE SUPPRESSION AND DECREASED RESISTANCE T0
INFECTION: ENDOTOXIN HYPERSENSITIVITY, SERUM ZINC
CONCENTRATIONS AND EFFECT OF THYMOSIN TREATMENT

J.G. WOS1 J.G. H.W.B. ENGEL, A. MINDERHOUD and L.M. VAN
NOORLE JANSEN 

National Institute of Public Health, P.O. Box 1, Bilthoven (The Netherlands)

(Received August 23rd, 1977)
(Accepted October 3rd, 1977)

 

SUMMARY

Sublethal doses of the highly toxic chemical 2,3,7,8-tetrachlorodibenzo-p-
dioxin (TCDD) cause thymus atrophy in several species and induce suppres-
sion of cell-mediated immunity as measured by different parameters. The
selective effect on thymus is not likely caused by a cytotoxic effect on
or by an effect on pituitary or adrenals. In the present study,
mice received daily injections with thymosin in order to study whether
reduced production of thymic hormones could be involved in the atrophy.
Also, serum zinc concentrations were measured. As thymosin injections
did not increase thymus weight and mitogenic responsiveness of thymocytes,
and zinc levels were not depressed, the mode of action of TCDD-induced
thymus atrophy remains unknown. It has been reported that TCDD marked-
ly decreased resistance of mice to infection with Salmonella bern. In this
study, data are presented that indicate that the increased susceptibility is
likely due to the endotoxin content of the bacteria: pre-treatment of mice
with single or repeated doses of TCDD markedly enhanced their suscepti-
bility to endotoxin even at dose levels that did not produce thymus atrophy.
Finally, possible effects of TCDD on macrophage functions were studied. As
treatment with TCDD did neither impair non-specific killing and phagocyto-
sis of Listeria monocytogenes, nor macrophage reduction of nitro-blue tetra-
zolium, it seems likely that the immunosuppression is only due to a, to date
unknown, effect on and not due to a combined effect on
both T-cells and macrophages, and that the endotoxin hypersensitivity is not
the result of alteration in phagocytic function of macrophages.

 


(TCDD) is a chemical that can be

75


0



laser.

 

- - - -.  7 
fer:- a 9* I . 

formed in small, but significant, amounts in the production of some chlo-
rinated phenols or chemicals from chlorOphenols, such as the
herbicide 2,4,5,-trichlor0phenoxyacetic acid. It is one of the most toxic
chemicals known it is a teratogen it is associated with chick
edema disease in chickens and with several industrial incidents resulting
in cases of chloracne in man TCDD exposure has been shown to
induce thymus atrophy in rats, mice, guinea pigs and monkeys
[McConnell and Moore, in press]. Thymus atrophy appeared to be one of
the most consistently sensitive parameters of TCDD exposure. Since 
cytes derived from the thymus (T-cells) play a central role in cell-mediated
immunity, several funciton studies of the thymus?dependent immunity have
been carried out in mice, rats and guinea pigs. TCDD exposure did result in
functional impairment of parameters of cellular immunity (delay~
ed type hypersensitivity, allograft rejection, graft-versus?host activity and
responsiveness of to the mitogens phytohemagglutinin (PHA)
and concanavalin A (Con A) [11,13, Moore and Faith, in press] but the
suppression of the cell-mediated immunity mems to be an age-related pheno-
menon, at least in the mouse and the rat where TCDD exposure during the
ontogEnesis of the immune arstem seems to be pre-requisite in order to
obtain immune suppression. TCDD mimics in this respect the effect of
neonatal thymectomy.

Since the TCDD-induced atrophy of the thymus can be explained in
many ways and no clear-cut mode of action has been described, we were
interested to examine whether a reduced production of thymic hormones by
thymus-epithelial cells could be involved. Therefore, in the present study,

atrophy was studied. As shown by Thigpen et al. TCDD exposure
markedly decreased the resistance of mice to infection Salmonella bem,
whereas TCDD has no significant effect on mortality in pseudorabies virus-
. infected mice. In the present study, data are presented that indicate that the
increased susceptibility to Salmonella infection is very likely due to the
endotoxin content of the bacteria. Since on the one hand zinc deficiency can
result in severe atrophy of organs [15,16] and on the other hand
parenteral administration of zinc increases the resistance of mice to endo-
toxin [17] serum zinc concentrations were also measured in TCDD-exposed
mice. Finally as macrophages play an important role in the immune response
and in the detoxification of endotoxin, function studies of macrophages
(non-specific phagocytosis and killing of Listeria monocytogenes and reduc-
tion of nitro-blue tetrazolium by macrophages) were also included.

the effect of daily administration of thymosin on TClD-induced thymus 

MATERIALS AND METHODS
TCDD

(purity 98.6%, Lot No. 851144-11"Manama?Mb 

15382,

a)

  

?me chlo-
:h as the
351: toxic
ith chick
resulting
hown to
monkeys
ne one of

nediated
iity have
result in
(delay-
vity and
(PHA)
but the
:l pheno-
iring the
order to
-ffect of

ained in
we were
ion/WY
st 
thymus
xposure
'la bern,
as virus-
:hat the

to the
ncy can
er hand
endo-
xposed
esponse
Iphages

reduc-

4441,

?annulus?ad I. In. v-u p-uvu Jim.?

I .Il. "In

 

nuance-4 an aim ?I-lbl'n?

.-.. .. L.-. .I.

I hula



 

kindly provided by Dow Chemical Company, Midland, Mich., U.S.A.) was
dissolved in reagent grade acetone at a concentration of 50 pg/ml. For the
animal experiments, this stock solution was diluted with at least 9 parts of
arachis oil to provide the different concentrations. Mice were intubated
orally with 0, 1.body weight. 

Animals

Speci?c pathogen-free outbred Swiss mice were reared at the Institute.
They received food and water ad libitum. Because of the high toxicity of
TCDD, all animal experiments were carried out under non?sterile conditions
in plastic Trexler isolators, and all contaminated material was incinerated at
temperatures over 800? (N.V. Roteb, Rotterdam, The Netherlands).

Zinc analysis

For the determination of zinc in serum, an atomic absorption Spectro-
meter with acetylene-air slot burner and zinc hollow-cathode lamp was used.
Fifty pl of serum was diluted with 500 pl of distilled water. After mixing
thoroughly 100 pl of the sample was aspirated into the ?ame and the result-
ing atomic absorption signal was recorded. Peak heights were compared with
results obtained from the aspiration of aqueous standard solutions under the
same conditions and zinc concentrations were calculated.

Experiment with thymosin

The effect of thymosin injections on TODD-induced thymus atrophy was
studied in mouse pups that were exposed to TCDD by postnatal maternal
treatment. At birth the number of neonates was standardized to 6. Groups
of 5 nursing mice received 0 or 10 pg body weight (in a volume of
0.1 ml/3O postnatally on days 1, 4, 8, 11, 15 and 18. Calf thymosin
(fraction 5, Lot No. BPM 390, kindly provided by Dr. A.L. Goldstein,
Galveston, Tex., U.S.A.) was dissolved in sterile saline at a stock concentra-
tion of 15 mg/ml, passed via Millipore bacteria filter and aliquots stored at
?20? C. Just prior to injection, a dilution (1 mg/ml) was made in sterile
phosphate-buffered saline (PBS). In the first week, groups of 2 pups per
litter received daily subcutaneous injections of a high (50 pg) or low (15 pg)
dose of thymosin. In addition, 3 pups per control litter received PBS. In the
second and third week, the doses were doubled and tripled respectively. The
endotoxin content of the thymosin (1 mg/ml) solution was estimated with
the Limulus assay [18] and contained between 12.5 and 62.5 ng endotoxin/
ml. After the 22-day experimental period, the animals were killed.Zinc concen-
tration was measured in serum. 

The thymus of half of' the number of mice was removed under sterile
conditions. Cell suspensions were made by using a glass homogenizer, the
viability was determined with _a dye exclusion test and the final suspension

77

9819890M00

 

18383

was adjusted to a concentration of 3 10" viable nucleated cells/ml in
minimum essential medium (MEM, Gibco F14) buffered with Tris (0.025 M)
and supplemented with 20% inactivated fetal calf serum, streptomycin (100
ug/ml) and penicillin (100 lU/ml). A total volume of 1 ml was cultured in
tubes (Nunc, Denmark) and 12.5 ul reconstituted phytohemag~
glutinin (PI-IA, Wellcome Labs., Beckenham, Kent, U.K.), 6 ug concanavalin
A (Con A, Pharmacia, Uppsala, Sweden) or 50 ul reconstituted pokeweed
(PWM, Gibco) were added. After 24 cultivation,10 ?Ci of [6-3H1thymi-
dine (specific activity 600 mCi/mmole) was added. Twenty-four hours later
the cells were harvested on glass fiber filters using a multiple cell culture
harvester (Skatron, Lierbyen, Norway). After drying, the filters were placed
into scintillation vials and 2 ml of scintillation fluid were added. Counting
was done in a Packard liquid scintillation counter. Results (median value of
triplicate cultures) were expressed as the difference between stimulated and
non-stimulated cultures (in counts/min (cpm) per 3 cells).

Experiments with endotoxin

In 3 experiments, the effect of TCDD on the susceptibility of mice to
endotoxin was studied. Since there is no major difference in the biological
activity of endotoxins of Salmonella and E. coli species, B. coti endotoxin
was used in the present study. Endotoxin (iipopolysaccharide, E. coli, 0
127: 8, Difco Labs, Detroit, Mich.) was dissolved in sterile saline. In two
studies, 3?4 week-old male Swiss mice were intubated orally, once a week
for 4 weeks, with 0, 1.5, 5, 15 or 50 ug (0.1 nil/10 body weight).
Two days after administration of the final dose of TCDD. the mice were
injected intravenously with 10, 20, 100,250 or 500 pg endotoxin in 0.1 ml.
The mortality due to endotoxin shock was scored after 48 h. Surviving
animals were killed and examined macroscopicaiiy.

In the third experiment, 3?4 week-old female mice received a single oral
dose of 0 or 100 ,ug body weight (0.2 ml/10 Animals were
challenged with endotoxin 5 days after intubation. In the second and third
experiment, additional animals were intubated and killed to determine the
effect of TCDD on body weight and on the weight of thymus and spleen.
Also, serum zinc concentrations were measured in the third experiment.

Clearance of Listeria monocytogenes

The resistance of Listeria monocytogenes is a combination of non-specific
phagocytosis and cell-mediated immunity: non-specific phagocytosis and
killing can be measured shortly (within 2 days) after the intravenous inocula-
tion of Listeria monocytogenes Male Swiss mice (3?4 weeks old) were
intubated orally, once a week for 4 weeks, with 0 or 50 pg body
weight. Four days after the last intubation the animals were inoculated intra-
venously with 2.4 10" Listeria monocytogenes organisms (Strain 242/73
type 4b). One or two days after the injection. the spleens were weighed,

78

1638?



 

I
.-
--
I
.

 

  

Ills/ml in
0.025 M)
min (100
iltured in
tohemag?
canavalin
'okeweed
l-l]thymi-
ours later
.1 culture
re placed
Counting
1 value of
lated and

mice to
)iological
ndotoxin
coli, 0
e. lip?two
a\ 
weight).
Lice were
:1 0.1 ml.
Surviving

ngle oral
lalS were
Ind Ulkd
mine the
spleen.
eriment.

I-specifit'
osis and
inor-ula~
lid) were
kg body
ed intra-
.242573
.veighed,

 

- nan-am

?05 nuns-doll! I to 



'u-I-Mnu-u n- - -.

'dld I Mumn?unumn

i?l? ~51. lay-a.

g.

. I 

n- A

 

homogenized and serial dilutions were plated to determine the viable counts
of Listeria The number of bacteria was expressed as mlog/spleen.

Reduction of nitro-blue tetrazolium

activated macrophages show increased glucose oxidation via
the hexose-monophosphate pathway; an increase that can be quantitated by
the reduction of soluble nitro-blue tetrazolium (NET) to insoluble formazan.
Krueger et al. [21] demonstrated a correlation between delayed type hyper-
sensitivity skin tests and NET reduction by macro-
phages. Similarly Engel and Sekhuis (unpublished data) showed an increase
in NET reduction in macrophages of mice that were injected with Coryne-
bacterium parvum, without the addition of ly'mphokines?to the culture
chamber. In the present experiment, the effect of TCDD on NET reduction
of macrophages was determined in order to study an effect on the baseline
activity of non-activated macrophages. For that purpose, 3-4?week-old male
Swiss mice were intubated orally, once a week for 4 or 5 weeks with or 50
pg body weight. Five days after the last intubation, peritoneal
macrophages were harvested in 2 washings with 2.5 ml Eagle's Basal Medium
(Gibco, 13) supplemented with 20% inactivated calf serum, 40 mM

L-glutamine and 5 heparin/ml. Of this suspension 0.5 ml was added to an -

8-chambered tissue culture slide and incubated this time chambers were washed 3 times to remove non-adherent

cells. Of each mouse, one chamber was used for the identi?cation of macro-
phages (Giemsa stain); the other was used for the NET assay according the
technique described by Krueger et al. [21] . One hundred cells were indi-
vidually scored at 1000 on the quantity of formazan precipitate as a
measure of NET reduction (no precipitate 0; slight precipitate 1; mode-
rate precipitate 2; strong precipitate 3). Results were expressed as the
sum of the scores per 100 cells.

Statistical analysis

Student?s test was used to make one-sided treatment?control compari-
sons.

RESULTS

The effect of daily thymo?sin injections on body and thymus weights of
22day-old mice is given in Table I. Body and thymus weights of the 3 TCDD
groups were significantly lower when compared with the control group.
Treatment of TCDD-exposed pups with thymosin significantly reduced body
weight, but did not alter thymus weight. Serum zinc concentrations were the
same in control and TCDD?eXposed pups. The responsiveness of thymus cells
to the mitogens PHA, Con A and PWM is presented in Table II. In general,
the response of the TCDD groups to these mitogens was approximately half

79





 

TABLE I

BODY AND AND SERUM ZINC CONCENTRATIONS OF 22-

DAY-OLD TCDD-TREATED MICE THAT RECEIVED DAILY INJECTIONS WITH
-

 

 

Dose of TCDD Thymosin Body weight Thymus weight Zinc
(uclks) treatment (2) (me) (mail)

0 0 1312.4 0.11b 60 10c 1.2 1 0.5 
10 low doses? 11.2 2 1.0Ml so 2 10?
10 high doses? 11.2 2 53 2 7c .5

 

Mean values S.D. (body weight 8?10 animals, thymus weight and zinc analyses of 
4?5 animals). Mice were expOeed to 0 or 10 pg by maternal treatment post- Co
natally on days 1,4, 8. 11, 15 and 18.

Signi?cantly (P 0.05) different from control group (no TCDD and no thymosin).

(P 0.001) different from control gmup (no TCDD and no thymosin).

Signi?cantly (P 0.025) different from thymosin control group (10 ug TCDD and no C70
thymosin).

For schedule see text.

[9

8

of the response seen in the control group. Since the results are expressed on
a cell-for?cell basis and thymus weight was reduced by TCDD treatment, the
. total reSponse per thymus is likely even more decreased. Thymosin treat-
ment of TCDD-exposed mice did not result in a signi?cant increase in the
mitogenic responsiveness of the thymus cells.

The results of the ?rst two experiments with endotoxin are given in Table
Pretreatment of mice with TCDD markedly enhanced their susceptibility

TABLE II

MITOGENIC RESPONSE OF THYMUS CELLS FROM 22-DAY-0LD TCDD-TREATED
MICE THAT RECEIVED DAILY INJECTION WITH 

 

 

Dose of Thymosin A FHA A Con A A PWM
TCDD treatment (cpm) (cpm) (cpm)
(Milk!)

0 0 19,001 2 10,774 20,895 6,517 6,494 2,647
10 0 3,097 2,731b 10,779 6,345b 2,103 1 1,379b
10 low closes? 9,985 8,248 14,616 1 4,989 3,714 1,485
10 high doses? 11,373 1: 4,677 11,244 2,595b 2,542 1,135?

 

Mean values 2 S.D., 4?5 animals per group. Mice were exposed to 0 or 10 ug 
by maternal treatment postnatally on days 1, 4, 8, 11, 15 and 18. Results are expressed
an the difference between stimulated and non-stimulated cultures (in cpm/3 10? cells).

5 Significantly (P 0.05) different from control group (no TCDD and no thymosin).

For schedule see text.

80

16383

 

AI-thnl?n a" 1

 

 

 

  

01" 22-
WITH

Zinc
(mall)

I. It

Hr?
NH

inalyses of
men: post-

103i?).
nosin).
DD and no

ressed on
her?" the
sin?a 
se in the

in Table
aptibility



 

2,547
1,2379h
1,485
1.135?

 


expressed
10? cells).
~sin).

-b-uhuo-u-nu-l- new.

"drawn-Illnedo?Mu

f4.

 

.ll-I??q-h?MI .-

 

TABLE 

EFFECT OF 4 WEEKLY DOSES OF TCDD ON THE 0F MICE T0
ENDOTOXIN

 

Dose of TCDD Incidence of mortality?

(us/kg)? 
10 pg endotoxin 250 ug endotoxin

 

 

0/4 0/5

1.5 0/4 1/5

5. 0/4 6/endotoxin 100 ug endotoxin 500 ug endotoxin

 

0 0/6 0/6 2/two experiments, male mice (3-4 weeks of age) were intubated orally, once a week
for 4 weeks, with TCDD in acetone-arachis oil carrier.

Intravenous injection with endotoxin (E. coli. LPS) two days after last intubation. Mor-
tality scored after 48 h.

ND not done.

to endotoxin. After the injection of 10 pg endotox'm, 2 out of 4 animals
died in the 50 pg TCDD group and 1 in the 15 ng TCDD group. Injection of
250 pg endotoxin was lethal for all mice in the 5 and 15-pg TCDD groups
and caused one death in the 1.5 ug TCDD group, whereas all controls surviv-

TABLE IV

EFFECT OF 4 WEEKLY DOSES OF TCDD ON BODY AND ORGAN WEIGHTS OF


 

 

Dose of TCDD Body weight Thymus weight Spleen weight
018/?) (mg) (ma)

0 30.7 2.2 66 20 105 13

1.5 2326.0 2.3c 34 10c 89 2 ab

 

Mean values S.D., 6 animals per group. Male mice. 3-4 weeks of age, were incubated
orally, once a week for 4 weeks, with TCDD in acetone-arachis oil carrier.

0.025. 

0.005.

81

 

BEIQBQOMOU

15387

 

TABLE 

EFFECT OF A SINGLE ORAL DOSE OF TCDD ON BODY AND ORGAN WEIGHTS
AND ON SERUM ZINC CONCENTRATION OF AND ON THE SUSCEPTIBI-
TO ENDOTOXIN -

 

 

Dose of TCDD Body weight3 Thymus weight? Spleen weight" Zinca
(us/kg) (2) (mg) (mg) (mail)

0 19130.5

 

Incidence of mortalityc 

 

20 pg endotoxin 100 pg endotoxin 500 pg endotoxin

 

Mean value S.D., 7 animals per group. Female rniceI 3?4 weeks of age, Were intubated
once orallywith .TCDD in acetone-arachis oil carrier and killed after 5 days.

I:
0.05.

Intravenous injection with endotoxin (E. coli, LPS) 5 days after intubation. Mortality
scored after 48 h.

ND not done.

ed. In the second experiment, animals were given 0, 1.5 or 50 pg 
body weight and were challenged with 10, 100 or 500 pg endotoxin. The
10 pg dose of endotoxin caused 2 deaths out of 5 animals in the 50 pg
TCDD group, and a similar mortality (2 out of 6) was scored in the controls
injected with 500 pg endotoxin. Thus, the susceptibility to endotoxin was
approximately 50-fold enhanced in the 50 pg TCDD group when compared
with the controls. Even at the 1.5 pg TCDD level there was a increas-
ed mortality after injection of endotoxin. This latter dose of TCDD being
more than one order of magnitude lower than the dose that caused thymus
atrophy at gross inspection. As shown in Table IV, body weights and weights
of thymus and spleen of mice of the 50 pg TCDD group were significantly
decreased, whereas these parameters in the group intubated with 1.5 pg
body weight did not differ from the controls. A single oral dose
of 100 pg body weight signi?cantly reduced thymus and spleen
weight of female mice, killed after 5 days; but body weights and serum zinc
levels were not altered (Table V). Also, this treatment enhanced the suscepti-
bility to endotoxin: a 100% mortality occurred in the TCDD-exposed
animals after injection with 20 pg LPS, whereas a similar mortality was seen
in controls receiving 500 pg LPS.

The results of the study of the effect of TCDD on non-specific phagocyto-
sis and killing ofL. monocytogenes are given in Table VI. The number ofviable
Listeria organisms, determined One or two days after inoculation. of this

82

16388

 





 

 

GHTS


m?

2/1)

 

I.
.09
IBM

 

 

atoxin

 

 

ubated

vrtality



)D/?kg

t. The
50 ug
ntrols
was
pared
creas-
being
ymus
eights
antly
-5 us
close
pleen
zinc
:epti-

.seen

cyto-
Iiable
this

nd--n.

uAln-M 

ML.

 

nun-no wanna-dd- hb-uu Mb - . 

 

TABLE VI

EFFECT OF 4 WEEKLY DOSES OF TCDD IN MICE ON BODY AND SPLEEN
WEIGHTS AND ON DAY 1 AND DAY 2 SPLEEN COUNTS OF LISTERIA


 

 

of TCDD Body weight. Spleen weight Listeria countb
(Jig/kg) (2) (mg) ("log/spleen)
Day 1 p.i.

0 23.4 1.4 140 10 4.89 0.40
50 . 21.7 0.6c 96 2 13" 4.53 0.24
Day 2 p.i. .

0 21.7 1.5 178 23 5.13 0.31
50 21.2 a 2.4 124 29d 5.03 0.27

 

Mean values 8.0.. 5 animals per group. Male mice.3?4 weeks of age, were intubated
Orally, once a week for 4 weeks with TCDD in acetone-arachis oil carrier.

Animals were inoculated intravenously with 2.4 10? Listeria monocytogenes organisms
4 days after the last intubation; the number of viable Listeria organisms per spleen was
determined after 1 and 2 days-

0.05.
0.01.

organism. was the same in spleens from treated and control mice. Spleen
weights were significantly lower in TCDD?exposed animals. In addition,
body weights of mice killed one day after inoculation were signi?cantly
reduced in the TCDD group. Finally, as shown in Table VII, in two experi-
ments TCDD did not alter the number of peritoneal macrophages, nor did
it cause a significant effect on the macrophage reduction of NBT.

TABLE VII

EFFECT OF 4 OR. 5 WEEKLY DOSES OF TCDD IN MICE ON NUMBER OF PERI-
TONEAL MACROPHAGES AND ON MACROPHAGE REDUCTION OF 

 

 

Dose of TCDD No. of peritoneal Reduction of
(pg/kg) macrophages (x 10?) 

Four doses

0 1.94 -. 0.68 32.2 22.2
:30 2.23 0.35 37.0 8.9
Five doses

0 23.8-1.l
50 2.08 -. 0.3l 57.0 0

 

Mean values - S.D.. 4?5 animals per group. Mah- mite, Weeks of age, Were- intubat-
o-d orally, a Week for 4 or 5 weeks with TCDD in metune oil carrier.
For details see text.

83

1719890MOU

16383

 

DISCUSSION

From the present results it can be concluded that thymOSin treatment of
mice at the employed doses does neither protect the animals from thymus
atrophy nor from a reduced mitogenic responsiveness of thymocytes. The
presence of a minor quantity of endotoxin in the thymosin solution could
not conceal a possible increase, since treatment of mice with E. coli LPS
has been shown to increase the PHA and Con A responsiveness of thymo-
cytes [22] . Thus, it seems unlikely that the depletion of in the
thymus, as induced by TCDD, is caused by a reduced production of thymic
hormones, since calf thymosin has been shown in several studies to potenti-
ate various parameters of thymus-dependent immunity Neither seems
zinc de?ciency to be a cause for the thymus atrophy. A direct cytotoxic
action of TCDD on is also unlikely, since the mitogen respon-
siveness of mouse or rat was not reduced when cultured in the

presence of TCDD at concentrations up to 0.02 Jug/ml medium Even .

the presence of 0.32 ,ug medium has been reported to be not toxic
for human [24] whereas concentrations that are two orders of
magnitude lower do induce aryl hydrocarbon hydroxylase activity in human
[25] . In another study [26] a cytotoxic action of TCDD at a
concentration of 0.48 .ug/ml was reported for several mammalian cell types.
These results make it unlikely that TCDD does need metabolic activation,
but they do indicate that the depletion of the thymus and the
thymus-dependent tissue are not caused by a direct effect on
but rather by an indirect way. There are several conditions
that can cause thymus atr0phy [27] . Regarding the selective effect of TCDD
on thymus, it is not likely caused by an effect on pituitary, on adrenals or
by reduced food intake, since thymus atrophy did occur in TCDD-exposed
rats that were hypophysectomized or adrenalectomized and in rats that were
placed on a restricted diet [Van Logten, Gupta and Moore, in preparation]
though it is still possible that TCDD may impair the absorption and trans-
port of certain nutrients. In addition, serum a-fetOprotein levels are not
elevated in TCDD-exposed rats [Vos and Boekestein, unpublished data],
thus the observed immunosuppression is not due to a a-fetoprotein induc-
tion. In conclusion, the mode of action of the TODD-induced thymus
atrophy remains unknown. 

The results of the present experiments with endotoxin show a marked in-
crease in susceptibility to endotoxin of mice treated with a single or with
repeated doses of TCDD. This sensitizing effect of TCDD for endotoxin
offers a logical explanation for the decreased resistance of TODD-exposed
mice to infection with (endotoxin-containing) Salmonella bacteria as observ-
ed by Thigpen et al. [14] . Another explanation for the increased suscepti-
bility to this type of infection, which is primarily dependent upon cell-
mediated immunity [28] is by general suppression of the thymus-dependent
immunity. This explanation is, however, less likely since increased mortality
by Salmonella occurred at dose levels of TCDD that did not produce thymus

84

 

..- .- -. . -.

isato

 

 

ment of
thymus
:es. The
could
-oli LPS
thyme-
as in the
thymic
potenti-
ar seems
ztotoxic
respon-
?d in the
Even
ot toxic
rders of
1 human
DD at a
ll types.
ivation,
and" "te
'fec .n
iditions
TCDD
anals or
exposed
at were
ration]
i trans-
are not
data] 
induc-
:hymus

'ked in-
31' with
lotoxin
xposed
observ-
iscepti-
in cell-
endent
)rtality
hymus






4 I

mil-l an noth I I4 r- 

min I .- Ipuld?v 

-I.- -.

gnu-us la

 

atrophy. The endotoxin hypersensitivity, which has to date only been studi-
ed in mice, is a phenomenon of particular interest when considered in
connection with: (1) terminal hemorrhages that were seen in TCDD-exposed

rats and guinea pigs [8,29] and mice [12,30], (2) thrombocytopeniav

observed in rats and guinea pigs [10,31], and (3) presence of thrombi in
blood vessels of heart and lung in the rat after a lethal dose of TCDD 
These lesions are also seen in endotoxin shock and since TCDD, at least in
the mouse, also induces endotoxin- hypersensitivity, one has to consider
endotoxin shock as a possible cause of death. Regarding the mechanism of
endotoxin hypersensitivity, several possibilities exist (see review of Cook
et al. With regard to the endotoxin hypersensitivity produced by
TCDD it would be to investigate the role of the enzyme gluta-
thione-S-transferase B, a cytosolic glutathione-conjugating enzyme, since
sul?'iydryi containing enzymes are necessary for endotoxin inactivation.
TCDD has been reported to increase the activity of both hepatic and renal
glutathione?S-transferase [33] .

Finally, as treatment of mice with TCDD did neither impair the non-
speci?c killing and phagocytosis of L. monocytogenes nor the macrophage
reduction of NBT, it seems likely that the TCDD-induced suppression of
the cell-mediated immunity is only due to a, to date miknown, effect on
T-Iymphocytes and not due to a combined effect on both T-cells and macro-
phages. Similarly, the results of the experiment with L. monocytogenes
indicate that the endotoxin hypersensitivity is not the result of alteration
in phagocytic function of macrophages but, as discussed in the previous
paragraph, perhaps due to impaired endotoxin detoxifying preperties of
macmphages.

REFERENCES

1 B.A. Schwetz, J.M. Norris, G.L. Sparschu, V.K. Rowe, P.J. Gehring, J.L. Emerson and

QC. Gerblg, Environ. Perspect., 5 (1973) 87.

KB. Courtney and J.A. Moore, Toxicol. Appl. Pharmacol., 20 (1971) 396.

D. Neubert, P. Zens, A. Rothenwallner and H.J. Merker, Environ. Perspect., 5

(1973) 87.

D. Firestone, Environ. Pempect., 5 1973) 59.

Crow, Trans. St. John's Dermatol. 56 (1970) 79.

J. Kimmig and K.H. Schulz, Dermatologica, 115 (1957) 540.

N.P. Buu-Hoi, RH. Charm, G. Sesqu?, M.C. Azum-Gelade and G. Saint-Rut, Natur-

wisa., 4 (1972) 174. 

8 EN. Gupta, J.G. Vos, J.A. Moore. J.G. ?link! and V. Bullock, Environ. Per-
spect., 5 (1973) 125.

9 M.W. Harris, J.A. Moore, J.G. Vos and B.N. Gupta, Environ. Perspect., 5 (1973)
101.

NM



10 Kociba, RA. Keeler, C.N. Park and P.J. Gehiing, Toxicol. Appl. Pharmacol., 35

(1976) 553.
11 J.G. Vos, J.A. Moore and J.G. Zinkl, Environ. Perspect., 5 (1973) 149.
12 J.G. Vos, J.A. Moore and J.G. Zinkl, Toxicol. Appl. Pharmacol., 29 (1975) 229.
13 J.G. Vos and J.A. Moore, Int. Arch. Allergy Appl. lmxnunoL, 47 (1974) 777.

85

0471 9890M00



 

J.E. Thigpen, R.E. Faith, E.E. McConnell and J.A. Moore, Infect. Immun., 12 (1975)
1319. 

W. Groth, Zbl. Vet. Med. A, 23 (1976) 31.

BL. O'Dell, P.M. Newberne and J.E. Savage, J. Nutr., 65 (1958) 505.

Snyder and FLI. Walker, Infect. Immun., 13 (1976) 998. 

J.G. Kreeftenberg, H.G. Loggen, J.D. van Ramshorst and EC. Beuvery, Develop.
Biol. Stand., 34 (1977) 15.

S.P. Tripathy and GB. Mackaness, J. Exp. Med., 130 (1969) 1.

E.J. Ruitenberg and L.M. van Noorle Jansen, Zbl. Bakt. Hyg., I. Abt. Orig. A, 231
(1975) 197.

G.G. Krueger, B.E. Ogden and Weston, Clin. Exp. Immunol., 23 (1976) 517.
L. Adorini,- L. Ruco, S. Uccini, G. Soravito de Franceschi, C.D. Baroni and G. Doria,
Immunology, 31 (1975) 225.

A. White, Thymus hormones: a new ?eld in endocrinology and its signi?cance fOr
host immunological competence. In: D.W. van Bekkum Activity of Thymic
Harmanes, Kooyker Scientific Publications Rotterdam, 1975, p. 17.

P.W. Beatty, E.J. Lembach. M.A. Holscher and R.A. Neal, Toxicol. Appl. Pharmacol.,
31 (1975) 309.

B.E. Kouri, H. Ratrie, S.A. Atlas, A. Niwa and D17. Nebert, Life Sci., 15 (1374)
1585.

A. Niwa, K. Kumaki and D117. Nebert, Mol. Pharmacol., '11 (1975) 399.

J.G. V05, Crit. Rev. Toxicol., 5 (1977) 67.

KM. Collins and GB. Mackaness, J. Immunol., 101 (1968) 330.

K.S. Khera and J.A. Ruddick, Adv. Chem. Set. 120 (1973) 540.

G. Jones and J.B. Greig, Experientia, 31 (1975) 1315.

J.G. Zinkl, J.G. Voa, J.A. Moore and B.N. Gupta, Environ. Perspect., 5 (1973)
111.

J.A. Cook, N.R. DiLuzio and E.O. Hoffman, Crit. Rev. Toxicol., 3 (1975) 201.

R. Kirsch, G. Fleischner, K. Kamisaka and LM. Arias, J. Clin. InVest., 55 (1975)
1009.

 



1,63"?

 

. 2438

Lennart Hardell*:

Malignant tumors and exoosure to 35
phenoxy acids_? A clinical observation [Haligna mesenkvmala 5:
tumorer och exposition for Eenoxisyror - en klinisk obsor? C3
vation]. kn
Lakartidningon, 14:, (33), pp. 27s*?2754, 1977*Dept. of Oncology. Regional Hospital, Umea. we

01

is

CH

A


DEPARTMENT

 

INDEX

M. J. Traynor - 2030

   
   

 

B. B. Holder - 2030
H. C. Scharnweber - 2030
R. J. Kociba - 1803
- B. A. Schwetz - 1803
P. J. Gehring - 1803
v. K. Rowe - 1803 
J. G. Gleason - 2030
V. 8. Robinson IN
C. Spencer - A
Li H. L. Lenq
D. McCollister
Jom1H.Da?duun 11/23/77

 

Translated from Swedish by the Ralph Hc?lroy 00., Custom DiVlSiOn,
2102 Rio Grande, Austin, Texas 73705 USA

16393

Code: 101-8715

MALIGHANT TUHQRS AND EXPOSURE TO
PHENOXY ACIDS A CLINICAL OBSERVATION

Sewn cases of main-Hum lu-
mnurx u! the dept of Lincolngy in
Una-:5 during :21: yur: PERL-76 and ?uh capo. 
sun- lo acids user pcnou? of 
ycurs an: Th: txrmurc was diam:
and mam?; in the of the
cast-s. Thu period 1.5 in agreement with
that assumed fur The
dixniiruliml tin: status in (his maiminl.
of 57 path-uh in all. deviates from 
avtragc. with a dominance of males. Nu deli-
nit: conclusiuns tenccmin: .1 possible causal
connexion can be drawn from caws:
this would can for men: extensive 


896250Nu

svgtegomon

The possibility that phenoxy acids (for example, hormoslyr)
and the contaminants that occur in them may have harmful effects
in humans and animals has been the object of an intensive debate.
No cases of carcenogenesis in man have been reported so far.

In this article we shall describe seven patients suffering
from malignant tumors observed at the oncological
clinic in Umea during the years 1970?76. In these patients the
exposure to phenoxy acids had occurred 10?20 years before the
diagnosis of the tumors. These cases do not, naturally, prove
any possible connection between such an exposure and the malignant
tumors; to prove or disprove such a connection extensive epi?
demiological studies are required. However, we believe that
adequate evidence is available to report the observations in
a casual form.

During the years 1970-76, the oncological clinic in Ume;
received (and checked) 87 patients who had malignant 
tumors. 32 of these were women (372) and 55 were men 43
of the men have been employed; occupation is missing, however, in

I

one case. Nine were forestry workers four worxed in

farming and forestry and six in saw mills or the pulp indus-

vtry 

1.6334



Case 1. A 62 year old man, forest worker until retirement in
1971; presenting diarrhea and pains on the left side of
the stomach. Smoker. Treated for tuberculosis in 1959. Other?
wise, previously basically healthy. In this work he had sprayed
the phenoxy acid roughly one week in 1963 and 1964,itwo
weeks in 1965, one month in 1966, one month in 1967, and two weeks
in 1968. .Since August 1976 increased pain in the right fossa
iliaca and also swelling in the right leg. Laparotomy revealed

a tumor in the region of the small of the back, attached to the
wall of the hip bone. DIAGNOSIS: Rather highly differentiated



leiomyosarcoma.

Case 2. -A 57 year old forest worker, nonsmoker. High blood
Ipressure since 1960. Treated for postinfectious arthritis in
1974. In the 1950's and 196 for three or four summers each,
and for each summer for three weeks he had worked using a mixture
of and for spotting and to a lesser extent for back
spraying. Sinde August 1976 ventral tumor growth, on the proximal
portion of the right thigh. DIAGNOSIS: tumor, pro-

bably relatively highly differentiated fibroid liposarcoma.

Case 3. A 49 year old man, workshop owner. For many years gastric
but otherwise healthy. On his workshop premises, from
1961 to 1972, he had stored thousands of liters of phenoxy acids
for a forestry company during the nonspraying season, about 11
months per year. The acid was stored partially in open containers.
The patient had come in contact with the substance, which emitted
a constant odor in the shop. In August 1976 melena and_stomach
pains. In November again acute stomach which led to
laparotomy. This revealed a tumor in the region of the Shall of
the back and involvement of the rectum, urinary bladder and over-
growth to the small intestines. DIA NOSIS: tumor,

more specifically, neurosarcofibroma.

16395

. 

Page 3

Casg_i. A 60 year old forest worker, moderate smoker. In 1938
appendectomy, but otherwise healthy. During the years 1961-66
for 20 workdays per year he had sprayed with phenoky acids.

In August 1971 saw a doctor for defecation trouble. Palpation

. C3

of the rectum and sphincter muscle revealed submucous tumor 3 (3
growth. DIAGNOSIS: Sarcoma recti (leiomyosarcoma). fl :5
CD
CD

Case 5. Ai44 year old man, who had previously been in perfect C0

health. During the years for 3 weeks each year he had

A
3

been exposed to herbicides while doing farm work in Denmark.

87

Employed in the oil-industry in 1954-68. During the summers of
1960-68 for about two weeks each year he had "rather carelessly"
handled phenoxy acids both with a sprayer and for spotting. Visited
a doctor in 1974 with complaint of increasing peripheral resis-
tance of palm and right underarm. DIAGNOSIS: Rhabdomyosarcoma.
Case 6. A 76 year old man, who had been healthy until 1972, when
he was operated on for a perforated ulcus ventriculi. Reported
some exposure to phenoxy acids for 4?5 summers in the 50's, at
which time he had cut and collected grass along roadsides, which
had been sprayed by the highway department. The patient also
reported that he had worked in forests which had been sprayed
with phenoxy acids. Visited a doctor for trouble with numbness
and pain in right arm. The palpation showed a 1 decimater long
tumor on the inside of the right upper arm. DIAGNOSIS: Myxo-

fibrosarcoma with SUSpected lipoblastic differentiation.

Case 7. A 67 year old forest worker with three earlier cases of
pneumonia and gastric anamnesis. The patient had worked in the
forests until 1970. According to his own account, had done cutting
and clearing from 1956 to the beginning of the 1960's within a
phenoxy-sprayed area. Indicated work in spraying for a few years

after this time. Since June 1969 he had noticed a peripheral

16396

Page 4

resistance in the left underarm. Visited a doctor in March
1970. DIAGEOSIS: Pelymorphocellular sarcoma, possibly a rhab?
domyosarcoma. I

In addition to these seven patients, among whom the exposure
to the phenoxy acids had been direct, at least in the first 
five cases, there are also a couple of patients both forestry
workers whose places of work in the 1950's and 1960's have been
occasionally located in proximity to sprayed areas. In one case,

harvesting berries after spraying was also reported. DIAGNOSIS:



Rhabdomyosarcoms and neurofibrosarcoma, respectively.

DISCUSS ION

The latent period for chemical induction of malignant, solid
tumors in man is generally assumed to be very long, on the average
ranging from 15 to 30 years. The latent period is possibly
dependent on dose (Heuper, Konway 1964). In the reported cases
the exposure occurred 10?20 years earlier. The contact had been
through the skin or through the respiratory system.

Of course we cannot draw any conclusions about the possible
causal connection between exposure to the phenoxy acids and
contaminants and the occurrence of malignant tumors, solely on
the basis of the reported cases. Exposure to the phenoxy acids
should be relatively common within the area of the three forest
provinces, from which the Umea clinic receives patients for
treatment. These cases very well could have been chance occur?
rences.

However, what is remarkable about these cases and should give
rise to continued investigations are the facts that these tumors
are of a rare type, that the exposure in all cases has been rather
massive, that the latent period agrees well with that assumed for

chemical carcinogens, and also that the distribution by sex fo

r;

malignant tumor at the Umea clinic strongly deviates.
from the national average. In a follow-up investigation of pos-
sible causal connections, extensive studies are needed, for example,
of epidemiology and industrial?medicine.

1s93?.



Maligna mesenkymala tumor
.?f'Qr fenomsyror en klinisk observatlon  

Siu fall med malign mesenkymal tumor iakttagna rid onkologiska kliniken i Ume?
?ren 1970?76 och med exposition for fenoxisn, or 10?20 ?r tillbaka beskrivs.
Expositionen har varit direkt och relatin massiv for fem av fallen. Latenstiden
st?mmer med den for kemisk karcinogenes antagna. Konsfordelningen i detta
material om totalt 87 patienter avviker Iran den genomsnittliga for riket med
manlig dominans. N?gra s?kra slutsatser om ett eventuellt kausalsamband kan
givetvis ej dras av dessa fall, utan for detta kr?ivs mer omfattande undersokning-

ar.

Fragan om fcnoxisyror (I ex hormoslyr)
cller i dessa ingaende fororeningar kan
utova skadeefl?ekter pa minniska och
djur har varit foremAI for en omfattande
dcbatt. Negon cancerogcnicitet hos man-
niska har hittills ej rapporterats.

I denna artikel beskrivs sju patienter
med malign mesenkymal tUmc'ir iakttagna
vid onkologiska Itlinilten i Umea Area
1970?76 och hos vilka exposition for fen-
oxisyror forelegat [0?20 {tr fore diagno.
sen av tumorsjukdomen. Dessa fall bevi-
sar naturliglvis inget 0m ett eventuellt
samband mellan sadnn cxpositioo och
malign tumor - for an bevisa eller ure-
slula ett s?dant samband kravs ing?ende
epidemiologiska undersokningar - men
det har 5nda motivcrat at! rappor-
tera iakttagelserna i kasuis?sk form.

Arm I970-76 har vid onkologiska kli-
niken i Umc? mottagits sammanlagt 87

tienler med malign mesenkymal tumor.

-1 dessa var 32 kvinnor (37 proc) och 55
min (63 proc). Av minnen bar 43 varit
yrkesverksamma; yrkesbeteckning sak-
nas dock i ett fall. Nio var skogsarbetarc
(20.9 proc). I'yra arbetade inom jord- och
skogsbruk (9.3 proc) och sex vid sagv'erk
eller massainduslri (14,0 proc).

Kasuistik

'3 Fall' I. 62-arig man. skogsarbetare
l'ram till pensionering pa grund av
dinrre' och smarter till viinster i buken.
RUkare'. V?rdad for (be I959. For ovrigt
\??sentligen frisk tidigare. I silt arbete
hade han 'besprutat med l'cnoxisyran
2.4.5-T knappt en vccka I963 och I964.
n-a veekor I965. en minad I966. en mi-
nad I967 samt Iva veckor I968. Sedan
auguin [976 iikande viirk i h?ger I?ossa
iliaca samt svullnad av hbger hen. Vid
laparotomi paiisades en Iumdri Iilla hiic-
kenet adherent tiil biiekenviiggen. PAD:
MedelhOgI dil'l'erentierat leiomyesarkom.

Fail 2. f7-arig skogsarbetare. icke r6-
kare. Hypertuni sedan I960. :Behandlad
mr postinf'eklids artrit I974. 133.1950. och
?U-talen under vardera tre ell? TII'yrasom-
tar och varje summar under are vcekor
hade han anviint en blandnii g-Iav 2.4-0
no? 2.4.5-T ?ckning och rzaindre ut-

kning fer mg. Sedan
nubusti I976 tillviixande tun iirwentralt.

LAKAHTIDNINGEN -VOLYM 74 - NR 33 - 9H

 

LENNART HARDELL iir underliikare vid
onkologiska klinikcn. regionsjukhusct. Umcd.

 

proximal! pa hoger Iarben. PAD: Mesen-
kymal tumor. troligen medelhogt diffe-
rentierat ?broserande liposarkom.

Fall 3. 49-arig man. verkstadsiigare.
Mangariga for ovrigt
frisk. I sin verkstadsloknl hade han aren
l96l?72 under icke besprutningssiisong.
ca Il minader Arligen. forwarat tusentals
liter av I?enoxisyror for skugsbolags riik-
ning. Forvaringen hade delvis skett i opp-
na hinkar. Patienten hade iiven handskats
med medlet. Uppgav standig Iukt pa ar?
beISpIatsen. Augusti I976 melena och

bulismiirtor. november :Inyo akuta buk-

symtom som foranledde Iaparotomi. Har-
vid pdvisades tumor i Iilla biiekenet med
engagemang av rekturn. urianI'Isa samt
oven/5x: till tunntarm. PAD: Mesenky-
mal tumor. snarast neuro?brosarI-tom.

Cl Fall 4. ISO-arig skogsarbetare. maulig-
hetsrokare. Ar I938 men
for ovrigt frisk. Aren 1961?66 hade han
ca 20 arbetsdagar per ar besprutat med
fcnosisyror. S?kte i auguin I97I for de-
I'ekationsbesviir. Palputoriskt i rektum in-
nanl'or s?nktern submukiist vh?sande tu-
mor. PAD: Sarcoma reCIi (leiomyosark-
cm).

Full 5. 44-;irig man sum titligare varit
helt I'risk. :Kren under \ardera
Ire veckurs tid hade han esponerat sig I'I'ir
herbicider under jurdbruksarhete i Dan-
mark. Anstiilld i uljeindustri ftren I954 -
68. Sommartid {Iren 1960?65 hade han
[v.1 veckor arligen hands-Rats "gilnsku
Ovarsamt? med l'enosisyrur bade med
spruta och i I?orm av I'ickning. Siikte I974
I'ijr resistens vulart hij'er un-
derarm. PAD: Rabdomyosarkom.

Fullti. 76-3rig man som varit ?ix?k till
I972 han opererades I'iir perl'nrerat ul-
cus ventriculi. Angav viss expenering 
I'enosisyrur under 4?5 somrar pa SO-ta-
let. vnrvitl Iran slaeit och tillvanitagit eras
Iiings viigrenar som vagverket 
Patienten uppgav dessutOm alt han arbc-

M- him

er och eXposition

(71mm 

K.

-

tat och vistatsi skog som beSprutms met!
fcnoxisyror. Sbkte I972 for domningar
Och viirk i hoger arm. Pulpatoriskt pdvi-
sades en decimeteriang tumor p5 insidan
av- hO'ger overarm. PADz'Myxofibrosarlc-
om med misst?nkt Iipoiglastisk differenti-
ering. i

Full 7. 67-Arig skogsarberare med Ire
tidigare pneumonicr samt gaSIritan-
amnes. Palienlen hnde arbetat i skogen
I970. Hyggesrensat enligt egen upp-
gil't fri'm I956 till bo'rjan av ISO-Inlet inom
fenoxisymbesprutade omrdden. Angav
arbete i anslutning till besprutning tiII eu
par Ar senare. Sedan juni I969 hade han
noterat en resistens pa v?nster underarm.
Sc'ikte for denna i mars I970. PAD: Poly-
mort?cclligt sarkorn. m?jligen cu rahdo-
myosarkom.

Utijvcr dessa sju patienter. hos vilka ex-
positionen fer I'enoxisyror har varit dirckt
?tminstone i de I'cm forsta fallen. Finns
ytterligare ett par patienter bagge
skogsarbetare - som halt sina arhetsplat-
ser p51 I950?60-Ialen tidvis forlagda i an-
slutning till besprutade Omraden. I 
can fallet har dessutom biirplockning ef-
Ier besprutning forekommit. PAD: Rab-
dOmyosarl-tom respektive neurulibrosark-
om.

Diskussion

Latenstiden for kemisk induklion av ma-
ligna. solida Ium?rer hos miinniska nose:
1' allmiinhet vara myeI-cet lang och i ge-
nomsnitt av storieksonlningen fir.
Lalenstitlcn iir miijiigen dusberuentle
Rummy I964). I de relaterade
I'allen exponeringen l0?20 iir anfII
i tiden. Knntakten har skett \ia hud och
inandninesurgnn.

Nligra slutsatser om ett eventuellt kau-
mellun exposition I'Lir {enmi-
syror och diiri ing?ende I'Oi'ureninear
samt maligna mesenkymitln tumiirer kan
dras frzin de relaleratle fal-
Ien. Espmitionen fi'rr I'enusisyror Iorde
vara relativt vanlig inum de Ire skogsliin
sum ulgor Umed-klinikens upptagnings-
omr?de ueh det kan myeket yiil ha varit
I'raga om cu slumpmassigt sammantrzi?
fande.

Vad mm dock ?r anmiirknings?irt och
kan mutivera fonszitta undersokningar or
det :ir I'rfiea om en tuni?rtyp.
at! expositionen i samtliga fall vul'il gam-

 

ka massiv. an Iatenstiden slimmer med?

den for kemisk kareinogenes 
samt aIt kiinsI'Iirdelningen I'iir maliena
mesenkymala tumiirer vid Umea-kliniken
stark! avviker {ran den genomsnittliga i

2753

bison?

0\
CD




Un?t

 

15398:

 

'l'uhud-I 



I nah-"If. tun; 

 z- Ammoniumnitratdamm och nitrit



Foretagsl?kare har ena sidan sv?rt att si?llva genomfora stora epidemiologiska
undersokningar inom sitt ansvarsomrz?zde men a andra sidan sv?rt att engagera
arbetsbelastade unlversitetsinstitutioner for sina problem. En liten expositions-
studie som den h?r redovisade har v?ckt debatt i press och radio. S?dana studier
horde, liksom debatten lnom lantbruket och dess framst?llare av kvavehandels-
g?dsel och liksom debatten om v?rt av overgodsling kv?vefororenade dricksvat-
ten, kunna form? de stora institutionerna att satsa an mera p? epidemiologiska

fr?gor om nitrat och nitrit.

I LKAazs gruvor i Malmberget anvander
man sedan l965 Sprangmedel bestaen-
de av 95 proc nitritfn'tt ammoniumnitrat
blandat med 5 proc dieSelolja. Det kallas
h?ir AN. pa andra st'zillen ANFO.

Som ett I'dtt luftblandat pulver sprutas
AN vid laddning med genom
slang in i form drivna borrhal. Hela tiden
men sarskilt niir dessa botjar bli f?drdig-
dammar det kraftigt pa personalen.
Tekniskt forehyggande itgarder mot
dammet har ej vant framgangsrika
och oro over t?nkbar explosionsrisk har
darfdr utbrett sig.

Nitrat har lange forekommit inom indu-
strin, sarskilt vid framsti?dlning av kviive?
och dessas senare hante-
ring i lantbruket. Man har hittills ansett
halson'sken av ammom?urnnitrat som
ringa. I v?ra dagar har dock tillkommit
ore for cancel-risk av nitrosaminer. bil-

 

 



riket. For en fortsatt utredning av ett
eventuellt kausalsamband kriivs ingaende
undersokningar av tex epidemiologisk
och arbetsmedicinsk karaltt'ar.

Litteratur

HUEPER C. CONWAY D: Chemical
Carcinogenesis and Cancers. Spring?eld.
Ill: Thomas l964

 

Malignant Mesenehymal Tumours

and Exposure to l?henoxy Acids-

A Clinical Observation

LENNART MD. Dept of Onco-
logy. Regional Hespilal. S-90l SS Umea

LAKARTIDNINGEN 74-. 2753-2754 1977

Seven cases of malignant tu-
mours observed at the dept of onculugy in
Utnea during the years l--76 and expo-
sure to phentny acid-u a pen'od ul? l?-Ill
years are described. ?as direct
and comparatively ma st in the of the
cases. The latent periu iriin agreement ?tth
that assumed for chermcil ilarcinogenesis. The
distribution between the series in this material.
of 37 patients in all. deviates from the national
average. with a dominaJtce if males. No deli-
nite conclusions conce ning a posmble causal
connection can he dr: 'n s?om these Cases:
this would call for ex ensivc investiga-
tions. 

2754

i

 

Arbetet utg?r Eran LKAB. Malmberget. dar

AB RAHAMSSON 5r rs-

ka. LElf KAGSTROM 
KARL CLAIR RENARD fore-
tagsliikare och KURT STENLUND 
Ingenjor.

anikeln redovisade analyser av methemo-
globin a: utl'orda vid kliniskt kemtska laborato-
riet. Centrallasarettet. Boden.

 

dade av vissa aminer och nitrit. Shdan
risk forutsiitter forutom narvaron av ni-
trat-nitrit aven forekornSt av sekundiira
aminer. Det 5r n?rvat'ande oklart om
aminer i gruvmiljo mer in i
vaijc annan miljo och vad de har.
Christoffer Rappe. organokemist i Umea
och nitrosaminspecialist. studcrar for
narvarandc denna fraga.
AN-sprangmedlet innehdller som till-
sats som anses minska
dammbildningen. Av dessa iir emcllertid
endast 5 proc sekundiira och dessutom
langkedjiga. C-
atomcr. Risken fOr att det bildas karcino-
gena nitrosaminer anses upphi?n?a niir
aminradikalen innehaller l0 C-atomer el-
ler fler.
Hiir redovisade studie iir inriktad pa
nitritsidan av problcmet.

Nitrat-omvandlingen
Nitrat kan omvandlas i vatten till nitrit.

?Omvandlingen gar lz'lngsamt men gynnas

av alkalisk miljo. tex i munh.?.lan. Den
ej ails i sura miljd men
oresorherat nitrat bor langre ned i turmen
ater kunna ombildas till nitnt genom bak-
ten'einverkan.

Otzinkhan om an ej siirskilt sannoltkt
fir det ej heller :ut nitrat pa lul'uagarnas
skulle kunnu till
nitrit och upptagas av kroppcn.

Nitrit hinds i blodet till hemoglobinet
och bildar methcmoglohin. Bmdningcn at-
no sammu siitt som mutsvarande med
kulmomnid en udirekt- reversihel hind-
ning. Delta till .sLillnad l'ran hindningen
med andra kviivct'dreningar som anilin.
som binds v-indirckt? och icke reversrhelt
och under samtidig flirstonng av hemo-
globinmolekylen. Den indirekta hiadning-
en tordc cj vura .dttuell i grovmiljun. Bada
bindningarna for till 
Mcthemoglobin anviinds som para-
meter l'or nitrat-nitrit-expositioncn.

 



Metnd for besti'tmning av smri miingder
methemuglol?rin iir sedan 1976 inl'drd vid
kliniskt kemiska laboratoriet. Centralla-
sarettet. Boden. Ovre norinalgr'ans for
methemoglobin it: 2 proc av det totala
hemoglobinet. 

Hygieniskt griinsvarde r5: dricksvatten
iir for:

nitrit 0.02 mgl?liter.

nitrat 30 mg/liter for barn och 50 mg!
liter for vuxna. .

Vid IOO mg nitrat per liter anses vatten
ou'anligt som livsmedel.

Expositionsm?tning

Stil'telsen Svensk' Detonikforskning har
pa Uppdrug av Stiftelsen Bergteknisk
forskning tagit upp riskproblemen vid
AN-(ANFO-lladdning. Preliminiira
NH. NOJ-matningar har gjorts pa damm.
dels pa ?ller och pa nitrat i vattent'as
med impinger. NH. NO, ?r stark: hygro-
skopiskt. Kalkyler av expositioner har
gjorts (Skarped I975).

har man hittills ej gjon nig-I
ra miitningar av den verkligu belastningen
pa miinniska i samband med luddning.?
Med darav utt'orde vi i Malm-
berget nattcn till den l2 maj ?976 en
liingc diskuterad expositionsmiitning par
tio laddare under jord (observerad gruppl
och nio borrurc under jord (referens-
grupp). Som parameter valde vi met-I
hemoglobin i blod.

Under aktuella gjordes:
kuntinuerliga ?ltermiitnlnear av total-
damm pa laddningsarhetsplatserna. Sam-
tidigt bestiimdes iiven halt av NO).
 NO: UCII  

En likurtao utl'ordes nut--
ten till den 19 nktober I976 pa elva lad-?
dare och elw. borrarc. Serie Ill-12.

Storleken av gruppernu begransades av
resurserna pa Sodens centrallahorntorium
och vara cgua tallgang-E
:Ir. 

SamJI-ga iaddare och borrare hade
samlats I'dr instruktion dagen lore mat-
ningerna. Samarbcte med fackforeningen
upprattades. l-lan Rum Overens om all.
inte skulle anvandas av
vid (an-
vlinds l'iir (lvn'gt siillan pa grund av att
ammoniumnitrat kittar ihop ?ltret) och
:ztt gronsalter. siirskilt dd Spenat. ej skulle
inga i (Nitrat. se nedan!)

Elter skil'tslutet lugs kl 20.35-23.05
grover i or under jord. -
l'rovcrna. Ill ml \?C?biUd i vakuum-hepa-=
rinri?ir pluccrades Omedclhart i 
och snahhehandlades sedan.

scrie 21?42 hlev analysen l?otsenad
ett dan genom ett tekniskt miswde inum
Llimskt-kemiska lahoratonet i Linden.

Samtidigt logs prover pa nattpersonal

och patienter 

sarett.

 

IVOLYM 74 -NR 13 - i977

 

?a a 2445
ECOLOGICAL BULLETINS No 27

5

Chlorinated Phenoxy Acids and 
I I 0
?Their Dioxms 
Mode-of Action, Health Risks and 
Environmental Effects 
eps-

Cc:

C. Ramel led.)

1978. Ecological Bulletins. Swedish Natural Science Research Council.

ISBN 91-546?0234-3

Production and distribution: NFR Editorial Service
LFIALLF 209 T7 002

Printed in Sweden by Berlings. Lund 1978

Copies of the bulletins can be ordered from the Editorial
Service of NFR. Box 23i36. S-l04 35 Stockholm. Sweden. The
Editorial Service. which provides ?nancial and practical assistance
for the publishing of scienti?c works. is responsible for the distri-
bution of all NFR publication. 

Report from a conference arranged by the
. Royal Swedish Academy of Sciences
. Stockholm, Sweden, 7?9 February 1977

micereef. asst?- em

 



. . abb- . WE . +124 1,5, y" 35LIST OF CONTENTS

Editor?s preface
C. Ramei

CONCLUSIONS AND RECOMMENDATIONS

CHEMISTRY-
Summary

Chemical background of the phenoxy acids and dioxins
C. Rappe

Analysis of in the Seveso area

P. Adamoli, E. AngeIi, G. Bandi, A. Berroloni. E. Bianc/zi, L. Bumforti.
I. Camoni. F. Carmbeni, G. Colli. M. Colombo, C. Corradi. L. De Angelis.
G. De Felice. A. Di Domenico, A. Di Muc?cz'o. G. Elli. R. Fanelli.
M. Firtipaldi, A. Frigero, G. Galli. P. Grassi, R. Gualdi, G. Invernizzi.
A. Jemma. L. Luciano, L. Manure. A. t?bhlrine?n. F. Merit] S. i?v'icosia,
F. Rizzello, C. Rossi, G. Rossi. G. Salvatore, A. Sampolo, G. P. Schmidt.
F. Taggi, G. Tebaldi, E. Zaino G. Zapponi.

The problem: a review
D. Firestone

Analysis of in environmental samples
J.D. McKinney ?ne-HS

decontamination

F. Poc'chiari 

Con?rmatory tests for pesticide residue analysis
G. Widmark

Photochemical reactions of phenoxy acids and dioxins

B. :lkermark.
1 woe-:2: was-.5   
if?) (Eff-A as 9 11'. IS. ta?
  r) K. 515.?. 6'5? 

 

a (1.3V.
5" 1? 

to. -.
. -. .
en" - -

5..


i

nuburn-v.55

I .



.5:

.-






 

{gases 



PLANT PHYSIOLOGY

Summary

The herbicidal effects of phenoxy compounds
B. fiberg L. Eliasson

TOXICOLOGY
Summary

The phenoxy acid problem in Sweden
J. 

Phenoxy acids: effects and fate in mammals
P. J. Gehring cf: J. E. Berso

Toxicity of 
J. A. Macre

and enzyme induction
A. Poiand E. Glover

Phenoxyacetic acids: sublethal effects
P.-0. Sjiid?n U. Sb'derberg

    
      

 

83 ECOLO
85 Summary
86 2 Chemical
modes of.
U..Gransr

108
Phenoxyat
considerat
122 B. 
c" Effects of
134 In Sweden
T. [11881th
?45 Bioaccum
A. R. 
149

Effects of

L. Torsu-
effects and mechanisms [65 Hmhmim
?llh 2.3..
A. H. H.
List of pr
List of 0 
GENETICS List of 
Summary 17?! ?3 
Genetic effects of phenoxyacetic acids in animals 199.
C. Ramel
Genetic effects of chlorophenoxyalkanoic acids on plants 186
L. Ehrenberg
Mutagenicity tests of 2,4-dichlorophenoxyacetic acid and 2,4,5-trichlo- 190
phenoxyacetic acid in genetically stable and unstable strains of Drosop/u'la
melanogaster 
B. Rasnmson cf: H. Svahlin
Genetic effects of phenoxy acids on microorganisms 193
G. Zetterberg
r?s-a-    

r?

 

 

-

. -. 
1-ng rep. .
- ?na?am  

:3 l_ ECOLOGY AND ECONOMY 205

:5 I Summary 207

5 Chemical weed control in the tropics 210
G. E. Blackmail cl; J. D. Fryer .

The use of phenoxy acid herbicides in Swedish forestry: amounts, types and ??19
modes of application
U. 5L M. Bt'irring

The use of phenoxyacetic acid herbicides in Swedish agriculture 231

8 B. 
Phenoxyacetic acid herbicides as a tool in forest management: silvicultural 235

1 considerations

B. 
Effects of the silvicultural use of phenoxy acid herbicides on forest vegetation 240

4 in Sweden
T. Ingelb'g


cc. to

5 Bioaccumulation of 255 
A. R. Isensee 

.763 ?y
Effects of phenoxyacetic aetd herbmdes on organisms 
Torstensson

5 Ecological considerations regarding massive environmental contamination 285
with 
A. H. Waring
List of participants I 295
List of observers . 297
List of additional authors 299

6

0

3

7

 

?sitter-tr": its-1' Me 
Jud-n1 if 
52565nil".- gri?cs?v  

   

  new  



i



A . . 3"



 
 
  

u?nu- . -.- -.
TN ?hr? . 
- - Ir.' 

Ramcl. C. led.) I978.
Chlorinated Phenosy Acids and Their Dioxins
Ecol. Bull. (Stockholm) 27:13?16.

CHLORINATED PHENOXY ACIDS AND THEIR DIOXINS:
CONCLUSIONS AND RECOMMENDATIONS

Conclusions

The phenoxy acid herbicides are not formed naturally in the environment. There
is no evidence that dioxins are formed from phenoxy acid herbicides in the en-
vironment. The only dioxin ofenvironmental concern with respect to the phenoxy
acid herbicides is the dioxin
found in the herbicides, acid and 2.4.5-
trichlorophenoxypropionic acid TCDD is found neither in 2.4-di-
chlorophenoxyacetic acid (2.4-D) nor in acid
(MCPA).

Levels commonly encountered in soil after normal applications of phenoxy acid
herbicides are parts per million (ppm); due to heterogeneity of soils and
irregularity of application. these levels could be somewhat higher or lower
locally within sprayed areas. The half-life of the phenoxy acid herbicides is of the
order of 3-4 weeks; that of is up to 10 weeks. The rate of degradation
generally increases with increasing microbiological activity and decreases at
lower pH. especially below pH 4. Repeated applications of 2.4-1) and MCPA
result in somewhat faster microbiological degradation.

The level of phenoxy acid herbicides in the green parts of target vegetation
shortly after normal Spraying ppm. After 10 weeks, this level decreases
to 20?50 ppm. During the cold winter period this value seems in some cases to be
nearly constant, although in other cases it decreases. lnjecred phenoxy acid
herbicides may persist for 4?6 years within the woody parts of vegetation.
however, the ecological consequences of such persistence in wood are probably
not very great.

Photolysis is the primary mode of degradation of TCDD on environmental
surfaces; microbiological degradation occurs slowly in soil. Its hal?ife in soil has
been estimated in the United States to be l?l.5 years (on the basis of studies in
which high levels of dioxin-contaminated phenoxy acid herbicides were used).

Wind-drift of Sprayed phenoxy acid herbicides may reach non-target areas.
To avoid such risks. strict regulations with regard to weather conditions during
spraying and with regard to spraying techniques are required.

There appears to be virtually no downward movement of TCDD in the soil:
it can be estimated that no more than 1% is transported from the site of applica-
tion annually. Movement in the air would occur largely via airborne particles.
Vegetation takes up very little or no TCDD from soil.

l3

  

 

      

*1 NW
gig-1&1:  .  
I. lg}

hank15. ~53 1-2. . 5-

.9 
333??: I: ?wi?h? 
as 1?5? ii; arm-matT?r vs" 9?


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h?

  
  
 

 

   

when


..
.

 
 
 
 
 

. 

   

 
- . . 
. 

    
  

   

1.
343:1



.3-

    
 
  

?it?

    
 

1.1.--
avI.le  -

 1
13;. .

There is no evidence ofbioaccumulation of phenoxy acids; bioaccumulation of 
TCDD resulting from normal use of 2.4.5-T is insigni?cant.

The fate of phenoxy acids in plant tissues. changes in plant composition
induced by these compounds and their mechanism of toxic action are understood
only imperfectly. It can be concluded that further studies in this ?eld would be of
importance for understanding the response of cr0p plants to phenoxy acids and
for the development of more selective and efficient methods of herbicide usage.
Such studies would also provide important information about aspects of herbi-
cide toxicity at the cellular level that may be relevant to their toxicity in animals.

Possible levels of exposure to phenoxy acids and/or dioxins vary among
different populations. such as: employees in factories where phenoxy acid herbi-
cides and/or trichlorophenol are made; persons who prepare and apply the
herbicides. and members of the general public who enter sprayed areas andlor
consume treated vegetation. People representing potential risk groups. pregnant
women and individuals with chronic liver disease, should be excluded from
activities involving high exposures. such as manufacturing and occupational
spraying.

Only very low levels of phenoxy acid herbicides are normally found in food-
stuffs. Levels in Swedish wildlife. ?sh and waters are also low. The residue levels
in woodland berries from recently sprayed areas might be higher, of the order
of 10 ppm. A daily intake of as much as 1 kg of such berries would correspond to
a dose of0.2 mg kg'I clay'1 of phenoxy acids for a person weighing 50 kg. This
level probably represents the same degree of exposure as that realized by herbi-
cide Sprayers. Calculations based on extreme conditions (high exposure from food
and lowest dose of 2.4.5-T that caused teratogenic effects in mice) still provide
a safety factOr of at least 100.

If the levels of TCDD in phenoxy acid herbicides remain below 0.l ppm.
considerations of health risks associated with these products should be based 
primarily on the potential toxicity of the phenoxy acids per se and not on that of
its cuntaminants. .

In evaluating the risk associated with the presence of TCDD in phenoxy acid
herbicides. potential exposure to compounds that have similar toxic effects.
other halogenated dibenzodioxins. dibenzofurans, naphthalenes and azoxy-
benzenes, should also be considered.

Studies to assess the tumorigenicity of phenoxy acids and TCDD provide no 
de?ntive indication that these agents are or are not carcinogenic. Additional,
more extensive evaluations are underway.

The only statistically signi?cant demonstration that phenoxy acids induce
mutations transmissible to the was obtained in the recessive lethal test
in Drosophila melanogaster. Work on microorganisms sustains the conclusion
that the phenoxy acids are mutagenic under the test conditions used. Two in-
vestigations on 2.4.5-T and one on 2.4-D. both using pure acids. showed an in-
creased number of recessive lethals in Drosogiltila. From a tentative comparison
of the radiation dose that would give a similar effect, the mutagenic effects of
and were concluded to be weak. 

Somatic mutations have been observed in Drosopliila. and chromosomal
aberrations have been shown to be induced in plants and mammals. The relevance
of these latter ?ndings to a risk of mutation from phenoxy acids (associated or not





1-1

164051

 



-- is?. "61.x; .  
  genome?r?ges?ft?gm a?  1535.  3 .ggu . . Fq?zn?. I (Elf-eta-art?s-rui- sth??fs? . .1. teas 

.- 



- ,with heritable damage) requires further studies. Observation in one study of
a possible role of the emulsi?er and/or solvent show the necessity of including
these constituents of phenoxy acid formulations in future studies.

In view of positive mutation in bacteria and ?ndings of chromosomal
aberrations in one mammal and in one plant species with TCDD, it must be con-
cluded that this compound is mutagenic. The fact that other tests ofa similar kind
were negative may have many explanations. such as problems in keeping the

compound in solution. Such dif?culties make it impossible at present to quantify

genetic risks from TCDD.

With regard to behavioral effects of the phenoxy acids in mammals, it was
concluded that further studies of dose-response relationships are necessary
before it would be possible to make an evaluation of similar effects, if any, of the
doses of phenoxy acids to which humans could be expected to be exposed in
connection with normal herbicide usage.

No counterpart to the physiological mechanism involved in the major herbicidal
effeCts of phenoxy acids in plants is known to exist in animals. In plants. high
concentrations of phenoxy acid compounds cause less speci?c effects, the
uncoupling of oxidative phosphorylation; this effect appears to be weak in
mammalian organisms.

Recommendations

l.The stability and bioaccumulation of TCDD should be studied in realistic
situations. under normal ?eld conditions. in various types of climates and
soils. 

2. Risks associated with exposure to TCDD could be greatly reduced if manu-
facturers would utilize processes, currently available. that minimize the
formation of TCDD from trichlorophenol. Manufacmrers should also use
techniques that minimize potential contamination of the surrounding environ-
ment by TCDD in the event of indusnial accidents. In addition, striCt precau-
tions should be taken to ensure safe diSposal of TCDD-containing residues.

3. Failure of manufacmrers to utilize techniques that minimize TCDD formation
or contamination should lead to a re-evaluation of the feasibility ofcontinued
use of trichlorophenol or of products derived from it. such as the phenoxy
acid herbicides, and 

4. The use of other processes that eliminate formation as a
contaminant should be investigated.

5. At present. the mechanism for the toxicity of TCDD is unknown. A further
understanding of how TCDD and related compounds act is necessary.

6. Further sutdies of the mutagenic aetivity of the phenoxy acids are important
and should comprise all constituents of formulations and their impurities.
In order to make quantitative estimates of risk to man. it is essential that th
mechanisms for the genetic effects observed be clarified. - 

7. Studies of muragenicity should also aim at safe-guarding plant genotypes.

8. It is recommended that epidemiological investigations be made of human
groups exposed to phenoxy acid herbicides and dioxins and that these com-
prise all effects that have a possible genetic mechanism.

9. In forest use of herbicides, it is recommended that consideration be taken

 
 
  



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of non-target plants within target areas, eSpecialIy those of recreational and
scienti?c interest. Information about the effects of herbicides on non-target

- ?ora is insuf?cient. and further research within this ?eld is recommended.
10. It is recommended that Special consideration be given to the ecological

importance of ?eld borders and other ecotones in any spraying program. 
II. In choosing herbicides. their selectivity for intended target Species and other 
species within target areas should always be considered. 
12. The ecological importance of habitat. including both food and cover. to C3
vertebrate and other fauna was recognized; it is recommended that this factor 

be taken into consideration in the use of herbicides and in other manipulations 

of habitat. Further research on the possible signi?cance of alterations to wild? 
life habitats is recommended. Both vertebrates and invertebrates should be C)
considered. I 


l3.The conference stressed the importance of establishing and maintaining
reference areas representative of major ecological types. With reSpect to
herbicide use, it is recommended that areas are set up in which herbicides
would not be employed and in which other management Operations are strictly

controlled.

 

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MIDLAND. MICHIGAN 48640

9008 Building
May 21, 1971}

CONFIDENNAL - su

8
ED. ML 44 JECT TO INJUNCTION

AGREEMENT 9-79.

 

J. H. Davidson, Manager 0
2,14,5?1" Project 0.
9008 Building i?
CD

cc: E. H. Blair 1.1. M. Theis '3
M. C. Feigelson M. J. Traynor be

G. E. M. R. Wessel 50

D. D. McCollister C. S. Williams 

SILVEX AND ERBON - REGISTRATION AND TOXICOLOGY

On May 16, EPA published a notice of intent to broaden the 
hearing to include all registered uses of herbicides derived from
2,h,S?trichlorophenol, including silvex and erbon (Federal Register i2,
No. 96, p. 17h66). Dow is the principal manufacturer of silvex and
the only manufacturer of erbon. Current Dow registrations for silvex
are:

Low Volatile Brush and Weed Herbicide-(EPA Reg. No. h6h-162)
Silvex Acid for Manufacturing Use Only (EPA Reg. No. h6h-h55)
Silvex Butoxypropyl Esters for Manufacturing Use Only (h?h?hSl)
Silvex Isooctyl Esters for Manufacturing Use Only (h6h?h36)

Registration for other Dow silvex products was cancelled in October
1966 when Dow indicated they would not renew their registrations a
required every five years. -

GARLONQ Grass and Weed Killer (hGh-ElT)
KUROSALQ SL Aquatic Herbicide (14614-27'?
KUROSADD Aquatic Herbicide (h6h?276)
COLORSETO for control of preharvest drop in apples (h6h?267).

Other products similar to include:

SILVI-RHAPO (formerly Hercules, now Transvaal)
(Amchem) -
Silvex (Diamond-Shamrock)

Silvex ML (Chipman Division, Rhodia, Inc.)

Silvex LV (Thompson-Hayward).

Salt formulations used for aquatic purposes are registered by Pennwalt,
as mixtures with Dilute formulations of

amine salts are used as growth regulators in fruit crops gammy:
Arcadian STA-SET EPA Reg. No. 218-h92, Allied 5 3
Chemical Corporation}. A large portion of the estimated 5

AN OPERATING UNIT OF THE DOW CHEMICAL COMPANY y?all ms

164GB

J. H. Davidson -2- May 21,.19Th

1.5 million pounds of silvex produced annually is used in the home and
garden market Chevron). 

With regard to erbon, we have three registered products:

ERBON Non Selective Herbicide (formerly h6h?172)
Weed and Grass Killer (h6h?255)
ERBON for Manufacturing Use Only (h6h-335)-

The latter contains h2% erbon and 15% related chemicals which are not
designated but are claimed as active ingredients. It is sold to many
formulators for registration under their labels for home and garden use.
Examples include: 

Greenfield Contact Killer (1&71-33. Elanco or Eli Lilly)
Best Liquid Edger Occidental)
Best Ded n' Gon or All-Out Weed Killer Occidental)
Four Seasons Edge or Trim Grass and Weed Killer (7393-UE, Wonder)
Ferti-Lome New Improved Perms Trim (Voluntary Purchasing Groups)
Weed and Grass Killer No. and No. 2 (hOT-xx, Imperial)
Green Light Liquid Lawn Edger (869?61)
Sears Liquid Edger (539?25h)
Uses, rates of application, toxicology, residues, and status of regis-
tration are discussed individually below for silvex and erbon. Additional
details can be provided as necessary.

Silvex is the common name for acid.
(Note that it is a mixture of and forms of the chemical due to the
presence of an asymmetric carbon on the side chain.) According to the
current EPA Summary of Registered Agricultural Pesticide Chemical Uses,
it is registered for use as an herbicide at rates up to lb/A in pasture
and rangeland, 1.5 lb/A in rice, 5 lb/A in sugarcane (Florida, Louisiana
and Hawaii), and he lb/A in lakes and ponds. It is also registered for
use as a growth regulator applied postharvest at 7.5 on pear trees
and preharvest at 20 in apples and prunes, with a maximum rate equi?
Valent to 60 grams per acre. Uses for control of brush in industrial
and other non?cropland areas include rates up to 16 lb/A as approved by
EPA in October 1973 for contains 69.2% propylene glycol
butyl ether esters of silvex, or h5.8% silvex acid equivalent, which
amounts to lb a.e. per gallon.

We have considerable toxicology data for silvex, including reports on
subacute 90-day studies with the sodium salt and the PGBE ester in rats
and with the potassium salt in dogs. We also have chronic 2?year studies
with the potassium salt in rats and dogs, and recent teratogenic studies
with the acid and the PGBE esters in rats. The results of these are
summarized in the attached excerpts from Section of Pesticide Petition
No. 8F06TS. EPA has indicated that they are satisfied with the studies
but that a multigeneration reproduction study would be needed.for toler?
ances greater than 0.1 (negligible residue) in food crops.

leias



J. H. Davidson May 21, 1971.

All registered uses for silvex in food crops are covered by PP 8F0675
first submitted in December 1967 by the Industry Task Force fOr Phenoxy
Herbicide Tolerances (formally disbanded in April l97h). Amendments to
the petition were submitted by Dow in September 1968, December 1970,
October 1971 and January 1973, for a total of nine volumes to date. The
petition is currently being held in abeyance until 11/1/7h pending com-
pletion of.a study on persistence of silvex in soil and of another feeding
study in cattle.

The residue pattern for silvex in milk and meat resembles that for other
phenoxy herbicides in some cases but not in others. For example, residues
were not detected in milk of cows fed silvex, 2,h-D or MCPA until the
level in total diet was raised to 1000 ppm, but residues of 2,h,5-T and/
or 2,h,5?trichlorophenol occurred in milk when the cows were given as
little as 100 2,h,5-T in the diet. Residues in meat tissues, parti-
cularly liver and kidney, were much higher for silvex than for 2,h-D and
MCPA and were substantially higher than for 2,h,5-T at levels of 300 to
2000 in the diet of cattle and sheep. However, residues of the
2,h,S-trichlorophenol metabolite were low or non-detectable in milk and
meat of animals fed silvex in contrast to relatively high levels of the
phenol in milk, liver and kidney of animals fed Additional
feeding studies are currently being conducted at reasonable levels of
300 in the diet, to determine the preslaughter intervals required

to reduce residues of silvex (or of 2,h,5-T plus trichlorophenol) to
below the 0.1 negligible residue level in liver and kidney. We have
proposed limiting the rate of use to 2 lb/A in pasture and rangeland,
with no cutting of hay during the season of application of silvex or

2: 

Interim tolerances were established in late 1972 for many pesticides
formerly registered on a no-residue basis in food crops. Included was
silvex at 0.1 in or on apples, plums (prunes), rice and sugarcane.
(Federal Register No. 169, p. 1775, August 30, 1972). However, a
subsequent order proposing additional interim tolerances, including

2,h-D and MCPA in grass, stated that interim tolerances could not be
established for 2,h,5-T in any crop, nor for silvex in or on grasses
(pasture and rangeland), grass hay, and in water. The reason given was
because the data in petitions and otherwise available data were Judged
inadequate to rule out the likelihood of injury to consumers from residues
from these uses (Federal Register 313 No. 178, p. 18565-18566, September
13, 1972). Dow appealed this for silvex in grass but the appeal was
denied. In February 197k, we renewed the request for an interim tolerance
for silvex in grass but EPA replied on April 5, 197h that data from the
additional feeding study were needed before they could finalize the usage
on grass, i.e. application rates, withdrawal times, etc.; hence they

could not establish interim or permanent tolerances for grass at this
time.

Use of silvax as a growth regulator in pears was cancelled by EPA in

March 1972 when they denied our request for exemption from the need

for tolerances. We had hoped they would consider it a non?food use 
since the 7.5 spray is applied to the trees after harvest of the pears,

18410-

I Q7050 4A a 

   
- ?r a 

H. Davidson May 21, 1971:

i.e. a year before harvest.of the next crop. However, EPA ruled that
this use would reasonably be expected to result in small residues in or
on harvested food or feed. Subsequently, we provided all available
data on silvex in pears for submission by Inter-Regional Project No. 
on behalf of the State of Washington. Their Pesticide Petition No. -
3E1339 was deemed adequate by EPA and a permanent tolerance of 0.05 
was established for silvex in pears (Federal Register gg, No. 165, p.
2289h, August 27, 1973, 5180.3h0 of the Code of Federal Regulations).

Uses of silvex as an aquatic herbicide are covered by Pesticide Petition

No. lElOl2 submitted by the U.S. Corps of Engineers in conjunction with

the Bureau of Reclamation, USDI. However, EPA has repeatedly found their
petition inadequate. We have supplied them with information for incor?
poration in their petition and have authorized EPA to consult our petition
8F0675 on their behalf. EPA has raised questions about residues of meta-
bolites in fish, and the possibility that TCDD might be formed by cooking

of fish containing residues of trichlorophenol. Their last amendment

was submitted in November 1973 and their cover letter included the statement:

"We understand that a hearing will be held by the Environmental
Protection Agency in April l9Th. If this hearing includes the

aquatic use of silvax, we would like to defend our position on

the basis of Technical Report No. 5 Aquatic Use Patterns for

Silvexf" 

This report dated October 1973 is 1/2 inch thick.? A copy was sent to

M. Leng by Dr. E. 0. Gangstad of the Corps of Engineers as part of the
inter-communication between Dow and the petitioners for tolerances in
water and fish. The first sentence of the Foreword states that an announce-
ment by Dr. Lee A. DuBridge in October 1969 listed "actions being taken
by governmental agencies to restrict the use of the herbicide 2,h,5?
trichlorophenoxyacetic acid (silvex)". Apparently they don't know the
difference between the various compounds since they also reported finding
as a metabolite of silvex in birds. (Dick Otten of Amchem is

a member of the Registration Work Group of the Interagency ad hoc
Committee on Use of Herbicides in Aquatic Sites.) 

Erbon is the common name for the herbicide chemical 
phenoxy)ethyl 2,2?dichloropropionate. It is the alcohol form of 
combined as an ester with Dalapon. It degrades slowly in soil, presumably
to 2,h,5-T and Dalapon which kill broadleaf weeds and grasses, respect-
ively. When used at high rates, it eradicates all vegetation.

Dow's original product was called Non Selective Herbicide, which
was reregistered in 1971 as ERBON Non-Selective Herbicide under the
same registration number (h6h-172) after we sold the trademark 
for other purposes. It contains 30.5% erbon and 10.5% related compounds
for a total of lb/gal. It is recommended for use in industrial areas
'such as storage and parking areas, tank farms, rail sidings, substations,
outdoor theaters, drainage ditch banks, fence lines, pole yards, lumber
yards, elevators, equipment storage.areas, around farm sheds, and in
non?crOp agricultural land. It is usually applied in late May or early
June in northern arcas or in early winter or mid-winter in California.
Recommended-rates are 3 to quarts per thousand square feet or 30 to
ho gallons per acre. This amounts to a maximum of 160 lb/A including
ho lb/A "related products". 3

15411

J. H. Davidson -5- May 21. 197h

Dow also has a private label product called Weed and Grass Killer
(EPA Reg. No. This contains 9% erbon and 3% related compounds
for a total of lb/A. It is recommended for use in driveways, walkways,
parking areas, under fences, around garages and other buildings, and in
similar areas where vegetation is not wanted, as well as for edging
various areas. The rates are described water applied
to 250 sq-ft, or to 100 sq ft when extended control is desired. This
amounts to a maximum of l/h pound per 100 sq ft or about 110 lb/A
including about 30 lb/A "related products". Presumably, other products
formulated from ERBON (h2% erbon, 15% related products) are recommended
for use at similar rates. -



Acute toxicity data on erbon and ERBON indicate an acute oral LDso of

to 2 g/kg. Precautionary labeling indicates it may cause skin irritation
and irritation. Toxicity studies have been conducted in fish (2/28/63)
and in birds (submitted to EPA on 9/26/73). According to Warren Crummett,

analyses for TCDD are complicated by the high content of other materials

in our technical erbonHl?qu  

?5?nifauwki-

Marguerite L. Leng
Government Registration
Health Environmental Research

r"

 

enc:3

P. Sr See attached article on ERBON plus in the latest
issue-of the Dow publication "Industrial Vegetation Management"
(Vol. 6, No. l, l9Th) by Juell Johnson of the South Dakota
:Department of Highways. Newly approved uses of are listed
in an excerpt from the same magazine as an addendum to an Update
on the Status of 2,h,5-T by L. Southwick.

16412

 





rr cruel: Silvex - Section 6.1, October 1971
Toxicology

A summary of known information on the toxicology of silvex

was included in the December 1967 submission, and was repeated
as Sections C.0.l and C.0.2 in the amendment submitted in-'
December 1970. Additional reports on-toxicity studies in

r. 

cattle and sheep were included in Section C.l in December 1970.





Data presented in these submissions Show that the oral LD50
of silvex and its various formulations is in the range of 500
to 1000 mg/kg of bddy weight for various species of animals.

A no-effect level of 7 mg silvex per kilogram per day was

found in 90-day feeding studies with the sodium salt and the
propylene glycol butyl ether ester in rats, and with the potas-
sium salt in dogs. The no-effect level in Z?year feeding

studies was essentially 2.6 mg/kg/day in both species.

No significant adverse effect was produced in livestock given
salts and esters of silvex orally at levels of 25 or 50 mg/kg/da
However, 100 mg/kg/day was lethal to cattle and sheep after

29 and 11 daily doses, respectively, and 250 mg/kg/day produced
death in cattle after five doses. Chickens showed reduced
weight gain after ten doses of 100 or 250 mg/kg/day.

The lethal dose of the butoxyethanol ester_of silvex was >9000
mg/kg of body weight for pheasants and young Bobwhite quail.
Mortality was produced in 50% of young Bobwhite quail given
the ester at 5000 in the diet for less than 10 days, in
young pheasants_at 5000 for less than 100 days, and in
adult Mallard ducks at 2500 for less than 100 days.

16413

5.

{we

Summary of Toxicological Data on Silvex in Rats and Dogs

 

 

 

Report Formulation Test Duration No-effect level mg/kg/day
and date used animal of test given in report silver equiv.
Bio-Test Labs Dowco 171* Rats 90 days 10 mg/kg/day 6.8
6/19/62 (PGBE ester)
Dow Chemical Sodium Rats 90 days 0.01% in diet 5-10
10/26/54 salt (calculated)
Dow Chemical Kurosal SL Dogs 89 days ?0.03% in diet 7
10/30/63 (K salt) - (based on daily
food intake)
Dow Chemical Kurosal SL Rats 2 years 100 in diet 2.6
3/17/65 (K salt)
Dow Chemical Kurosal SL Dogs 2 years 190 in diet 
4/8/55 (K salt) I
*?ropylene glycol butyl ether esters of silvex (67.5% silvex acid equivalent). KURCNG weed

and brush killer contains approximately 68% PGBE ester of silvex (45% silvex acid equivalent).

nistopathological effect was noted in male dogs but not in female dogs 2.6 /Ig,cay silver equivalent after two years of continuous feeding.

Prepared by Marguerite Leng, Agricultural Department, The Dow Chemical Company, December 29, 1966

Revised September 10, 1970



a.  

Section C.2.0 Summary of Teratology Studies with

Silver in Rats. January 1073.

 

Numerous terat010gy studies were conducted with phenoxy
herbicides by various government and industrial groups

following disclosure on October 29, 1969, by Dr. Lee A.



DuBridge, Science Adviser to the President, that studies
cenducted in 1965?1968 by Bionetics Laboratory in Falls
Church, Virginia, indicated teratogenic responses were
produced in mice and rats given high doses of 
The significance of these and other studies conducted

at exaggerated levels by non-representative routes of
administration is still under discussion by experts in

the field of teratology.

As part of this pregram, studies were conducted by The

Dow Chemical Company on silvex and its propylene glycol
butyl ether (PGBE) esters by oral administration in rats.
The two studies were reported in Sections D.2.l and D.2.2
of this amendment to PP SFOGTS for silvex. Groups of

- Sprague Dawley rats were given silvex at doses of 0, 50, 75
or 100 mg/kg/day on days 6 through 15 of gestation, or from
day 6 of gestation through lactation. The test compound
was administered orally Once daily as a corn oil suspension.
Additional groups were given Silvex at 25 to 150 mg/kg/day
or its PGBE ester at 35 to 200 mg/kg/dayJ on days 6-15,

7-9 or 9-11 of gestation as described in the reports.
Certain groups were utilized for teratologic studies while

others were used.for postnatal studies.
The following parameters, where applicable, were examined

in these studies with silvex and its PGBE esters: clinical

observations; body weights; food consumption; :ross visceral

16415.

 L-J l_l4 





examination of dams; position of fetuses in utere; numbers
of live fetuses, resorptions and corpora lutea; individual
pup weights (days 0, 5, 21) and sex; external skeletal and

Visceral ?xaminations on toratology litters; histologic i;
examination of selected tissues; examination'of weanling 
rats to determine degree of skeletal ossification. f?C

CL

Conclusion

Ne overt clinical signs of toxicity,.major visceral or
skeletal anomalies or undesirable effects on other repro-
duetive parameters resulted from treatment with silvex at
25 mg/kg/day or silvex PGBE esters at 35 mg/kg/day (equiv~}
alent to 23 mg of silvex per kilogram of body weight per

da?<on days 6 through l5 of gestation Doses of 50 mg/kg/

'day resulted only in fetal toxicity as reflected by decreased

fetal weight and minor skeletal variations, but no majon
abnormalities were observed at this dosage level.? Maternal
and fetal toxicity and fetal anatomical alterations were
observed at levels of 75 mg/kg/day or more for silvex and
at 100 mg/kg/day for silvex PGBE esters. Maternal death
occurred when silvex was administered at 100 mg/kg/day for

10 days during gestation.

I?e


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DICI. 5.3. 50.3.; Lit-{Ill   

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s.

 

MIDLAND, 435m 
9008 Building Vic
August 10, 1976 v- a;

M. w. Sauerhoff
Health Environmental Research
1803 Building

 

P. J. Gehring, 1803
cc: E. H. Blair, 2020 H. L. Gordon, 2030
G. E. Blau, 1707 J. E. LeBeau, 1803 
M. 8. Chenoweth, 607 D. D. McCollister/Regi Section 
J. H. Davidson, 9001 B. A. Schwetz, 1803

REPORT ON SILVEX STUDY IN HUMANS

Thank you for sending me a copy of your report on Fate of Silvex Following
Oral Administration to Humans, dated May 21, 1976. It represents a
tremendous amount of work and should be very helpful in resolving some of
our problems with this herbicide. However, as we discussed by telephone,
a somewhat different treatment of the data could provide even more useful
information on rate of excretion of silvex by humans, for comparison with
and 

First of all, I believe the half?life should be based on total excretion

of silvex plus silvex conjugate in urine and feces rather than on only free
silvex in urine. This w0u1d take care of variations in proportion of
silvex in free and conjugated form in urine of the various subjects,
ranging from a ratio of 5.19 for subject 1 to 0.66 for subject A (Page 12).
Would it be possible to have Gary Blau run the computer analysis of data
for silvex plus silvex conjugate for successive 24 hr (or 12 hr) intervals
in urine? It would provide better comparison with rates of excretion of
2.4?0 and in humans where only small amounts, if any, were in
conjugated form.

Secondly, fecal samples should be analyzed for longer than the first two
days after dosing to see whether fecal excretion was delayed, at least for
those subjects where total recovery in urine was less than 752 (subjects

5 and 6). It is possible that silvex taken up by the second compartment

in the body would end up in feces. However, the first two fecal samples
may have been taken as early as 0.5 and 24.5 hr after dosing which doesn't
allow much time for uptake, possible conjugation in the liver or elsewhere,
and delayed elimination as conjugate(s) in feces.

Thirdly, Figure 2 for excretion in urine should be based on silvex plus
silvex conjugate to represent total rate of elimination of the chemical
from the body. Differentiation should be made between "metabolism OPERATING UNIT OF THE Dow CHENIICAL COMPANY .

M. W. Sauerhoff August 10, 1976

which the parent chemical can be regenerated unchanged from the conjugate,
and true metabolism in which the parent molecule is irreversibly altered
(such as by hydroxylation of the ring, cleavage of the ether link to
release the trichlorophenol, etc.). Similarly, hydrolysis of silvex
ester(s) to give silvex and silvex conjugate(s) is not comparable to true
metabolism/degradation of the herbicide molecule.

It will be interesting to see whether reanalysis of the data for silvex
plus silvex conjugate will take care of the biphasic pattern of elimination
developed by the computer. This pattern is not readily apparent from the
plot of silvex excretion by the eight subjects (Figure 2). Perhaps total I
excretion will fit a one compartment model as found for and 

Were any studies done on ease of dissociation of the conjugate(s)? As far
as I can see, the samples were simply adjusted to pH 1 or 13 and refluxed
to hydrolyze any and all conjugates. 0n the other hand, a slight change
in pH might have been adequate to release the silvex. If the conjugates
are very labile, this could account for the variability in preportion of
free vs conjugated silvex in the urine among the various subjects.

The apparent difference between silvex and and may also be
due to the fact that the silvex administered was a racemic mixture of
optically active isomers. Perhaps one isomer forms soluble salts like
2,4wD and whereas the other isomer may preferentially form a
conjugate with some optically active amino acid. Only one isomer of
silvex has herbicidal preperties according to Loos in a book issued
recently (Kearney et a1. 1976).

As we discussed, I would greatly appreciate having recalculated half?liveES)
based on total excretion of silvex in urine as ?293 as possible. I am
currently finalizing a paper on comparative metabolism of phenoXy herbicides
in animals for presentation at the ACS meeting in San Francisco on

September 2. One slide deals with the three studies conducted by Dow in
humans, comparing the excretion pattern and rates of elimination for 
and silvex following single oral doses of 5, 5, and 1 mg/kg,
respectively. It would help if the half-lives were all based on total
excretion.

Joe LeBeau has suggested that we meet soon to discuss the data generated

in this study. Milt Getzendaner is commenting separately On the need for

more actual analytical data, such as on analyses of individual samples.

We are also concerned about inclusion of a paragraph on page 16 which
postulates that "the fraction of silvex not recovered in some subjects may
represent metabolism of silvex and excretion in urine or bile of a heretofore
unidentified The report also suggests that studies without

a tag are of limited value. Such statements should not be made in

reports which may be submitted to government agencies or used in court actions.

 (we; :41 lav-52'

Marguerite L. Leng
Government Registration
Health and Environmental Research

abc

164i

DOW CHEMICAL USA

 

 

MIDLAND, MICHIGAN 48640

November 8, l978

Irving J. Selikoff, M.D.
Environmental Science Laboratory
Mt. Sinai School of Medicine .
lDOth Street and 5th Ave.

New York, N.Y. 

Dear Dr. Selikoff,

It was good to talk to you yesterday. This letter is to confirm with
you that Dow scientists are pleased to accept your invitation to come
to Mt. Sinai and present a seminar on phenoxy herbicides on Wednesday,
December 20, l978.

It is my (our) understanding that you wish us to present the Scientific
data we have on and related substances. The purpose

is to acquaint you and your staff with the data so that you will be .
better able to plan and execute the proposed epidemiologic study in the
Pacific Northwest. 

The people we have selected to present the seminar are:

John Davidson, Technical Advisor to the Director of Research and
Development, Agricultural Chemicals Department.

Bernard Schwetz, Director of Dow's Toxicology Research Laboratory
in Health and Environmental Research.

Richard Kociba, Head of Pathology in the Toxicology Research
Laboratory.

Benjamin Holder, Medical Director for Dow USA and Director

of Biomedical Research in Health and Environmental Research
Warren A. Crummett, Technical Manager of Analytical Labora?
tories.

V. K. Rowe, Director of Toxicological Affairs, Health and Environ?
mental Sciences (I may come along as moderator or whatever).

You asked if we had any objection to your inviting a few scientific
colleagues from such places as NIEHS and NYU. Certainly not.

AN UNIT OF THE DOW CHEMICAL COMPANY

Le'saao?oc

18419

.

 

I. J. Selikoff . 4? November 8, l978

I mentioned to you that Walter Melvin Jr. M.D., M.P.H. and of the
Institute of Rural Environmental Heath, Colorado State University at

Fort Collins might well be willing bucontribute. Dr. Melvin is well
informed and has had a-svealth of ham experience in the phenoxy herbicide
area. He served with the Air Forcef?r many years and has been directly
involved in Viet Nam, with the dispai? of the Air Forces? controversial
inventory of Agent Orange, with theiiyeso incident and presently is
engaged in an environmental or epidaiological study of some sort in

the Oregon-Washington area.

You asked what papers your-people cnl'd benefit from reading before the
seminar. I suggest the fellowing:

l. Toxicology of Phenoxy Her?iides and Hazard Assessment of their
Use in Reforestation by Funk N. Dost. 

2. Long-term Hazards of Polyc?orinated Dibenzodioxins and Poly-
chlorinated Dibenzofurans. WHO IARC Technical Report No.
78/00], a report of the Jd?t Working Group.

3. Chlorinated Phenoxy Acidsand Their Dioxins. Ecology Bulletin
No. 27 from.Royal SwedishAtademy of Sciences, C. Ramel 

4. Embryo Problems Posed by u: Seveso Accident. H. Tuchmann-
.-Duplessis. M.D., LeConcoussMedical No. 44, Nov. 26,.1977.

I have included copies of the Titleppge of the first three of these so
as to help in identification. Thereare, of course, hundreds of papers
but these reviews will give a good eerall perspective and references

to specifics. I am enclosing a copyuf a translation of number 4 because
I did not expect that-you had seen ttor might not be able to get it in
time and I do think it is important; 

I hope I have answered all the questins you asked. If you have specific
items you would like to have discusd. let me know and I will do our
best to include them.

With best personal regards,



V. K. Rowe, 

Director, Toxicological Affairs
Health and Environmental Science
l803 Building

(5l7) 636-2376



.bcc: E_Blair P. Gehring
En6105ure H.6rummett B. Holder
J.Davidson R. Koc1ba
- J.Donalds Schwetz

P.5. Asof now we are planning to take the Dow plane
I will

Lto?ew York and back on December 20.
:91: you more details later.

BESBZO
MOO

 

mt g?v?I 

 

 

 

2540

TOXICOLOGY OF PHENOXY HERBICIDES AND


Frank N. Dost, D.V.M.

Department of Agricultural Chemistry and
Environmental Health Sciences Center
Oregon State University
Corvallis, Oregon 97331

February 1978

 

a. 1100 M00

18421

TOXICOLOGY 0F PHENOXY HERBICIDES AND


Introduction . . . . .

Toxicology of TCDD . . . . . . . . . . . . . .

History of Human Exposure . . . . . . . . . . .
General Toxicity in Laboratory Animals . .

0

Pathologic Morphology Resulting From TCDD Intoxication

Effects of TCDD Upon Reproductive Functions . .
Carcinogenic and Mutagenic Potential of TCDD . . .
Absorption, Distribution, Metabolism, and Excretion
Behavior of TCDD in the Physical Environment and in

Submammalian Species . . . . . . . . . . .
Induction of Microsomal Enzymes . . . . . . .

2,415-General Toxicity in Laboratory Animals . . . . . .
Effects of on Reproduction . . . . . . . .
Carcinogenic and Mutagenic Potential of .
Absorption, DiStribution, Metabolism, and Excretion
Behavior of in the Environment and Effects on

Submammalian Species . . . . . . . . . . . . . .

.

Toxicity to Humans . . . . . . . . . . . . . . . .
General Toxicity to Laboratory Animals . . . . . .
Effect of on Reproductive Function . . . . .
CarcinOgenic and Mutagenic Potential of . . .



Absorption, Metabolism, Tissue Distribution, and Excr

0f 2, 4-0 . a 0
Behavior of in the Environment and EffeCts on
Submammalian Species . . . . . . . . . . . . . .

 Biological Effects of Silvex and Its Derivatives .
Effect of Silvex on Aquatic and Invertebrate Species
Metabolic Fate of Silvex in Mammals . . . . . . . .

References . . . . . . . . . . . . . . . . . . . . .

etion

of TCDD .

nr?w Mar?



16432

TOXICOLOGY OF PHENOXY HERBICIDES AND


Frank N. Dost, D.V.M.

Introduction

The phenoxy herbicides 2,4,S-trichlorophenoxy acetic acid 
2,4-dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxypro-
picnic acid (silvex) are important agents for reforestation site prepara-
tion and for release of conifer plantings. To be used, the compounds
must be inserted into a segment of the environment that may be occupied
by humans or may have a potential for migratiOn to points of human con-
tact or consumption. This document is a review of the pertinent litera-
ture describing biological effects and environmental behavior of the
three herbicides, and of an extremely
toxic contaminant found in small quantities in and silvex.

For various reasons, the greatest public attention has been focused
on and its unique contaminant, TCDD. Since 1969 public and sci-
entific attention to TCDD has overshadowed the specific concerns about
which has no TCDD, and about and silvex, which are con-
taminated.

The unusual nature of the TCDD contaminant has made a special case
of the herbicide and has made it the subject of an unusually
vehement public, political, and scientific controversy. An enormous
scientific effort has evolved in an attempt to learn about the unique
character of the compound, and to provide a rational basis for regula-
tory decisions. Hopefully, this effort to bring the existing literature
into focus will aid in the decision-making process.

As matters presently stand, there is a division in the collective
public and scientific minds on the safety and benefits of use of the
phenoxy herbicides. As in any controversy, both views include unreason--
able people, but the bulk of the argument is among sincere citizens and
competent scientists on either side. It is my hope that the reviews and
assessments in this document are dispassionate and without bias; other-
wise it will be of limited utility.

In preparing this review, emphasis has been placed on information
about the toxic properties of the re5peCtive agents. An assessment of
hazard necessarily considers along with biological effects, factors govern-
ing the probability of exposure, and the probability that the chemical
once contacted will enter the organism.

With the exception of the section on TCDD, I have chosen to touch
only upon the environmental behavior of the compounds. This
information about phenoxy herbicides is well established and is much
less complex than the behaviOr of TCDD. A more thorough treatment of
the environmental behavior of TCDD is necessary here because: 1) this
contaminant is clearly the most important of the four compounds under

18423

be

DU 

DOW

 

0'01"  

?study; 2) the high toxicity demands thorough understanding of the com-

pound as it exists outside the target of toxic effect; 3) the environ-
mental chemistry of TCDD is incompletely studied and subject to some
controversy, whereas that of the herbicides themselves is well estab-
lished.

The organization of this review and aSSessment includes sections re-
viewing the scientific literature on TCDD, 2,4-0, and 
(silvex), an assessment of the hazard potential of their distribution in
the environment, recommendations on some aspects of methods of use of
the herbicides and recommendations of needed research. A shortened ver-
sion of the review and assessment is also attached. Organization within
the review differs among the chemical sections because of differ-
ences in research emphasis or the character of the agent itself. In cer-
tain instances, for example in considering the enzyme induction capacity
of TCDD, an area may be dealt with that is not entirely germane to the
issue of hazard but of considerable importance in understanding the biology
of the compound.

A number of useful reviews of interaction of phenoxy herbicides or
TCDD with animal systems have been or are being published. The most
recent one by Leng (1977), discussing metabolic fate of phenoxy acids in
domestic animals, Mercier (1976) in response to the Seveso accident which
spread large amounts of TCDD through several Italian communities, and
McQueen et a1. (1977) refuting alleged association between herbicide use
and fetal abnormalities in humans. McKenzie et a1. (1975), Norris et a1.
(1972), Buhler (1977), and the EPA position paper arising from the Dioxin
Working Group (presently in draft) are also all of considerable value.

TOXICOLOGY OF TCDD

History of Human Exposure

 

Probably the first clinical description of human TCDD exposure arose
from the study by Kimmig and Schulz (1957) of workers in chloroPhenol
factbries in Germany. The principal was a persistent skin lesion
(chloracne; so?called because it is characteristic of a number of related
componnds and was at one time thought to be caused by free chlorine.

The aetive agent was established as TCDD, and animal studies showed
that the same lesions appeared on rabbit ears after application of as
little as 0.002% TCDD (Kimmig and Schulz, 1957). High topical doses and
oral doses of 50 ug/kg or greater caused liver necrosis. A similar in-
dustrial intoxication in France was reported by Dugois and Colomb (1957).
Pathology of the skin lesions is well described by Kimbrough (1974).

A study of 29 phenoxy herbicide workers by Bleiberg et al. (1964)
disclosed chloracne and a high frequency of and tissue
deposition of related with hyperpigmentation and skin fragility.
This assemblage of is commonly called cutanea tarda.

The excessive production and excretion of apparently results
from a hyperactivity of the enzyme 6-amino levulinic acid (ALA) 
the first and rate limiting step in formation. are

inset

eventual components of heme proteins, such as hemoglobin and cytochromes.
Effects on ALA and other enzyme systems will be discussed in
another section. Poland et al. (1971) examined the same factories later,
after somewhat more satisfactory industrial hygiene measures were estab-
lished. Among the 73 male workers observed, no clinical was
apparent even though some degree of chloracne still persisted in most of
the subjects. There was also apparently a significant effect
as measured by the MinneSOta Multiphasic Personality Inventory.

 

The absence of and persistence of chloracne in the later
study has led Poland et a1. (1971) to conclude that while both may be
caused by TCDD, the mechanisms are probably dissimilar.

Several severe accidental exposures to TCDD have occurred. In several
cases, runaway chlorophenol plant reactions caused widespread exposures
within the factory or in the surrounding community. An accident in
Germany in 1953 exposed a large number of workers, said to have suffered
liver, kidney, splenic, eye, cardiovascular, and nervous A
worker who apparently was in extensive contact with a contaminated seg-
ment of the plant five years after the accident became ill and later died.
An explosion in a Dutch plant in 1963 released 30?200 of TCDD into the
main gallery of the factory exposing 50 people. In the subsequent two
years, four died, but no clear association with TCDD intoxication could
be shown (Hay, 1976). An explosion in a British plant in 1968 resulted
in 79 cases of chloracne (May, 1973). In Czechoslovakia a pentachloro-
phenate production stream overheated under pressure, producing large amounts
of TCDD (Jirazek et al., 1974). (It should be noted in this and some ear-
lier German work a different numbering convention was used. 
and are the same compound.) Of 80 exposed workers, 76 devel-
oped chloracne, cutanea tarda, disorders of plasma lipids,
hepatic damage, neural lesions, and neurasthenia.

An explosive reaction in a plant at Seveso, Italy, in 1976 scattered
TCDD, apparently in kilogram amounts, over a wide area that included sev-
eral communities. Details are still not widely disseminated but great
numbers of animals were killed and many peeple were made ill. A review
of the incident was prepared by Mercier (1976) a few months after the
incident but the slow onset and persistent effects did not permit a de?
tailed assessment of the damage. Since that time the Seveso disaster
has been given unprecented publicity and as might be expected this tragedy
has been sensationalized in press coverage, in part because there may
have been an effort to minimize or cover up the extent of the problem.
There were apparently defects in the administration of the plant that
may have resulted in the accident itself, and management of public warn?
ing and information early after the accident was questionable. Further
contributing to the extensive public attention and sensationalizing of
the incident is a virtual absence of objeCtive reporting in the scien-
tific literature. As far as I am aware, there has been no scientific
evaluation of the aftermath in publication since the review by Mercier
(1976), shortly after the accident.

In the U.S., TCDD extracted from hexachlorophene production was
stored in oil, then mistaken for waste lubricating oil and sprayed on
several horse arenas and farms in Missouri, for dust control. The

 

incidents occurred in Spring and summer of 1971. On one horse breeding
farm, 62 of 85 horses exercized in the sprayed area became ill and 48 died,
the last in January, 1974, two and a half years later. HUndreds of birds
and rodents died, along with a number of dogs and cats. Several children
were exposed through play in the arena soil. One developed a severe hemor-
rhagic cystitis and others showed clear evidence of chloracne. All re-
covered. The TCDD concentration in the soil was later established at more
than 30 ppm, a truly massive amount. (This concentration amounts to 120 lb
per acre per foot of depth, assuming 4 106 lb The de-
tails of the latter event are described in detail by Carter et a1. (1975)
and Kimbrough et a1. (1977).

All of these exposures were to a mixture of TCDD with other agents.
Possibly the only published account of exposure to pure TCDD is the des-
cription by Oliver (1975) of laboratory contact by three individuals as-
sociated with experiments. The individuals all had utilized
routine protective procedures but sufficient contact developed nonetheless.
The exposures were not sufficient to cause or liver damage
but serum cholesterol was raised. In two cases personality changes and
neural disorders developed two years after exposure. The amount of TCDD
contacted is not known.

General Toxicity in Laboratory Animals

 

This secrion is a review of data on effects of TCDD other than repro-
duction and teratology, carcinogenesis, and its enzyme inducing properties
upon mammals. These will all be considered independently, as will studies
of tissue diStribution and metabolism.

Schwetz et a1. (1973) have determined lethalities for several species.
The guinea pig is exceedingly sensitive, with an LDso of only 0.6 ug/kg;
rats are less sensitive; L050 for males is about 20 ug/kg and for females
the L050 is in excess of 40 ug/kg. Rabbits are still less sensitive at
more than 100 ug/kg. Dogs were found to require even higher effective
doses. In these studies, responses of mice were highly erratic and were
not included. McConnell et al. (1977a) have determined a single 30 day
LDSO for mice of 284 ug/kg, and 2 ug/kg for guinea pigs.

An important characteristic of TCDD is a very long delay before
lethality, often 2?3 weeks and occasionally-7-8 weeks.

The pathology associated with TCDD toxicity may include thymic atrophy,
liver necrosis, and degeneration in the kidney and thyroid. The guinea
pig is unique in that it suffers limited hepatic damage, even at doses
that cause substantial thymus depletion (Gupta et al., 1973). The degree
of thymic atrophy is generally dose dependent; the studies of Gupta et al..
(1973) did not establish a zero effect dose, but the lowest dose in the
rat of 1 ug/kg/day for 31 days caused moderate thymus and liver damage,
and kidney and thyroid degeneration. Guinea pigs treated with 0.2 ug/kg
weekly for eight weeks had limited thymus damage and no other pathology.
Studies of guinea pigs at_lower doses suggest a no observed effect 1eVel
at 0.008 ug/kg/weekly for eight weeks, a total of 0.064 ug/kg, although
there was a slight decrease in at that dose (Vos et al.,

1.15425

1973). Animals given tetanus toxoid at the fourth week to measure humoral
immunity were found to have somewhat lower thymus weights than controls,
although the difference was not significant.

According to the data of Kociba et a1. (1976) the no observed effect
level in rats given TCDD in the diet 5 days a week for 13 weeks is 0.001
ug/kg/ day, totaling 0.065 ug/kg, almost identical to the value found in
guinea pigs. Again, thymus weight appeared to be the most sensitive param-
eter. The Kociba study dealt with a very broad range of clinical, chemical,
and hematoIOgical measurements. At a dose of 1.0 ug/kg/day, 2 of 10 rats
died and weight gain decreased sharply, numbers and volume
decreased, reticulocytes increased in males. In females thrombocytes
decreased and total leukocytes increased. Differential counts were not
altered. Urinary creatine and 6 amino levulinic acid (ALA)
all increased. Serum bilirubin, BUN and alkaline phosphatase also were
elevated. At an intake of 0.1 ug/kg/day red cell numbers and volume were
still decreased in males and ALA, bilirubin, and al?
kaline phosphatase was elevated in females. No changes in blood parameters
occurred below these levels.

 

1100

In a similar study Zinkl et al. (1973) found no changes in SGPT,
bilirubin, or after 31 days of treatment at 1.0 ug/kg/day and
no change in SCOT, cholesterol, or serum protein at 0.1 ug/kg/day. There
was a modest depression in blood glucose at the lower dose rate.

Mice treated with 1.0 ug/kg/week for six weeks were found to have
depleted thymus weight (Vos et al., 1974) but no changes were evident at
the lowest dose, 0.2 ug/week. Hematological changes occurred only at a
dose of 25 ug/kg/week, for six weeks. Some changes in serum proteins did
occur at an intake of 0.2 ug/kg/weekly. Unfortunately, there is no data
suggesting the magnitude of the no-effect single dose in any species.

Studies in primates have been limited. McNulty administered TCDD at
a rate of 20 parts per billion in the diet to a single monkey and caused
its death in a few days. A 10-fold lower concentration permitted survival
for about two months. Total intake was estimated as about 6 ug/kg (McNulty,
1977). A study of eight female Rhesus monkeys fed 500 parts TCDD per tril-
lion has recently been completed in the laboratory of J.R. Allen. The
animals showed dermatitis and menstrual irregularities within six months
after initiating treatment. One monkey died prior to the seventh month and
by nine months of exposure, all survivors experienced a severe anemia and
leuk0penia. Food intake was normal but all of the animals lost weight.
After 11 months, five of the eight monkeys died, all with severe anemia and
destruction of blood forming tissues. The total dose was estimated at
about three ug/kg (Allen et al., 1977a, 1977b) or about .3 ug/month.

A number of questions arise from these data. The monkey studies are
limited, but they do suggeSt the possibility that the lethal dose is
similar regardless of the time over which it is administered. The im-
plications are quite interesting. If it is true that TCDD dosage is time
independent in the monkey, then the effect is truly cumulative. No chemi-
cal is known to have such properties, and if TCDD behaves in this fashion,

18427

 

any estimate of acceptable dose must assume additive effects over long
periods. TCDD does not remain in the body of monkeys (Van Miller et al.,
1976) or other species (Van Miller et al., 1976; Fries and Marrow, 1975;
Rose et al,, 1976). A cumulative effect, if it exists, must then entail
irreversible effects remaining after the molecule leaves active sites.
TCDD is apparently not metabolized to a significant extent (Rose et al.,
1976; Vin0pal and Casida, 1973; Van Miller et al., 1976) so it is conceiv?
able as well that a single molecule could interact several times before
leaving the body. A possible mechanism may be intercalation of the planar
TCDD molecule into DNA (Hussain, 1973), production of misinformation and
departure of the agent to interact at another DNA site. 

Data in other Species, e5pecially the guinea pig do not support the
idea of extensiVe cumulative effect. A dose rate of 0.2 ug/kg/weekly
for eight weeks provided a total dose almost three times the LDso,
but no deaths occurred and organ changes, while measurable, were not
severe (Vos et al., 1973). Rats also accepted about three times the
L050 of 20 ug/kg over 13 weeks, without lethality. Some mice, on the
other hand, died at a total dose less than the acute LDso, distributed
over a four week period (Vos et al., 1974).

There are a few other biological responses to TCDD that appear after
high doses of from 10-25 ug/kg. One of the more interesting is a de?
pressed capacity for biliary excretion. TCDD sharply decreases removal
of PCBs from the liver via the bile (Yang et al., 1977) and decreases
plasma clearance of ouabain into bile (Hamada and Peterson, 1977). In
the latter work, steroidal inducers of microsomal enzymes reversed the
TCDD effect. Whether this effect has porential significance in the
response to secondary toxicity can only be speculated upon. In any case
the doses required for effect are quite high and probably not attainable
under realistic environmental conditions.

Because TCDD causes increased microsomal foreign compound metabolizing
activity in the kidney cortex (Fowler et al., 1975, 1977), Pegg et a1.
(1976) evaluated proximal tubular function in intoxicated rats. They
found little change at any but large doses (25 and 50 ug/kg).

There have been few observations of metabolic changes caused by TCDD.
Incorporation of 3H-sodium acetate into liver lipid fractions seems to be
impaired at TCDD doses above 0.1 ug/kg (Cunningham and Williams, 1972),
but the significance of this effect has apparently not been explored fur-
ther.

Evidence of thymic atrophy after TCDD treatment has stimulated more
direct study of effects upon the immune responses. Vos and Moore (1974)
and Vos et a1. (1973) found depletion in the cortex of the
thymus and depressed cellular immunity in rats and mice exposed during
gestation and the post-natal period. Guinea pigs were similarly affected
(Vos et al., 1973). Thigpen et a1. (1975) attempted to translate the
effect into terms of infectivity and found that doses of ug/kg or more
increased mortality and time to death of mice infected with Salmonella
bern. Course and severity of disease due to pseudorabies virus was not
altered by TCDD. With all the Study of TCDD effects, no pattern of

16428

MOO

pathology, tissue distribution, and biochemical or physiological change

 

 

Gag .
has emerged that is consistent enough to suggest a mechanism of lethal CDQ -
effect. McConnell et al. (1977a) quote papers in preparation by Van 
Logten et al. which show no association of death in TCDD intoxication 
With adrenal or pituitary hormones or malnutrition. 
Pathologic Morphology Resulting From TCDD Intoxication 

Much of the more pronounced tissue change resulting from TCDD has
been observed following high acute doses of the compound. There is
some question whether the acute damage seen represents that observed in
a low level longer term experiment or in a field exposure.

Before TCDD as such became an issue Norback and Allen (1969) studied
the membrane changes in liver following ingestion of hexachlorohexahydro-
phenanthrene, a constituent of so-called "toxic fat." The lesions found
under electron and light microscoPy anticipated the damage later to be
found after TCDD. Most striking was a great increase in the smooth endo-
plasmic reticulum (SER), with reorganization into a concentric arrange?
ment, and the rough ER became folded and rolled over a period of days to
form dense arrays of concentric separate, not spiraling, paired membranes.
The authors speculate that SER proliferation in response to intoxication
is related to acceptance of enzymes by the RER. The in?
creased membrane mass may also be a means of sequestering chlorinated hy-
drocarbon in the lipid phase of the membrane in close proximity to foreign
compound metabolizing enzymes. (This paper contains very useful schematic
representations of the membrane development sequence.) The same authors
later reported similar leSions in rats following administration of 1 ug
TCDD/kg/day for 21 days (Norback and Allen, 1973). They found hypoplasia
of tissue and bone marrow with progressive anemia and leukopenia.
The resultant reduced resistance caused substantial losses due to infec-
tion. Monkeys and chickens in the same study were much more sensitive
in the latter parameters than rats. They also developed extensive fluid
accumulation in all body cavities, probably due to decreased osmotic
activity of blood due in turn to serum protein decreases. (Relative
sensitivity of chickens has also been nOted by Greig et a1. (1973).)
Monkeys were subject to extensive skin lesions, apparently corresponding
to the chloracne of humans, but rats and chickens did not Suffer skin
damage. Monkeys and chickens also exhibited degeneration of the testes

and decreased activity of seminiferous tubules (see also Allen et al.,
1975).

Large doses of TCDD (single doses of 50 or 100 ug/kg; 16 to 31 daily
doses of 10 ug/kg) to the rat caused severe liver and thymus damage, with
icterus and disseminated hemorrhages, especially in the myocardium. The
liver cells increase in size and cellular regenerative attempts occur.
Renal tubule cells became vacuolated. No pathological change occurred at
doses of 0.1 ug/kg/day over 31 days, at 1.0 ug/kg weekly for 6 weeks or 5
ug/kg in a single dose (Gupta et al., 1973). Fowler et al. (1973) studied
the progressive changes in rats over 28 days following a single dose of 
or 25 ug Increased SER, roughly dose responsive, was evident by
day three, especially in cells adjacent to biliary drainage. Both RER and
SER were greatly increased by day 9, SER was almost at normal levels by day

immas

.216, and both were indistinguishable from those of control animals by day
Jones and Butler (1974) carried out a similar experiment with rats
V-given a single 200 ug/kg treatment. (A massive dose, considering the mini-
mum effective dose.) Most change was found in centrilobular cells, with
degeneration and necrosis of cells evident within a week

after treatment. They suggest that the multinucleate cells seen in in?
creasing numbers through two months after treatment are formed through
membrane disturbance and coalescence of hepatocytes. By 10 weeks some
recovery was evident but fibrosis of central veins and sinus dilation

still prevailed. No mention of lethality was made, although the dose

was on the order of five (female) to ten (male) times the LDSO. Longer
term treatment of rats (0.001-1.0 ug/kg/day, 5 days weekly 13) caused
almost complete thymus involution at a dose rate of 1.0 ug/kg/day, somewhat
decreased cortical thymocyte and cell numbers after 0.1 ug/kg/
day. Splenic morphology was essentially unchanged at 1.0 ug/kg/day.

The latter dose also caused some gross edema, and one of five males had
decreased spermatogenic activity. Females at the highest dose tended
toward formation of cuboidal epithelium in the uterus and decreased size
and numbers of corpora lutea. Ovarian tissue was alSo abnormal. At all
lower doses no effects were observed (Kociba et al., 1976). Two year 3
continuous feeding studies at TCDD intakes of 0.001 to 0.1 ug/kg/day have-
just been terminated and histopathologic evaluation is not complete.

Gross examination showed that in some rats given 0.1 or 0.01 mg 
day there were increased numbers of nodules in the liver (R.J. Kociba,

Dow Chemical Company, preliminary status report; personal communication).

 

Lesions generally similar to those seen in rats were found in mice
given single oral doses up to 200 ug/kg and weekly doses up to 25 ug (Vos
et al., 1974). At a dose rate .1 ug/kg/week for 6 weeks decreased thymus
weight occurred. At 25 ug/kg/week, serum globulins were decreased, and
and bile duct epithelial proliferation occurred. Similar changes
were also reported following treatment of mice with an LDso (30 day) TCDD
(McConnell et al., 1977a).

McConnell et al. (1977b) treated rhesus monkeys with heavy single
doses of TCDD (in excess of 70 ug/kg) which were fatal in eVery case.
Integumental changes were pronounced, anemia and and an in-
creased neutrophil count followed treatment. Liver, adrenal and kidney
appeared to increase in weight relative to body weight, but this effect may
have been due to general wasting. With the skin hyperplasia, similar
changes occurred in the epithelium of some hollow organs. The major dif-
ference from effects in similarly treated rats was the absence of sig-
nificant liver lesions.

Chronic low level intoxication (500 in diets, about 0.01 ug/
kg/day) of monkeys until death at 7-12 months caused anemia and pancyto-
penia, with extensive hemorrhage. Bone marrow and tissues
were hyp0p1astic, and epithelial structures were hyperplastic (Allen et
al., 1977).

The pathological changes resulting from absorption of TCDD by

horses and cats during the Missouri horse arena episode have been recently
described in detail by Kimbrough et a1. (1977). The moSt pronounced and

154307

I
a

 
consistent findings were the generalized hepatic fibrosis and necrosis ?g
and gastric ulceration. There was an appreciable incidence of tubular


degeneration and abscesses in the kidney and capsular thickening of the 3
spleen. Lesions in cats were generally similar. a;


LD
U1

Effects of TCDD Upon Reproductive Functions-

Of the many toxic responses to TCDD, the potential for teratogenicity
has received perhaps the greatest notoriety. The early findings of 
teratogenicity raised a considerable public consciousness and the finding
that the herbicide was contaminated by high leVelS of TCDD, also found to
cause fetal malformation, focused additional attention on the contaminant.
In evaluating this aspect of TCDD hazard it should be noted that the
teratogenic potential of a toxicant can be considered in two ways. The
absolute teratOgenic dose of TCDD to the pregnant animal on a body weight
basis is very low, as is true of all TCDD effects. A more realistic way
of relating the teratogenic potential, however, is by comparison with the
dose required to cause general toxic effects in the mother. For example,
in the guinea pig, TCDD teratogenesis has not been studied because the
teratogenic dose is apparently higher than the lethal dose.

This section is limited to studies of relatively pure TCDD upon
reproductive quCtion in experimental animals. There is reference else-
where to reproductive studies of and silvex, which contain at least
some TCDD.

Teratogenic defect resulting from TCDD are rather specific in experi-
mental animals; the principal effects are increased frequency of cleft
palate (reported only in mice) and an abnormality in which the central
collection region of the kidney becomes enlarged, with an associated fluid
accumulation (Neubert et al., 1973). Limited limb deformation may be seen
in a few animals (Sparschu et al., 1971). Intestinal hemorrhage is also
seen; this effect is not teratogenic but rather is direct toxic consequence
of fetal exposure to TCDD. Fatty degeneration of fetal livers is in the
same category (Becker, 1974) as is subcutaneous edema and delayed ossi-
fication.

In teratogenicity studies of rodents the usual procedure is to ad-
minister intoxicant at some point during, or throughout days 6-15 of preg-
nancy, which covers the organ forming period. Sparschu et a1. (1971)
found that the lowest effective dose of 1.25 ug/kg/day caused some fetal
mortality, resorptions, and intestinal bleeding. Skeletal abnormalities
were limited to delayed ossification. Renal defects tended to occur at
about the same frequency regardless of dose. These changes are all a
result of direct toxicity to the fetus and are not developmental. In-
testinal hemorrhage has been reported in rat fetuses at maternal dose
rates of 0.25 ug/kg/day (Khera and Ruddick, 1973). At higher doses up
to 3 ug/kg/day the same kind of effects occurred with greater frequency;
0.5 ug/kg/day caused decreased maternal weight gain and higher doses
resulted in severe toxicity.

Courtney and Moore (1971) examined rats and three mouse strains and
found that a dose of more than 1 ug/kg/day was required to produce kidney

18431

 

 

-gestation.

defects in rats, and the cleft palate and kidney changes in mice appeared
at about 3 ug/kg/day, over days 6-15 of gestation. The C5731 mouse strain
was muoh more sensitive than the others tested. A somewhat larger group
of NMRI mice showed a cleft palate frequency of about 3% (Neubert and
Dillman, 1972), similar to the low frequency groups of Courtney and Moore.
The period of maximum sensitivity for TCDD is at the eleventh day of
gestation, as distinguished from dexamethasone, for example, which is

most effective at day 13 (Neubert et a1., 1973).

Continuous administration of 0.001 ug/kg/day to male and female rats
for 90 days prior to mating and through weaning of produced no
effect on fertility (MUrray et al., 1977). The study continued through
three generations, with each preceeding generation sacrificed at weaning
of its the third generation was maintained until two years
from the beginning of treatment of the fa generation. Toxicity and poor
litter survival dictated termination of feeding at 0.1 ug/kg/day; 0.01
ug/kg/day caused decreased fertility in generation litters were smaller and growth and survival were decreased
after treatment at 0.01 ug/kg/day, but no pathological change was observed
in liver, kidney, or thymus.

More recently, Smith et al. (1976) have treated mice at doses
ranging from 0.001 ug to 3.0 ug/kg/day through days 6~lS of gestation.
There was no maternal toxicity at any dose. Cleft palate appeared at
1 ug, renal dilatation appeared at 3 ug/kg/day. No significant effects
occurred at 0.1 ug. -

A recent report by Courtney (1976) described results of treating
CD-1 mice with high doses of TCDD (25-400 ug/kg/day) on days 7-16 of
These very high doses caused no maternal deaths during the
gestation period, but resulted in high fetal mortality. The nature of
observed terata were typical of TCDD intoxication, but the enormous
doses render the study useless for hazard evaluation. However, the
substantially greater incidence of terata after subcutaneous administra-
tion, compared with that following oral TCDD suggests that hepatic metabo-
lism may affect TCDD more than has been believed. A dose rate of 25 ug/
kg/day orally caused 3% incidence cleft palate; the same dose subcutane-
ously caused 82% incidence of cleft palate.

There has been little research on effects of TCDD on primate repro-
duction. Experimental animals are in limited supply and the numbers of
are so low that a satisfactory study is almost impossible.

Allen et al. (1977) fed TCDD to eight female rhesus monkeys at a con-
centration of 500 in the diet, which provided an intake of about 0.3
ug/kg/month. Skin lesions, endocrine, and menstrual changes occurred in
all of the subjects by 3 months. Six females of the eight were bred after 
7 months of treatment; one monkey died before mating and another was ex-
cluded from the reproductive study. Three of the animals conceived; two
aborted, and one completed a normal pregnancy. Two of the three barren
animals were bred four times, the other was bred twice. This dose rate is
on the order of 0.01 ug/kg/day and comparison with the rat study of Murray
et al. (1977) suggests that reproductive toxicity in monkeys may be some-
what greater than in rats, but is by no means prOportional to the lethal
dose. 

18432

10

Carcinogenic and Mutagenic Potential of TCDD

Van Miller and Allen (1977) have evaluated pathological changes in
rats fed TCDD in dietary concentrations ranging from 1 to 1000 ppb,
beginning at animal weights of about 60 g. EStimated weekly intakes of
TCDD ranged from 0.0003 ug/kg at a dietary level of to 2 ug/kg at
5 ppb. Actual intake of TCDD at dietary concentrations of 50-1000 
was not clear. At 65 weeks the abdominal viscera of surviving animals
were examined surgically and any observed tumors were biopsied. The diet
with TCDD was then continued through the 78th week and all surviving
animals sacrificed at 95 weeks.

 

There were 10 animals at each dose range; all those at 50-1000 
TCDD died by the fourth week. In the group given 1 there were no
tumors, but of those fed 5, 50, 500, 1000, and 5000 23 of 50 had
developed tumors of several types. No control animals for this or a
parallel experiment developed neoplasms. The great variety of tumors
suggested to the authors the possibility that TCDD is a promoter rather
than inducer of neoplastic activity, in view of the usual narrow spec-
trum of tumors caused by many chemical carcinogens. General pathologic
changes have been described in a previous section.

Two studies of carcinogenesis in rats have utilized continuous
feeding of TCDD over a two-year period. One program has been completed
but the analysis of histoPathOIOgy and gross tumor incidence have not
been completed. The other study, by Dow Chemical Company, has been
completed and a preliminary report filed with EPA. Dietary levels were
21, 210, and 2200 TCDD. The maximum dose caused increased tumor
incidence in some organs and decreased incidence in others. There was
Substantial mortality resulting from general toxic effects. The inter-
mediate dose caused moderate systemic toxicity but no tumors and the
lowest dose was without effect other than slight liver changes such as
induction of increased drug metabolizing capability. These latter
changes were seen only in females (J. Davidson, personal communication).
The reasons for differences between the Dow and Van Miller studies is
not apparent; it is paradoxical that the Dow work had a higher incidence
of tumors in control animals, while showing a lower incidence of effect
in treated animals.

Cytogenetic studies of TCDD by Green and Moreland (1975) suggested
that the dioxin does not have potential for producing chromosomal ab-
normalities in bone marrow. The dose schedule provided up to 15 ug/kg/
daily for days, with sacrifice on day S. A single injection experi-
ment with sacrifice at four weeks was also negative. Khera and Ruddick
(1973) were also unable to find mutagenic activity of 

Since it is not possible to obtain an absolutely TCDD-free prepara?
tion of the studies of the herbicide may also be of value in
considering TCDD effects. Effects of and TCDD on dividing Afri-
can blood lily endosperm cells are not explicitly pertinent to animal
studies, but the relation shown between herbicide and contaminant is of
interest. Highly purified at 10'4 was not effective, but 0.2

. 161MB

11

 

DOW

2

pg with or without and commercial all caused
dramatic mitotic inhibition and chromosomal aberations (Jackson, 1972)
(0.2 ug/L 200 ppt).

Hussain (1972) found that TCDD was somewhat/mutagenic using the
Ames procedure at doses that were intrinsically very toxic to the or-
ganisms. The authors suggested that the limited mutagenic activity
was by intercalation of TCDD into DNA. Seiler (1973) has also obtained
data suggesting that TCDD is mutagenic in the S. typhimurium TA 1532
strain. There was no indication of the concentrations necessary to
produce effects.

Absorption, Distribution, Lmtabolism, and Excretion of TCDD

The biological action of a foreign compound is highly dependent on
its logistics. A chemical must obviously be absorbed from the environment
in order to interact, and it must be carried in blood to cells and move
to or across cell-membranes to sites of reaction. The agent in its origi-
nal form may interact with sensitive systems, or it may be converted to
an.active derivative after entering an organism. Such conversion usually
takes place in the liver. A given organ or cell type may have a particular
affinity or sensitivity for a toxicant and thereby be singled out for at-
tack. The chemical nature of the agent or its product will also dictate
the extent and rate of excretion in urine or bile or respiratory gases.
Even with an active excretion mechanism such factors as sensitivity of
bladder epithelium or an active enterohepatic circulation may result in
toxic effects after elimination seems to have been accomplished.

In spite of the enormous capacity for induction of mixed function
oxidases (discussed below) TCDD is metabolized to a very limited extent.
Rats given a single dose of SD by stomach tube excreted
about 30% in feces during the two days following treatment, and then 1-2%
daily over the 19 days following, to a total of 53%. Urinary-14C was
about 13% and reSpiratory excretion totalled After the initial 2
days, total clearance half-time was about 17 days. Residual TCDD at
3, 7, and 21 days was greatest in liver and fat (Piper et al., 1973). The
long residence time and limited respiratory and urinary excretion suggested
that TCDD was essentially unchanged but this was not verified. As a gen-
eral rule lipophilic compounds are either sequestered in fat, eliminated as
conjugates in bile, or converted to a water soluble form and excreted by
kidneys. Absence of urinary excretion therefore implies limited conver-
sion. In a later study of rats given a lower single dose of ug/kg, TCDD
was identified in feces but not-urine, and again, liver and fat contained
the highest concentrations. was 31 days (Rose et al., 1976). In the
same investigation, repeated doses 5 days a week for 7 weeks resulted in
some urinary loss, but most TCDD was excreted into feces. In this program
of essentially continuous intake, was calculated as 24 days. The
residual radioactivity in the liver was identified as unchanged TCDD.

The report of Vinopal and Casida (1973) is suggestive that TCDD does not
metabolize only because after giving a dose of 130 ug to mice,

no mention was made of tritium in the urine. Allen et a1. (1975) found

that over a 25 day period about 4.5% of a dose of 50 ug appeared in

urine. The daily fraction of total intake excreted gradually increased,

which seems indicative of an unusually stable molecule, because at least a

modest exchange of tritium with body water should be expected. 

12

i
1.

   

The variety of indirect evidence indicating that TCDD is not metaboli-
cally altered must be considered in the perspective of other indirect evi-
dence that metabolic change may occur. Beatty and Neal (1975) have shown
that pretreatment with phenobarbital decreases and castration increases
TCDD toxicity. The former treatment increases and the latter diminishes
drug metabolizing capacity. The teratology study of Courtney (1976) also
is suggestive that liver metabolism of TCDD occurs. An oral dose of 25
ug/kg/day through the organogenesis period caused only 3% cleft palate
among the experimental fetuses, but subcutaneous administration of that
dose resulted in 82% cleft palates. In view of the apparent affinity of
TCDD for the increased endoplasmic reticulum following induction (Van
Miller et al., 1976; Poland et al., 1976), it is possible that induction
merely increases the capacity for sequestration of TCDD in a temporarily
non-active system, but the remarkable difference seen by Courtney is
difficult to ascribe to distribution differences only.

un?l-'
-

664. MOO 

The selective distribution of TCDD into liver and fat has also been
shown by Fries and Marrow (1975), Allen et a1. (1975), and Van Miller et
a1. (1976) in rats and by Van Miller et a1. (1976) in primates. -The
liver TCDD appeared to be sequestered on smooth endOplasmic reticulum
(SER), principally that induced by presence of the toxicant (Allen et al.,
1975). While liver storage of TCDD in monkeys was much lower than in rats,
the difference appeared to be due to the characteristic lesser prolifera-
tion of SER in the monkey that has been described by Van Miller et al.
(1976). Considering the difference between species in total induced ER
membrane, TCDD appeared to bind to endoplasmic reticulum to about the same
extent in both species (Van L?ller et al., 1976).

Unlike the behavior of other chlorinated hydrocarbons in fasted or
underfed animals, TCDD is not lost from adipose tissue with fat mobiliza-
tiOn (Allen et al., 1975). In the case of PCB, for example, the fat
soluble chlorinated hydrocarbon is liberated with mobilized fat and even-
tually may reaccumulate in the liver or other target organs. The reason
for the difference is not clear, but it seems possible that TCDD may non-
specifically partition into fat, then selectively bind to adipose cell
membrane protein. Liver TCDD residues do decrease during fasting, pos-
sibly because increased gluconeogenesis makes demands on available labile
protein including the endoplasmic reticulum to which the TCDD may be bound.
Hepatic subcellular distribution has been fUrther defined by Poland and his
associates (Poland and Glover, 1974, 1975; Poland et al., 1976; Nebert et
al., 1975); and by Chhabra et a1. (1974) in studies in which the geneti-
cally determined susceptibility for TCDD induced hydroxylase
induction was fDUnd to correlate directly with the extent of TCDD binding
in liver.

The for TCDD residence in the body has ranged from about 16-20
days for single dose experiments (Piper et al., 1973; Allen et al., 1975),
although Rose et a1. (1976) arrived at a figure of 31 days. When TCDD was
administered in the diet over 12 days at dose rates of about 0.5 and 1.5
ug/kg/day, was found to be 12 days for males and 15 for females (Fries
and Marrow, 1975). Rose et a1. (1976) calculated a half-time of 24 days
after a seven week feeding at dose rates of 1-0.01 ug/kg/day. The concentration of TCDD reaches a maximum at a given dose rate.
Fries and Marrow (1975) found that after 6 weeks of administration reten-
tion approached a steady state which would be 10.5 times the daily intake.
Rose et al. (1976) found similarly in 7 weeks that a steady state con-
centration of a little more than 10 times the daily intake could be pre-
dicted, agreeing remarkably well with the Fries and Marrow estimate.

The difference in distribution between primates and rats is of in-
terest. Seven days after equivalent doses of TCDD-3H, livers of adult
monkeys contained 0.09% of total dose/gm tissue, rats contained 4.54%
[Van Miller et al., 1976). In adipose tissue the relationship was monkey,
0.16, rat 3.46, suggesting that storage or retention is not strictly re?
lated to binding on inducible membranes. In terms of tOtal body distri-
bution, the rat liver contained 40% of the dose and that of the monkey,
10%. This is a much closer ratio than that of liver concentrations
Skin as a whole stored a much higher percentage of adminis-
tered TCDD, because of the much higher amount of adipose tissue, in spite
of a concentration ratio (3.5/0.16) that was also quite unfavorable.

 

Whether these differences bear any relation to differences in sensi-
tivity to toxic effects is not known. The difference in liver is at least
associated in amount and time with the much greater capacity for induction
in the rat. There is no data available to indicate whether the thymus of
species like the guinea pig, which is subject to more extensive TCDD in-
duced thymus damage, has a selective affinity for the toxicant as does the
liver in the rat. The thymus of infant monkeys is expected to be highly
active, but appeared less retentive compared with other tissues, while the 
adult rat thymus accumulated more TCDD/unit weight than most organs (Van 
l?ller et al., 1976), further confusing this issue because liver damage is
thought to be the primary lesion in the rat. 

Each of the single administration experiments utilized heavy doses of
TCDD, taking advantage of the lOng delay in onset of and death to
obtain improvement in radiochemical detection capability. Studies in
which 400 ug/kg [about 20 L050) were administered (Van Miller et al.,
.1976) produced liver concentrations of about 3.6 ppm. Kociba et al. (1976)
caused discernable but minor hepatic lesions at a total dose of 0.65 ug/kg
given over seven weeks. The expected concentration of TCDD in liver at
this intake is on the order of 20 (Rose et al., 1976). A ten-fold
lower dose caused no histopathologic damage. It is not surprising there?
fore that the beach mice studied by Young et al. (1974) which accumulated
300-500 TCDD in their livers, did not show liver morphologic lesions.

Behavior of TCDD in the Physical Environment and in Submammalian Species

 

The extremely high toxicity of TCDD and the very small amounts of the
agent present in and in the environment present a problem that is
different less in philosophy than in scale. There is a tendency on the
one hand to dismiss TCDD as a pollutant because the amount available at
any one point or even sector is minute, and on the other to assume that
any amount in excess of true zero residue is catastrophic,.because of
the enormous toxicity of the material. The amount of TCDD disrributed on
vegetation after a typical treatment is so small as to defy direct analysis
and the quest for residues in biological materials has forced analytical

14 15435

sensitivity closer to zero than has been the case with any other pollutant.
In spite of these unique characteristics, TCDD must be considered in

the same way as any other chemical: How much is present in the human
environment? How persistent is it? Can it move to unintended targets? If
so, will the amount assimilated by humans or other organisms reach harmful
proportions?

 

Present manufacture of contains about 0.02 parts TCDD per
million parts although manufacturers claim only about 0.05 ppm.
Some lots contain only about 0.01 ppm. A two pound per acre application
of will, therefore, distribute a little less than 20 ug TCDD/acre.
It is necessary to predict what may be expected to happen to that material
by examining existing laboratory data.

The environmental persistence of TCDD is highly dependent on the
conditions to which it is exposed. When mixed into soil, degradation
was found to be very slow, with a half time of one year (Kearney, 1972).
Because the material was introduced as an acetone solution there has
been some question about possible and limited access of
TCDD to soil organisms. However, TCDD in deposited on leaves and
exposed to direct sunlight was found to degrade with a half-time of a
little more than an hour (Crosby et al., 1977). The latter study shed a
good deal of light on earlier studies in which pure TCDD was carefully
coated on clean glass slides, water, or dry sterile soil for exposure to
sunlight, and found to be a1m05t insensitive to phOtodecomposition (Plimmer
et al., 1973; Kearney et al., 1972). Crosby et al. noted that reduction
would proceed in methanol, and that the reaction slowed significantly in
highly purified reagent. The finding that benzene accelerated the degra-
dation of TCDD in aqueous systems (Plimmer et al., 1973) suggested that a
good hydrogen donor was essential to photodecomposition. This conclusion
has been verified in more recent work (Crosby et al., 1977). In that study
TCDD with was also applied gn_soil, and a half-time of 50-55 hours
could be inferred from the data. This finding provides some suggestion
about the reaction time in indirect light, because the rough surface will
provide shaded areas where reaction must be slower. As yet, studies of
the effect of various parts of the spectrum emerging from light trans-
mitted through leaves, or reflected from various surfaces has not been
done. The absorption maximum for TCDD is known (Crosby et al., 1973)
but the relative sensitivity to degradation at that wave length apparently
is not.

Accumulation of TCDD in the biota has been studied both in the labo-
ratory and in the field, but there are still many gaps in the needed data.

In spite of the limited solubility of TCDD (0.2 ppb), it can accumu-
late in aquatic organisms to the extent it is available on sediments or
other reservoir (Matsumura and Benezet, 1973). In their experiments,
TCDD was deposited on sand, in solvent which was evaporated to leave the
dioxin as a film on sand particles. The sand was placed in a small con-
fined static aqueous system and various aquatic organisms added. Brine
shrimp reached a concentration of 157 ppb, mosquito larvae concentrated
TCDD to 4150 and silverside fish accumulated virtually no TCDD. It
may be assumed that the concentration of TCDD in water was maintained
throughout the experiment by the excess dioxin residue on sand.

18%?

15

 

When sufficient TCDD to represent 162 in the aquarium was ingested
by algae which were then placed in an aquarium with Daphnia and with Os-
tracods, the latter species accumulated TCDD to concentrations of 879 and
279 ppb, respectively. They also showed that mosquite fish concentrate
TCDD to a lesser extent than do the bottom feeding larvae on which they
feed. It is clear that accumulation does occur, but depends on maintaining
saturation of the ambient water.

Matsumura and Benezet (1975) also examined movement of TCDD which had
been bound to sand, to a sandy loam soil (inorganic to organic soil). Very
little TCDD was-translocated by a slow leaching with water. Subsequently,
a more complex food chain system of soil, water, algae, duckweed, snails,
daphnids, gambusia, and catfish was assembled (Isensee and Jones, 1975).
14C-TCDD bound to soil in concentrations varying from 0.0001 to 7.45 
was placed in aquaria. The residues found in the various species after
33 days exposure tended to peak at the Daphnia Stage. Gambusia were added
at the end of the exposure period with access to water and other organisms
for 3 days. Catfish fingerlings were exposed for 6 days, beginning after
all Other organisms were removed. The catfish accumulated TCDD to roughly
the concentration found in the first biological stage, the algae. None-
theless, at a water concentration of 7 TCDD a terminal accumulation
ratio of more than 10,000 was found in catfish, representing TCDD con-
centrations of about 100 (Isensee and Jones, 1975). The work also
shows, however, that as soil or water concentrations decrease, so also
do concentrations in the organisms of the system. Toxicity was not studied
although the higher tissue accumulations exceeded mammalian toxic doses,
assuming uniform distribution of dosage. The authors point out that
with the long latency of TCDD toxicity, the fish may not have had time
to develop lesions and die.

Ward (1976) examined the Persistence of TCDD in lake water and sedi-
ments. As expected, TCDD is almost entirely bound to lake sediments
and is subject to very limited but nonetheless real metabolic degradation
when microorganisms were present. The slow microbial activity is enhanced
by presence of nutrients. Ward suggested that water-mediated evaporation
of TLDD occurred to a limited extent.

Toxicity of TCDD to coho salmon has been assayed by Miller et a1.
(1973). TCDD was administered in the diet and dosage expressed as ng
TCDD per gram organism mass. At 13.1 ng/g only about 30% of the experi-
mental group survived for 80 days past exposure, and at 5.4 ng/g almOSt
half the fish did not survive. Deaths often did not occur until 10 days
after the end of exposure, and some lethally affected fish survived 60
days or more. In studies of rainbow trout there were no apparent effects
at intakes of 6.3 ng or greater. Mosquito larvae pupated at 3 nor-
mal rate in water containing 200 snails reproduced normally
and oligochaete worms suffered some reproductive deficit at the same
concentration.- Beatty et al. (1976) found Rana catesbiana tadpoles to
be remarkably resistant, with no apparent lethal effect by intraperi-
toneal doses of 1000 pg Doses up to 500 ug/kg did not affect
adult frogs, and no histopathological lesions appeared.

16438

16

Field studies of fish and mammals have provided evidence both sup-
porting and contradicting claims of TCDD accumulation in higher animals.
An evaluation of biota in the area used by USAF for training bomber
crews in application of military defoliants showed little accumulation
in fish living in the streams of the area. The area was liter-
ally saturated with receiving on the order of 1000 lb/acre during
the peak year, containing up to 2-10 TCDD. Mice in the area accumu-
lated tissue burdens of several hundred as measured at the end of the
spray program, when the sandy soil had accumulated residues up to 700
ppt. _These animals demonstrated no pathological changes (Young et al.,
1974), which is not surprising when extrapolating the data of Rose et al.
(1976) and Kociba et a1. (1976) to relate tissue levels and effective dose
rates in rats. Given the extended period of exposure, it seems possible
that the mouse population could adapt through survival of non-reSponding
strains over the estimated 30 generations of exposures. It may be also
that fish strains have evolved similarly as non-accumulators of TCDD, al-
though such adaptation seems unlikely.

 

Fish have also been analyzed in drainages from areas in Texas and
Arkansas which had been extensively treated with (Shadoff et al.,
1977). The Texas samples were from a pond which collected run-off from
a watershed on which the herbicide was used for brush control. The
Arkansas site was a pond into which adjacent rice fields were drained
and from which irrigation water was obtained, in essence recycling any
contaminants. Treatment was 1.25 lb 4-8 weeks prior to
flooding. The cycle had been in use for 18 years. Fish from the Texas
site did not contain TCDD, at detection limits of 5-7 ppt. Mud at a
detection limit of 3 and water at 0.1 were also negative. Six
human milk samples obtained from the San Angelo area near the Texas site
were also negative at a detection limit of 3 ppt. No TCDD was found in
the Arkansas fish samples. Meselson and O'Keefe [correspondence to Rep-
resentative James Weaver, 1977) have presented preliminary findings of
about 1-2 TCDD in human milk from the San Angelo area, and from an area
in western Oregon. 

Cows milk samples from areas of Oklahoma, Arkansas,
and Missouri were fOund to be negative for TCDD by Mahle et a1. (1977)
with detection limits on the order of ppt. TCDD in beef fat was un-
detectable in two series of animals which had grazed on treated
pastures in Oklahoma, Texas, and Missouri. In a third group of seVen
animals confined in an entirely sprayed pasture, three samples were posi-
tive at the detection limit of 3-4 (Kocher et al., 1977). TCDD has
been found, however, in several beef cattle maintained in a field experi-
ment on fields which had received 1, 2, 3, or 4 Average
TCDD in fat was 20.3 in the 4 lb/acre group, but there were only three
samples, one of which was extremely high. At 3 lb/acre the average was
12.8 ppt, again derived from some high values and many negatives. Inter-
pretation of this data by EPA is still in process, and is subject to some
argument.

A number of wild animal samples were obtained in treated forest areas
of western Oregon, several of which contained unusually high concentrations
of TCDD. The unique character of the data raises some questions, Since



17

 

1.1- 80



DOWthere seems to be few or no samples containing TCDD concentrations in-
termediate between the low background level and the 100-200 values in
a few animals. 

An issue has been raised about formation of TCDD during combustion
of and subsequent diStribution in air. Stehl and Lamparski (1977)
ignited grass treated with in a system where combustion was self
supported and found less than 0.0002% converted to TCDD. Combustion of
and trichlorophenol on filter paper led to production of even less.
TCDD. If the conversion of a field application were at this rate, about 2
parts of available per million would be converted, which is con-
siderably more than the residual contaminant level. It should be remem-
bered, however, that burning would not be done until foliage dries, and
with a 1-2 week half time it is doubtfu1 whether much herbicide would
be available for conversion. Even this maximum level of conversion is
probably not of great concern, since the product would be diluted with
combustion gases and air in millions of cubic meters of atmosphere. The
effect of exposure to light on decomposition while in the atmosphere is
unknown. Masking or wavelength shifts may have an inhibitory influence on
photodegradation, but the duration of atmospheric suSpension and light
exposure should enhance the reaction.

Induction of Microsomal Enzymes

 

TCDD has been found to be a remarkably potent inducer of many enzymes
which act on foreign chemicals. The effect is apparently dependent upon
of additional enzyme, rather than activation of partially syn-
thesized or otherwise inactive complete protein. TCDD also increases
activity of 6-aminolevulinic acid in some species, resulting in
an accumulation of in tissues and excretory routes. The potency
of TCDD for inducing some enzymes is between four and five orders of mag-
nitude greater than that of such classical inducing agents as 3-methyl
cholanthrene or phenobarbital. According to Hook et al. (1975a) TCDD
induces cytochrome P-450 at doses of fewer molecules of TCDD than there
are of P-4SO in the liver. This property is important because the
induced enzymes may increase conversion of other organic intoxicants
to non-toxic products, or conversely may increase conversion of non-toxic
materials to highly reactive intermediates which exert profound toxic
effects. Many carcinogens are activated in this way. The changes in
"drug?metabolizing" enzymes also have been used to describe profound
genetic differences among animal strains, and their respective response
to intoxication.

Enzyme induction may also serve as a predictor of general toxicity,
since the induction pctency seems well correlated with lethal, acnegenic,
and teratologic potential (Schwetz et al., 1973; Poland and Glover, 1973a).

The enzymes which oxidize foreign chemicals have evolved in response
to reactive groupings on molecules found in nature. While natural and
chemicals may represent an almost infinite variety of struc-
tures, the building blocks are the same, and they exist in relatively
limited numbers. Consequently, organisms have not had to develop an
infinite number of enzymes to survive, but rather have developed en-
zymes Specific for reactive groups.

18

The effeCts on these systems are among the more sensitive changes
found to be caused by TCDD. To simplify presentation the following table
identifies specific enzymes and the TCDD doses at which responses have
been observed. Unfortunately, few explorations of the entire dose res?
ponse curve have been made; the "no observed effect" intake has usually
not been determined, nor has there been an adequate study of the impact of
low level chronic studies on the microsomal enzymes. Most of the data in
the table refers to rats. Relatively few studies of mice have been made,
and in the guinea pig the effective doses approach the lethal dose, and
observed changes are minor. Hook et al. (1975b) administered 0.175 ug/kg
to guinea pigs and found a slight increase in liver biphenyl-4-hydroxylase
and biphenyl-Z-hydroxylase, and no change in aryl hydrocarbon hydroxylase
or UDP glucuronyl transferase.



 

Several generalizations appear applicable to the enzyme inductive
prOperties of TCDD. While most microsomal enzymes are increased after
TCDD, the enzymes identified as demethylating aminOpyrine, benzPhetamine
and morphine are decreased (Lucier et al., 1973).

Females seem to be more sensitive to the inducing effects of TCDD
(Lucier et al., 1975b). While effects on UDP glucuronyl transferases
are substantial, steroid glucuronyl transferases are not changed. The
bulk of induction occurs in liver but a similar but usually lesser in-
crease can occur in the kidney and some other tissues (see table). In
kidney, activity seems concentrated in the "outer stripe? of the medulla
and in the cortex. This region contains the terminal Straight segments
of the proximal tubules, which appear to be the only significantly in-
ducible cells in the kidney (Fowler et al., 1977).

The induction of aryl hydrocarbon hydroxylase (AHH) by TCDD has been
very useful in explaining genetic variability in receptor and effector
mechanisms. TCDD is about 3 104 times more potent than 
threne, producing half maximal stimulation in the rat liver at 0.85
nmoles/kg and in the mouse (female) at a dose of 0.42 nmoles TCDD
per kg (Poland and Glover, 1974). While the susceptibility to AHH induc-
tion is about the same in the species tested (ED5 0.4-1.2 nmole/kg), and
the LDsos differ by about two orders of magnitude, this difference does
not detract from the value of the system in studying the genetics of drug
response and the great problems of nan-homogeneity of exposed populations.

There is known to be a profound difference in the responses of various
strains of mice to AHH induction by (S-MC), leading
to a designation of "reSponsive" and "non-responsiVe" strains. Even at
high doses, S-MC fails to cause induction in the non-responsive animals.
TCDD, however, will elicit an amplified enzyme activity in such mice (Poland
and Glover, 1975; Chhabra et al., 1974).

Poland and Glover (1975) and Neubert et a1. (1975) have shown that the
genetically non?reSponsive mice have the genetic apparatus necessary
for expression of the inducible activities and suggest that the difference
arises from a mutant form of inducer-binding receptor with a diminished
affinity for aromatic hydrocarbons. The induction process is apparently
an increase in de novo of enzyme, rather than activation of
preexisting protEin or decreased degradation of protein (Haugen et al.,

 

he
on
e213
#5


19

nan
?ux
RU
?i

Induction of Foreign Campound Metabolizing Enzymes by TCDD 

iae?sri?g

306 

 

 

 

OZ

Extent of .
Initial Duration of Remarks (liver 5
TCDD Dose Induction Increased unless other 
Enzyme (pg/kg) (x normal) Activity References tissue specified)
p-Nitrophenol?UDP glucuronyl 3 14 >34d Lucier et al., 1975a pregnant rats (2)
transferase 3 7 >34d Lucier et al., 1975a neonate, (3)
2S 6 73d Lucier et al., 1975b male rat
5 >28d Hook et a1., 1975b
0.2 2.5 -- Lucier et 31., 1973
Ben2pyrene (aryl hydrocarbon) 3 l4 >21d Lucier et al., 1975a pregnant rats (2)
hydroxylase 3 2 Lucier et al., 1975a neonate, (4)
2.5 14 -- Berry et al., 1976 pregnant rat, (5)
2.5 100 -- Berry et al., 1976 fetal liver
0.2 7 -- Lucier, 1973 male rats (6)
0.2 8 -- Hook et a1., 1975b female rats (7)
1.0 1.5 -- Hook et al., 1975b male (7)
25 3 -- Hook et al., 1975b male rat liver
25 >70 -- Hook et al., 1975b male rat kidney (8)
10 180 -- Poland and Clover, 1974 rat kidney
10 10 Poland and Glover, 1974 rat lung
10 100 -- Poland and Glover, 1974 rat intestine
.08 3 -- Poland and Clover, 1974 mouse
NADPH diaphorase (dehydrogenase) 90 13 >15d Beatty and Neal, 1976 male rat liver (9)
Aniline hydroxylase 5 2 >15d Lucier et al., 1973
0.2 1.2 -- Lucier et a1., 1973
Cytochrome 1.0 1.6 Lucier et a1., 1973 male, 3 day
Cytochrome B5 1.0 1.4 -- Lucier, 1977 male, 3 day
Biphenyl-Z-hydroxylase 25 13 -- Hook et 31., 1975b rat liver
25 18 Hook et al., 1975b rat kidney (11)

geesi

IZ

Induction of Foreign Compound Metabolizing Enzymes by TCDD (cont)

References

Remarks (liver
Unless other
tissue Specified)

 

 

 

Extent of
Initial Duration of

TCDD Dose Induction Increased

Enzyme (Hg/kg) (x normal] Activity
Biphenyl-4~hydroxylase 25 2
25 4O

Amino levulinic acid 25 none --

25 2
1.5 ng/egg 2

Hook et al., 1975b
Hook et al., 1975b

Woods, 1973

Goldstein, 1973

Poland and Glover,
1973c

rat liver
rat kidney

mouse (11)
chick embryo

 

1This table is selective and is intended to show the variety of work done, the dose ranges studied, and duration

of effects.
The references noted should be consulted for greater detail.

2Pregnant rats, treated on day 10 of 23 day gestation, sacrificed 21 days post partum.

3Mother treated at day of gestation; activity measured at day 21 postnatal.

parturition.

Same as 2 except measured at day 8 postnatal.

(HA

TCDD at day 17 of gestation, sacrificed at day 20.

0?

Male rats, sacrificed 3 days after TCDD.



In most cases the lowest dose producing effect has been shown and data for higher doses omitted.

No increase in activity before

Male rats did not reSpond at this dose; there was a small but significant (P 0.05) increase in amino pyrine

demethylase and cytochrome P-450 in females at 0.2 pg 

8Resting activity below detectiOn limit.

9Maximal activity in cytosol at 7 days, still rising in microsomes at 15 days.

10
11

ug/kg produced no effect.

Resting activity very low in these tissues; factor of increase approximate.

we 

 

1976). Whether the effect of TCDD on inducible enzymes has implications
on other aspects of protein can only be speculated upon.
Poland and Glover (1975) showed about a 10?fold difference in the dose
necessary to cause induction. Through the finding that heterozygous
of (responsive) and (non-reSponsive) strains
are intermediate in sensitivity, they support the contention of Poland
and Glover (1975) that the difference is a receptor site mutation. Later
studies have strengthened that premise in finding that hepatic accumula?
tion of 14C-TCDD administered intraperitoneally was greater in respon-
sive than non-responsive strains (Poland et al., 1976). In the same
study, in_vitro examination of binding in the soluble fraction of liVer
cells showed that specific binding sites exist, and that their affinity
correlates with the genetic capability for induction. Furthermore,
tests of an extended series of halogenated dioxins and dibenzofurans
showed that cytosolic binding corresponded closely with the induction
potency of the chemical.

Niwa et a1. (1975) have measured AHH induction by TCDD in a variety
of cell cultures. They used 10 e5tablished cell lines, human 
cytes, and primary fetal cultures from chick, rat, rabbit, hamster, and
four strains of mice. Generally, kinetics of TCDD induction in each
cell type is similar to that of S-MC, and the induCtion was inhibited
by actinomycin-D and cyclohexamide, which are inhibitors of protein syn-
thesis. They found no relation between inducibility and cytotoxicity
of TCDD, which is further evidence that TCDD itself is not metabolized.
The sensitivity of some cell lines is such that the authors suggest use
of H-4-II-E cells (derived from.Reuber hepatoma H-SS in rats) as a bio-
aSSay, with a suggested sensitivity below 1 pMole (N 0.0003 pg) in 3 ml
of culture medium.

The same kind of genetic differences have been studied in human
cultures, which represent the genetic background of the cell
donor and are therefore useful in making relatively non-invasive studies
of potential human response to intoxication. The sensitivity to TCDD
induction of AHH is about 50-fold greater than to S-MC induction, con-
siderably less than the 30,000-fold difference in responsive mice (Kouri
et al., 1974). Atlas et a1. (1976) have carried these studies forward
with from human tissues, but have as yet dealt only with S-MC,
not TCDD. -Part of the stimulus for examining human cells lies in the sug-
gestion that extent of induction at contact sites skin, lung) may
relate to the probability of cancer initiation by activated carcinogens
(Kellerman et al., 1973). Although TCDD is as yet not established as
a carcinogen such research may provide suggestions about the extent of
expected variations in other TCDD responses in humans.



General Toxicity in Laboratory Animals

 

Apparently the first published account of acute toxicity
was that of Drill and Hiratzka (1953). The L050 for dogs was about 100
mg/kg; the principal was a mild incoordination. When fed five
days weekly for 90 days, doses up to 10 mg/kg/daily were without evident

15445

22

effect, but 20 mg/kg/day was lethal to all 4 treated animals between 11
and 75 days after the first dose. were limited to muscle
twitching and impaired swallowing. A field study by Grigsby and Farwell
(1950) [as quoted by Rowe and Hymas (1954)] exposed a variety of stock

on pasture immediately after spraying at 2?4 times usual levels,
with no effect.

 

6091100 MOO

Rowe and Hymas (1954) summarized the available data at that time
and estimated the acute oral median lethal dose for male rats to be about
500 mg/kg, male mice 389 mg/kg, and guinea pigs 381 mg/kg. LDsos for
various esters were higher than for the acid. A sheep fed the propyl
glycol butyl ester of died after 369 daily doses of 100 mg/kg
and another sheep and a cow died after seven daily doses of 250 mg/kg
(Palmer and Radeleff, 1964). The triethylamine salt of at 100 mg/kg
caused no observable effect after 481 days of treatment at 100 mg daily. As
single animal observations these can be considered rough estimates only,
but they convey the generally high doses necessary to cause harm in rumi-
nants. has been shown to cause decreased volatile fatty acid
production in vitro at concentrations of 500 ug/ml or greater (Kutches
et al., 19757} this concentration is probably greater than can be main?
tained by a survivable daily dose of 

Subchronic (90 day) feeding of male and female rats caused no ef-
fects below 30 mg/kg/day, but body weight and food intake were depressed
after treatment at 100 mg/kg/day. Alkaline phosphatase and SGPT were
elevated and count and hemoglobin was decreased. The his-
topathologic findings were limited and inconsistent (McCollister and
Kociba, 1970). Rats were able to tolerate 186 mg/kg/day treatment with
mixed mono-, di-, and tripropylene glycol butyl ether esters of 
over 90 days but developed some indications of toxicity. At a dose of

18.6 mg/kg no evidence of toxicity was observed (Dow Chemical Company,
1961).

A two year study of rats given 3 to 30 mg has very
recently been completed by Dow Chemical Company. Much of the analysis
has yet to be completed, including morphologic pathology. Animals that
received 30 mg were found to have increased urinary por-
phyrin excretion after 4 months of treatment, and this change continued
through the entire 2 year period. No changes were found in any other
hematologic, urinary, or clinical chemistry measurement at that rate of
intake, and the increased excretion did not occur at 10 or 3
mg/kg/day (R.J. Kociba, Dow Chemical Company, preliminary status report;
personal communication].

A feeding Study of reindeer was prompted by allegations that a high
incidence of death and abortion occurred in 1970 in a herd after use of
phenoxy herbicides the previous year. Fifteen of thirty pregnant rein?
deer were fed birch leaves which had been sprayed with a 
mixture, through a 1.5 month period late in gestation. The daily dose of
phenoxy acid was about 1 mg/kg/day. No clinical chemical changes could
be detected, nor did any changes appear at autopsy either in the adult
animal or the full term fetus (Erne, 1976).

reass-

23

 

The toxic effects of in rodents have been studied in much
more detail than the gross toxicity studies already described. Highman
et al. (1976a, 1976b) established that lethal doses in pregnant or nonpreg-
nant mice caused myocardial lesions, bone marrow aplasia, and 
depletion in thumus, spleen, and nodes, and that animals which re-
mained apparently healthy despite similar doses did not suffer the same
severity of lesions. A mild hemolytic anemia did occur in such animals.
Pregnancy did not augment the toxic effect. It was also clear that major
differences exist in the reSponse of specific strains. Many NCTR mice were
seriously affected at doses below 60 mg (6-9 days of treat-
ment while most CRBL mice remained unaffected at doses as high as 90-120
mg/kg/day at the same time. The histopathology, hematoIOgy, and blood
chemistry changes in treated mice were also Studied. In many monitored
animals, some myocardial fibers were found to be pale and swollen, with
longitudinal striations. The condition was rarely seen in treated ap-
parently non-intoxicated animals. Necrotic changes were also often seen
in the outer myocardium. High doses caused thymic atrophy, with almost no
in the cortex and increased numbers in the medulla (this ef-
fect was enhanced in late pregnancy by the normal tendency to cortical
involution in pregnant mice). Spleens were often atrophied, and thyroid
fbllicles were enlarged with enlarged epithelial cell. In the liver,
glycogen was often depleted. Blood chemistry changes appeared not to be
marked.

has been found to cause increased liver weight in rats at
doses of 167-334 mg/kg over a two-day period, apparently through stimu-
lation of protein and RNA but not as newly formed 
metabolizing enzymes. The change reverses after withdrawal (Chang et al.,
1974; Rip and Cherry, 1976). Liver nuclei isolated from treated animals
were more active in RNA than those from controls. at
a concentration of 4 mmolar is capable of sharply inhibiting in_vitro in~
corporation of mevalonate-14C into non-saponifiable lipids by rat liVer
(Olson et al., 1974). If data obtained in mice by Nony et al. (1976) can
be applied, this concentration is approachable at single doses of 50 to
100 mg/kg, which are survivable by most Species.

Koschier and Berndt (1976a, 1976b, 1976c) have described the effect
of upon renal physiology. Large doses of appear to
impair secretion of itself by decreasing the activity of the organic acid
transport system in the proximal tubule. Organic base tranSport was 
also depressed. Small doses of were excreted very rapidly, but
large daily doses led to renal depression and retention of the compound.
The authors consider the data to support the conclusion that transport
of phenoxy herbicides is an active process.

The of does not seem to be a popular
field of study. In the one study fOund, single doses of up to 100 mg/kg
were given pregnancy. The male of females
given the highest dose exhibited more exploratory open field behavior,
but no difference was found in females. The highest doses caused de-
creased litter size but no increase in malformations (Sjbd?n and deerberg,
1972).

15447

24

Poultry seem relatively insensitive to Whitehead and
Pettigrew (1972) found that a single oral dose of 900 mg/kg to 4 week old
chicks caused 40% lethality. However, feeding of 1000 mg per kg
diet/day for three weeks to chicks, beginning at one day of age caused
only some slowing of growth; 5000 mg/kg diet was lethal. At levels not
causing gross toxicity, plasma calcium and magnesium were not affected.
Given a choice, the birds rejected the treated diet in favor of non-
contaminated food. Turkeys fed at a rate equivalent to 62 mg
acid daily for 11 days were unaffected (Roberts and Rogers, 1957).
Bjorklund and Erne (1971) introduced various derivatives into
water and feed of chickens, quail, pheasants, and ducks. In water the
LCloo of the triethanolamine salt for chickens over a 29 week period was
1000 acid equiv. [about 200 mg/kg). In the diet of other Species over
a 7-day period, the LC50 was in excess of 2000 ppm. Kenaga (1975) has
extensively reviewed avian toxicity and safety of birds in areas treated
with herbicides and concludes that the no observed effect levels are
substantially above amounts that might be contacted in a field application.

 

Effects of on Reproduction

 

The teratogenic potential of has received wide publicity
since 1969 when allegations were made that its use as a military defoli?
ant had caused fetal malformations in the Vietnamese population. A
succession of studies over the next three years confirmed the terato-
genicity of but the implication of the herbicide in any increase
in birth defects has net been supported. The initial report of 
teratogenic effect (Courtney et a1., 1970; Bionetics Research Laboratories,
1970) described cleft palate and cystic kidneys at doses of 46 and 113
mg/kg/day on days 6 through 14 of gestation. 0f the two lesions, cystic
kidney appeared to be a more sensitive indicator. The used in the
study was found to contain 30 
(TCDD), and the respective toxicities of the two agents were still unclear
at that time. The data was also criticized for an unusually high and
variable incidence of embryotoxicity in control animals (Neubert and
Dillman, 1972).

Emerson et a1. (1971) evaluated a commercial with less than
0.5 TCDD and found that 24 and 40 mg/kg/day through days 6-15 of
gestation caused no teratogenesis in rats. Sparschu et a1. (1971) then
found that 50 mg was also nonteratOgenic, although there was a slight
increase in incidence of delayed skull ossification. (Such alteration in
deVelopment is not considered teratologic, because the abnormality dis-
appears with age. One criterion of teratogenic effect is irreversibility.)
Treatment at 100 mg/day for days 6-10 caused generalized maternal toxi-
city, with only 4 survivors of 25 treated animals. Intestinal hemorrhage,
considered a common fetotoxic but not teratologic lesion, was found in
only one pup in this study. Khera and McKinley (1972) found a similar
dose response.

In mice the effective dose is similar to that in rats. At 50 mg
through days 6-15 of gestation, the frequency of cleft
palates was increased from 4.7% to 20% and resorption frequency increased.
The incidence was raised to 73% by a dose of 110 mg/kg/day with a decrease

25

W. "at:



00.1.128le

in fetal weight (Bage et al., 1973). The high dose also caused a two?fold
increase in rib and vertebral malformation, but only marginal increase

in dilated renal pelvis. Neubert and Dillman (1972) found increased
cleft palate in NMRI mice at doses over 200 mg/kg/day through days 6-15,
increased embryo lethality at doses over 45 mg/kg/day reduction in fetal
weight above 15 mg/kg/day. Strain differences in mice are significant.
The dose causing at least one cleft palate in 50% of litters was 25 mg/
kg/day in the most sensitive of 5 strains tested and 105 mg/kg/day in the
least sensitive (Gaines et al., 1975). Roll (1971) found detectable
teratogenesis at doses above 35 mg/kg/day through days 6-15 of pregnancy.
The no?effect level with respect to teratogenesis was established as 20
mg/kg/day.

Hamsrers are less sensitive to cleft palate, but do have a somewhat
higher incidence of delayed head ossification which is not generally
defined as a teratologic manifestation (Collins and Williams, 1971).
doses ranged from 40-100 mg/kg and there was a considerable difference
among Samples of of different sources and levels of TCDD contamie
nation.

The

Fetal rabbits are apparently unaffected by maternal doses up.to 40
mg through days 6-18.

Ruminants are apparently also quite resistant to teratogenic effects
of Binns and Balls (1971) fed 100 mg to 11 ewes from
the 14th to 36th day of gestation, and fed 100 mg to
another group during the same period. No evidence of deformity appeared
in any of the lambs. The TCDD content was 1 ppm.

The no apparent effect levels in rodents reported in the proceeding
studies contrast sharply with data in abstracts of reports by Konstantinova
(1974a, 1974b), who is said to have found that 4.2 mg/kg/day of the butyl
ester of through the entire gestation period caused embryotoxicity,
nervous and hematologic change and histopathology in the mother. A dose
rate of 0.42 mg/kg affected growth and development, nervous, liver and
kidney functions and lowered fertility. The succeeding generation was
found to have minor changes in organ weights. (Direct translations of
these papers are not available; data quoted is obtained from Chemical
Abstracts.)

There has been limited study of the teratogenic effect of 
in primates. Daugherty et al. (1975) adminisrered 0.5, 1.0, and 10 mg/
kg/day to groups of 10 pregnant rhesus monkeys from day 22 through 38 of
gestation. The high dose was established after finding that 12 mg or
more daily for 18 days caused vomiting and weight loss in
monkeys of both sexes. No teratogenesis occurred, although there were
1 or 2 abortions, premature births, or neonatal deaths in all groups,
including the control.

The importance of TCDD in the teratogenicity of was examined

soon after the contamination was recognized. Courtney and Moore (1971)
tested with 0.5 and 0.05 TCDD, and TCDD together and

1844.9

26

MOO

TCDD alone in mice. It was evident that the concentrations used or addi-
tion of 1 ug TCDD/kg/day through days 6-15 did not alter the apparent
teratogenicity of Neubert and Dillman (1972) reached a similar
conclusion, suggesting that to potentiate effect on pregnant mice
at least 1.5 TCDD is necessary. Collins and Williams (1971) observed
additive effects of 2.9 and 45 TCDD contamination of adminis-
tered to hamsters, but many of the changes appeared characteristic of pri-
mary TCDD intoxication.

-



919$:

 

Nutritional status seems to have little influence on the effect of
on reproductive Hall [1972) fed 250 and 1000 
with diets containing 20% and 60% casein. Resorption and still-
birth incidence and litter size were similar in all groups, although fetal
organ and placenta weights were decreased in the high high pro-
tein group. Effects of low protein intake were augmented by the high
concentration of herbicide.

Incubating eggs have been considered to be particularly vulnerable to
herbicides because they remain in one place, and because the eggshell is
porous. A number of studies of effect on eggs have appeared since
1967, with conflicting results (see section on but has not
received much attention. Somers et a1. (1973) sprayed hen eggs with a
formulation containing and 2,4-0 in a treatment equivalent to 11.2
kg/hectare (10 times normal field application rates) prior to incubation
without effect on hatchability or early survivability of chicks. They then
treated eggs of the pheasant, as a genetically more heterogeneous species,
in the same way, without adverse effect. Entry of herbicide into the egg
was verified analytically. The treatment did, however, increase weight
gain in male chicks during the first four weeks of life (Somers et al.,
1974). The L050 of in DMSO, injected directly into the air space
of chicken eggs at doses up to 125 mg/kg has been calculated acetone carrier, 133 mg/kg (Strange et al., 1976). No tera-
togenic effect was evident. The solvent toxicity was shown to be substan-
tial, but it seems clear that acquisition of an effective dose of 
by an egg in the field is highly unlikely.

In fact, immersion of hen eggs in 1% was ineffective, and a
solution was only moderately effective. The eggs were immersed for 10
seconds, then returned to incubation (Gyrd-Hansen and Dalgaard-Mikkelsen,
1974).

Insects may be more sensitive; Davring and Sunner (1971) and D?vring
(1975) have shown that while DrOSOphila are highly resistant to lethal
effects of ester (L050 4700 in the diet), 1 caused
disturbed egg follicle development and chromosomal defects in developed
oocytes.

A study on a species of killifish has shown that concentrations of
20 have substantial teratogenic effect. The embryos developed several
cardiovascular anomalies, and occasional and splenic defects. The 20
treatment reduced hatchability to less than 50%, and 25 allowed
less than 5% to hatch. No anomalies were found after treatment with 14
and the hatch was reduced only 4% (Schreiweis and Murray,
1976).



27

 

Carcinogenic and Mutagenic Potential of 

Probably the most important issue about any chemical introduced into
the environment by human activity is the possibility that it may increase
the incidence of cancer in the human population. The probability of muta-
genic activity is of almost equivalent concern, both because of the possi-
bility of genetic alteration and because mutagenesis may be a useful pre-
dictor of carcinogenic activity. The relationship is by no means constant.
However, McCann et al. (1975) have assembled data from a number of labo-
ratories using microbial systems ("Ames test") and find that 85% of known
carcinogens are positive, 10% of non-carcinogens are active. This is not
to say that almost every agent identified as a mutagen will be carcinogenic,
but the test should become an aid in deciding priorities for direct screen?
ing.

Only a limited number of experimental studies of carcinogeni-
city have been made. A screening program for 120 pesticides and industrial
chemicals was reported by Innes et a1. (1969). Eleven of the agents caused
an elevated incidence of tumors; none of the phenoxy herbicides, including
caused increased tumor formation at a dose previously determined to
be the maximum tolerable daily dose over 20 days, without lethality (21.5
mg/kg/day for Muranyi-Kovacs et a1. (1976) treated two strains of
mice with 80 (TCDD content 0.05 ppm) in the diet for more than
500 days. Daily dosage was about 12?15 mg/kg. In one strain survival time
was significantly decreased in males, in the other significantly increased
in females. An apparently significant small increase in tumor incidence
occurred with the increase in life span. The authors are concerned that
altered life Span may confound the analysis, but consider that 
can not yet confidently be considered non-carcinogenic, and requires fur-
ther analysis.

A two year study of rats given up to 30 mg has just
been terminated by Dow Chemical Company, but the pathological evaluation
had not been completed at the time of this preparation.

One epidemiological Study has been made of tumor incidence in a
group of Swedish railroad employees. It was possible to isolate groups
who had handled phenoxy acids exclusively, and a cohort of 207 persons
with at least 45 days of exposure, totaling 1747 person-years. No in-
crease in tumor incidence could be detected in these workers although
increased incidence appeared in groups exposed to other herbicides
(Axelson and Sundell, 1974).

Mutagenic assessments have been carried out on a number of systems.
Buselmaier et a1. (1973) were not able to show induced mutations
in Salmonella typhimurium G46 his-, and Serratio marcescens a21 Leu- and
a31 his- in a host mediated assay in mice. Serum from animals treated
orally with also did not induce mutants in S. typhimurium his-
(Styles, 1973). Anderson et a1. (1972) were unable to demonstrate any
mutagenic preperties of or other phenoxy herbicides when tested
against eight histidine requiring mutants of S. typhimirium.

i'??Si

28

Sex linked lethal tests of adult male Drosophila showed to
be negative, although fertility was decreased (Vogel and Chandler, 1974).
Majumdar and Golia (1974) were able to show an increase in sex linked
recessive lethal mutations after 15 day feeding on 1000 
non?significant increase was apparent after feeding 250 ppm.

1,360, Moo



A



918*

In gerbils six-day feeding of up to 150 mg produced no
increase in chlorosomal abnormalities, but higher doses did cause some
increase (Majumdar and Hall, 1973). Jenssen and Renberg (1976) tested
for mutagenicity by seeking micronuclei in of mouse
bone marrow, which is a high resolution detection system. The system
was cytogenetically negative, although some mitotic depression was de-
tected. It was noted that only about 5% of circulating found its
way into the cells, which was suggested as indicating a limited hazard 
due to lack of access.

A limited survey of pesticide applicators who had been in contact
with a variety of agents showed some increase in chromatid gaps and breaks
during the spray season (Yoder et al., 1975). Recovery in the off season
reduced frequency below that of controls, suggesting an augmented repair
system. was the least frequently used agent, and it is not
likely that any observed effect can be attributed to it.

Dow Chemical Company has maintained a program of continuing health
monitoring of production workers. Cytogenetic evaluations of
52 men in early 1970 were reported to be negative (Johnson, 1971), and
a second analysis in 1974 also found no evidence of abnormality (Kilian,
1975). An interesting finding, however, was that the fraction of each
exposed group found to have no abnormal cells was above 80%, while of the
control group only 74% had no abnormal cells.

Absorption, Distribution, Metabolism, and Excretion of 2,4,911

has a rather short residence time in all species. A cow
given 450 mg of acid in four daily doses excreted the entire
amount in urine in six days (St. John et a1., 1964). Erne [1966a, 1966b)
administered 100 mg of the amine salt/kg to rats and swine and found
plasma half time for the rat to be three hours, and 10 hours for pigs.
Kidney, liver, lungs, and spleen concentrations of could occa?
sionally be forced above plasma levels, but apparently none of the
material entered the brain or adipose tissue. Tissue half times ranged
from 5-30 hours. When the agent was administered repeatedly plasma levels
tended to lessen, with increased excretion. Of the in blood, about
20% was in is excreted more slowly by mice than by
rats, at a rate of 1-4% of the original dose per hour (Zielinski and
Fishbein, 1967).

By overloading sheep (four 250 mg doses) it is possible to produce
residues in tissues. Maximum accumulations were abOut 100 in fat and
muscle. In each case residues were found in the acid form regardless of
the form in Which it was fed (Clark et al., 1970). The latter finding is
at some variance with Clark et a1. (1971), in which the PGBE ester of 
was reported to remain as the ester in urine and tissues. Cattle fed 0.15

Ci??




29

 

and 0.75 mg proPylene glycol or butyl ether esters/kg/day for 32
weeks, produced no increase over background residue. Clark et a1. (1971)
later reported feeding sheep 2000 in the diet and finding 1.0
in muscle. No residue could be detected 7 days later. Deer
browsing on land treated for reforestation were found not to contain sig-
nificant tissue residues; measurable was found in stomach contents,
urine, and feces up to 43 days after herbicide application (Newton and
Norris, 1968). The implication, again, is that suStained field intake

will not cause detectable tissue residues.

Grunow et a1. (1971) found about 50-70% of administered 
in urine, and found that a substantial fraction emerged as derivatives,
one of which was identified as acetyl) glycine.
The taurine conjugate and 2,4,5-trichlorophenol have also been identified
(Grunow and Bohme, 1974). Nony et a1. (1976) have measured its
glycine amide and alkaline hydrolyzable conjugates in blood, urine, and
feces. The proportion of metabolites is quite low in blood, and increases
substantially in urine from 24-72 hours after administration of the herbi-
cide. In the feces the glycine amide is found in only limited amounts, but
other conjugates may account for as much as 25% of the total recovery.

Research with labeled in rats showed that the half-time
of plasma clearance was about 4.5 hours and essentially dose inde-
pendent below doses of 50 mg/kg but increased as dosage increased to 19.4
at 100 mg/kg and 25.2 hr at 200 mg/kg. The of a 5 mg/kg dose in
dogs was much longer, 77 hours. About 90% of the in blood was
reversibly bound to plasma prOtein, over a wide range of concentrations. It
was found also that as the dose was elevated, a small amount of fecal ex-
cretion took place. Detecrable, minute amounts of emerged in the res-
piratory gases (Piper et al., 1973). Fang et a1. (1973) also showed a
non-dose dependent at lower doses. Nony et a1. (1976) found in excess
of 10% of the toral excreted material in feces. The extent of prOtein
binding has been further defined by Kolberg et a1. (1973); bovine serum
albumin binds 13-14 moles BSA.

As suggested by the slower the percentage of dose excreted/day
decreased sharply at the higher doses, indicating a limit in excretory
capacity by the kidney. Rat kidney tissue slices can concentrate 
about 15 times, slices from dogs can concentrate about 9 times. Berndt
and Koschier (1973) found an energy dependent uptake of phenoxy herbicides
by renal cortical tissue. Metabolic inhibitors depress the process.
Acetate and lactate as substrates enhance the accumulation. Increased
in the medium slows the concentrating capacity of the kidney.

The above work was later carried forward by resorting to intravenous
injection of in order to take advantage of usefu1 pharmacokinetic
models (Sauerhoff et al., 1976). Doses of 5 mg/kg and 100 mg/kg were used,
and samples were taken through 36 hours (5 mg) and 72 hours (100 mg).

After 5 mg/kg rats excreted about 50% of the body burden every 12 hours,

in spite of a 4.3 hour plasma further illustrating the concentrating
capacity of the kidney. At 100 mg/kg the plasma was 23.1 hours for the
first 36 hours, then was comparable to the animals given a lower dose.

inass_

30

"Moo

In humans (Gehring et al. (1973) found that a 5 mg/kg dose was ex? 
creted with a of 23 hours for both plasma and whole body clearance. 
Essentially all of the adminisrered material emerged in urine unchanged. 

The phenoxy acids are relatively strong organic acids, which is one 
of the reasons they are excreted in large part as the parent molecule. f?kzg
The possibility of hydrolysis by rumen flora or by tissue enzymatic pro- 
cesses was first considered by Wright et al. (1970) who found that after NJ

feeding of to sheep,
2,4,5-trichlorophenol residues could be detected in tissues. Clark et a1.
(1975) measured phenoxy acid and phenol residues in muscle, fat, liver, and
kidney of sheep fed 2000 for 28 days, and found 1.00, 0.27,
2.29, and 27.2 and 0.13, <0.05, 6.1, and 0.9 2,4,5?tri-
chlor0phenol, respectively. Seven days after withdrawal, residues
were well below 0.1 or below the detection limit of 0.05 ppm. The
phenol remained at significant levels in both liver (4.4 ppm) and kidney
(0.81 ppm).

According to Koschier and Berndt (1976c) the rate of excretion rises
almOSt immediately to about 80% of input in animals treated daily, then
continues to rise for 7-9 days until excretion approximately equals input,
where it remains through the duration of treatment. The rate of adminis-
tration was 20 mg/kg/day. As dose rate increased the initial percentage
excreted was less, but 7-9 days was still required to reach a rate equal
to input. An apparent over-shoot then took place for 5-6 days.

The markedly different rates of excretion by rats and dogs
was Studied in_vitro by Hook et al. (1974). They found that kidney of
both species actively accumulated by a saturable, oxygen dependent
process which was depressed by other organic anions. Rat kidney had a
greater dependence on potassium, and dog kidney increased accumulation
of in the presence of acetate. The capacity of kidney to trans-
port the standard test anion, para?amino hippuric acid (PAH), can be com-
petitively inhibited by indicating that both are transported by the
same mechanism. The authors concluded that the greater capacity for PAH
transport by rat tissues accounts for the difference in retention between
rats and dogs. Apparently lower renal excretion is also responsible for
the extended half?time of in new-born rats observed by Fang et al.
(1973); Hook et al. (1974) have found that kidneys of 10 day old rats are
much less efficient than adults. Subsequent work by the same group (Hook
et al., 1976) indicated that plasma binding in dogs was more tenacious than
in rats.

The high specificity of in causing cleft palate has been
investigated by Dencker (1976) as a diStribution phenomenon. The sensitive
period has been shown to be late in fetal development, during days 12-13
(Neubert and Dillman, 1972). Palatal closure occurs very late in organo-.
genesis and is complete at about day 15.

Dencker (1976) points out that no unusual uptake of 2,4,5wT in these
structures occurs, but overall uptake in the fetus is greatly increased
between days 11 and 18. It may well be that this increase, at a time when
formation of most other StruCtures have progressed far enough that they are
not sensitive, is responsible for cleft palate.

16454

31

4

It is possible for to reach milk if a sufficient level is
included in the diet. At 30 in the diet for two weeks, followed by
one week on untreated feed, no or trichlorophenol was found at
a detection level of 0.05 ppm; at 100 trichlorophenol was barely detec-
table. At 1000 and 0.16-0.27 trichlorophenol
was found in milk, and somewhat less was found in cream (Bjerke et al.,
1972). These figures can be put in the perSpective of forage consumption.
Residues on grass are on the order of 100-150 ppm/lb/acre (Morton et al.
1967) immediately after application. These amounts drop sharply to about
20% of initial levels during the initial two weeks post-application. This
factor, with the rapid excretion characteristic of make even limited
human exposure through dairy products or meat highly unlikely unless prod-
ucts are obtained immediately after heavy Spraying.

are 

 



M00. 0011"

DOW

Behavior of in the Environment and Effects on Submammalian Species

One factor in the hazard potential of an herbicide is determined by
its persiStence after application and its movement and reactions in soil,
water, or in plants.

Appearance of in forest streams has been shown to result only
if the herbicide was introduced directly to the stream, and diminished
from about 0.1 to less than 0.01 within one day after treatment
(Norris, 1967). Norris (1968) also found that heavy rains six months
after treatment did not move detectable into streams.

The herbicide is usually degraded on the forest floor quite rapidly.
Norris (1969) found that nearly 90% disappeared in two months, although
cold weather, sterile soil, and lack of moiSture may extend the period of
degradation over as much as 9 months. The trichlorophenol hydrolysis pro-
duct of degrades faster than the parent compound {Alexander and
Aleem, 1961). Evaluation of several soils from South-Vietnam indicates
that concentrations on the order of those expected after a typical forest
application should be virtually gone in seven weeks (Byast and Hance, 1975).
More than 20% of the radioactive carbon of labeled had become in-
corporated in the soil, but actual residues of herbicide were usually
about has also been found short lived in various Texas soil
by Bovey and Baur (1972). Altom and Strittke (1973) studied in
three Oklahoma soils and found half-times of 24, 14, and 21 days. The
shorter time was on grass lands, the others were forests.

Leaching tests show that bound in soil remained in the upper
6 inches of test columns even after application of 4.5 inches of water
(Wiese and Davis, 1964).

The various esters of 2,4,5eT are often intrinsically more toxic
than the parent acid, probably because they are more fat soluble. In
an aqueous system, however, they hydrolyze rapidly to the acid, and the
rate is enhanced by soil and microorganisms (Teasley and Williams, 1970,
as quoted by Kenaga, 1974). It seems reasonable to expect a fairly rapid
hydrolysis in any system with some water present.

18455

32

In essence, disappears relatively rapidly after application,
probably within two months in a forest environment. The evidence also
seems firm that deposited on the ground will not move into a water
course, and will net'leach into deeper layers.

 

It is difficult to separate physical behavior after application from
effects on lower organisms, because there is a very close interface be-
tween an agent bound to soil or in water, and insects or fish. Lower
organisms are of direct importance in assessing environmental impact
of any introduced chemica1:_ Eood chain effects may predict repercussions
at higher levels, and specifically sensitive organisms may be useful in-
dicators of pollution. Ecological displacement of any kind must be ac-
cepted only after careful examination, whether secondary to the intended
impact of the chemical on plant Species or as a result of direct toxic
effect of the applied chemical.

 moo

The various derivatives of differ in toxicity to fish. The
parent herbicide, the amine salts and isooctyl esters are relatively limited
in toxicity, but the propylene glycol butyl ether and butoxy ethanol esters
are quite toxic to some species. There is a large number of gross effect
Studies on various aquatic species; these generally use an end point of
death or immobilization. Shellfish studies measure shell growth as an
index of effect.

Standardization of methods has not received as much attention as
seems justified. Study periods vary from 24-96 hours, water temperatures
sometimes appear outside the normal acceptable range for the species, and
most evaluations seem to be carried on under static conditions. Nonethe-
less, the relative gross effects can be useful in reference to a given
application in the field. As a very rough approximation, a 2 kg/hectare
application over a water course would produce a concentration of about 2
mg/L or 2 ppm, if an assumption of dilution in the upper 10 cm of water
is accepted. Flowing water will diminish the concentration below detectable
limits in a few hundred yards (Evans and Duseja, 1973), and application
OVer deeper water will dilute accordingly.

Concentrations of or its derivatives found to have no ef-
fect are liSted below as acid equivalents:

 

No Effect
Exposure Acid Equivalent
Species Period Concentration Reference
killifish 48 50 Butler, 1963
mullet 48 50 Butler, 1963
sea lamprey 72 2 Applegate et
al., 1957
bluegill 12 46 Hiltibran, 1967
Na salt

I.



33

 

CD
0Q
03 trout 24 4.1 Applegate et
i ?4 butyl ester al., 1957
'5 e4 bluegill 24 hr 4.1 Applegate et
- c; al., 1957
CD lamprey 24 4.1 Applegate et
al., 1957
bluegill 8 7 Hiltibran, 1967
c) isooctyl green sunfish 8 7 Hiltibran, 1967
C) ester bluegill 12 0.7 Hiltibran, 1967

Median lethal concentrations of derivatives are similarly

 

tabulated:
Exposure Lcso
Period Acid Equivalent
Species (hours) Concentration Reference
bluegill 48 144 Hughes and Davis (1963)
DNA salt
bluegill 24 54 Davis and Hughes (1963)
TBA salt
bluegill 43 2.9 Davis and Hughes (1963)
Oleic-l,3?
propylene
diamine salt
bluegill 48 1.7 Davis and Hughes (1963)
isopropyl
ester
bluegill 48 31 Hughes and Davis (1963)
isooctyl
ester
bluegill 48 1.4 Hughes and Davis 1963
butoxyethanol
ester
bluegill 48 17 Hughes and Davis, 1963
PGBE esters spot 24 0.21 Cope, 1965
harlequin 48 1.0 Alabaster, 1969

While the more complex esters of tend to greater toxicity in
water at pH 6.5, they hydrolyze to the parent acid in about a day (Teasley
and Williams, 1970, as quoted by Kenaga, 1974).

1645i

34

Effects of on other aquatic organisms have also been measured.
In flowing salt water 1 caused no mortality in brown shrimp

and 2 was without effect on oyster shell growth (Butler, 1963). The _m 
PGBE ester of at 0.14 (0.09 acid equivalent) caused a 50% on p.
decrease in oyster shell growth after 96 hour exposure (Butler, 1963). 

Studies of terresrrial non-mammals seem to have been limited to PC
honey bees. Honey bees are an important component of the crop cycle and FA

in many areas are systematically managed for pollination. Phenoxy herbi-
cides, including fed in 60% sucrose-water solution at 10 

did not affect hatching, but reduced brood development when fed at 100
(Morton and Moffett, 1972). When sprayed in a water carrier at field
application rates, was non-toxic. (Petroleum solvents alone
caused high mortality during the first day after spraying.)

Aerial spraying did not result in herbicide accumulation in honey
sacs or colonies (Moffett and Morton, 1972). The point was made that the
primary damage would be failure of flowers and loss of nectar, rather
than direct toxicity. When fed directly to newly emerged bees, the
phenoxy herbicides were essentially non-toxic at concentrations up to
1000 in the diet (Morton et al., 1972).

2.4-0

Toxicity to Humans

 

For some reason there have been reported several incidents of 
poisoning in humans, but virtually no reports of human intoxication by
Probably the best known was a suicide by a single dose of 
DMA salt of which more than 90 mg/kg was estimated to have been retained
by the victim, after extensive vomiting (Nielsen et a1., 1965). An
accidental poisoning with a formulation containing 49% 
thiocarbamate, 9% kerosene, and 36.5% isooctyl ester caused symp-
toms and laboratory findings attributable to 2,4-0 intoxication. Among
these were twitching and paralysis of intercostal muscles, hemoglobinuria,
and myoglobinuria. No evidence of peripheral neurological damage was
evident during 36 months of follow-up observations (Berwick, 1970).
Goldstein et a1. (1959) reported on three cases in which esters
were absorbed through the skin, resulting in peripheral neurological
changes. One patient spilled 60 ml of a 10% solution of eSter on
the forearms and did not wash. Unusual fatigue develoPed in a few hours,
and over 10 days after exposure there was extended nausea and vomiting,
with considerable weight loss. A second exposure caused similar 
than pain and numbness in all digits, loss of skin from the palms, and
within six weeks there was substantial general neural damage. Another
patient eventually developed metacarpal pain and swelling in bath hands,
and later became partially paralyzed. The third experienced gaStrointes?
tinal disturbance and vertigo, then paresthesia in the arms and legs and
persistent general muscle fasciculations._ In two of the cases, there
were second exposures which appeared to cause greater impact than the
initial event. Similar cases have been described by Todd (1962), Berkeley
and Magee (1963), and Wilson (1956).

isles.

ll.

DOW 0011? 822

Seabury (1963) attempted use of in treatment of a terminal
case of disseminated coccidoidomycosis, on the rationale that the 2,4-0
as a plant hormone might alter the course of the fungus infec-
tion. At the time of the treatment (1949) there was no therapeutic agent
the disease. In 24 treatments over a period of more than a month the
dosage was raised to a final treatment of 3600 mg. No prior doses, up
to 2000 mg caused any response, but the final administration caused
extreme quiescence, and fibrillation of muscles in the face and hands,
followed by deep stupor and reflex failure. The patient recovered from
the within 48 hours and died of the fungus disease about two weeks
later.

General Toxicity to Laboratory Animals

 

One of the earliest published studies of toxicity was by
Eucher in 1948. She found an acute L050 of 280 mg/kg in mice, and found
that single doses of 150-200 mg/kg would produce a myotonia persisting
several hours. The animals remained awake and alert, but when moved ex-
hibited gross incoordination. They were capable of working out of the
with continued exercise, but if allowed to remain quiet the myo-
tonia would recur. The chemical also caused diarrhea in mice at the
higher doses, and gastrointestinal and upper respiratory irritation in
dogs. Mice were able to tolerate daily doses of 1/2 L050 for three
months. Hill and Carlisle (1947) established L050 values for several
Species: mice, 375 mg/kg; rats, 666 mg/kg; rabbits 800 mg/kg; and guinea
pigs, 1000 mg/kg. They also treated monkeys and were able to give up to
214 mg/kg without severe residual effect. Twice that dose caused vomiting,
incoordination, and loss of muscle tone. Rowe and Hymas (1954) summarized
lethality data on and five salts or esters; in general the lethal
doses were very high, with the exception of a few dogs given it-
self, for which the was estimated at 100 mg/kg. The latter data
was apparently derived from the observations of Drill and Hiratzka (1953).
Hansen et a1. (1971) conducted a two-year feeding study on rats, at dosages
of to 1250 in the diet, without appreciable change in growth,
hematologic values, or organ weight. In the same study dogs were main-
tained for two years with little effect at doses up to 500 2,4-0.
Drill and Hiratzka (1953) found that 20 mg/kg daily was fatal in 3 of
4 dogs within 49 days of a 90?day study.

Grazing species of animals might be expected to suffer exposures in
the field from eating treated vegetation. Palmer and Radeleff (1964)
evaluated a series of herbicides at high doses in limited numbers of
ruminants. One sheep tolerated 481 daily doses of 100 mg alkanola-
mine salt, another was unaffected by the same treatment with propy-
lene glycol butyl ethyl ester. The first compound was lethal after
7-500 mg/kg doses; the second was lethal after 9-250 mg doses. A bovine
tolerated 112 doses of 50 mg of the alkanolamine salt, but another de-
veloped digestive problems after 80 doses of 100 mg/kg. Palmer (1963)
administered daily-doses of 50-250 mg as the alkanolamine salt
to a group of steers. The steer given 100 mg/kg developed ruminal atOny
after 86 days, presumably as a result of the herbicide. Animals given
200 and 250 mg/kg became intoxicated in 34 and 15 days, respectively,
with dessicated mucous membranes and tendency to nosebleed when restrained.

36 15459

Ruminants on sprayed pasture or cows given 5.5 gm daily did net show
clinical evidence of toxicity, nor did production decline. 2,4-0 appeared
in serum of a cow fed 5.5 gm/day for 106 days, but none was passed in
milk (Mitchell et al., 1946).





Chickens and other fowl also appear relatively insensitive. Bjorn
and Northen (1948) found no impairment in weight gain at doses up to 28 mg
2,4-0 alkanolamine/kg three times weekly for four weeks; at 280 mg/kg
there was a marked decrease in gain. Single doses of 765 mg/kg were
lethal, 380 mg/kg was not. did not cause decrease in growth rate
at dose rates up to 1000 in the diet, but at 7500 growth essen-
tially stapped (Whitehead and Pettigew, 1972). The acute L050 was
estimated at 900 mg/kg. Whitehead (1973) tested dietary at concen-
trations up to 100 mg/kg diet, and found that at dietary levels of 10
mg/kg diet or greater, growth rate was depressed. Because food conver-
sion efficiency was not affected, it was suggested that palatability was
affected resulting in decreased consumption. Chickens will discriminate
against contaminated food (Whitehead and Pettigrew, 1972). The
U.S. Fish and Wildlife service tested a number of chemicals by feeding
to various game bird species in the early 19605. 2,4-0 acetamide given
to young quail at 2500 caused 72% mortality in 12 days, the butoxy?
ethanol ester at 5000 caused 28% mortality in 135 days, and 2500 
dimethylamine salt caused 12% death in 138 days. In older birds 2500
was almost without effect after 50 days, and after 111 days of 1000
feeding. Tests on Coturnix indicated a similar tolerance (Stickel,
1964). More recently, Hill et a1. (1975) have shown the LC50 for 
acetamide in the diet of bob white, coturnix, pheasant, and mallard to
be in excess of 5000 ppm. The butoxy ethanol ester and DNA salt have a
similar low toxicity.

838 "111500 nice

A number of other biological effects of have been found ex-
perimentally, usually at very high doses. Dybing and Kolberg (1967) sug-
gested that 2,4-0 was actively reabsorbed, and if so would competitively
decrease the clearance of p-aminohippurate without interfering with crea-
tinine clearance. Studies with rabbits given 100 mg priming doses, then 2
mg produced decreased PAH clearance at doses of 62-115 mg/kg.
Chang et al. (1974) studied a number of effects on rat liver following
treatment with 2-5 gm/kg 2,4-0 over 4-7 weeks. Glycogen content was 50-
100% higher, and liver nuclei RNA more actively i?_vitro than
did controls. DNA content per liver was decreased. In this work, 
was compared with the latter compound caused increased liver
weight and protein accumulation, while decreased liver weight and
lessened RNA content, with no protein accumulation. Speculation is jus-
tified that TCDD in was responsible for the difference. A dietary
supplement of 60 to lambs had no effect on rumen liquor or sheep
or serum pr0teins. Weight gain was decreased over a 12 week
feeding period (Abou-Akkada et al., 1975). The same authors (1973) had
previously found that 2,4-0 did not alter ruminal microbial funCtion.

At fairly high doses (80 mg/kg/day, for seven days) 2,4-0 increases 1311
intake by the thyroid. The effect only occurs in a normally functioning
thyroid gland; it was not seen after hyp0physectomy or iodine depletion
(Florsheim and Velcoff, 1962). The effect is apparently due to lowered
thyroxine binding to serum protein (Florsheim et al., 1963) but whether
this is secondary to 2,4-0 binding or is a specific pharmacologic effect

is not known. 
37 1643:50

4 

There has been limited in_vitro study of Weiss and Beckert
found stimulated mitotic activity and increased chromatin after cultured
monkey kidney cells, Girardi heart cells, and trout gonad cells were
exposed to 10 or 50 for 72 hours. inhibits mevalonate
incorporation into non-saponifiable lipids of liver, but only at concen-
trations in excess of 1 mM (Olson et a1., 1974). Oxidative phosphory-
lation in rat mitochondria appears to be sensitive to concentrations
as low as 10?4 at 10'3 respiration was normal but decreased to
20% of normal (Brody, 1952). In L929 cells in monolayer culture, 
causes triglyceride accumulation when present at a concentration of 500
ug/ml (Kolberg et a1., 1972) and inhibits all growth at 50 ug/ml (Kolberg
et al., 1971).

 

Dow}. {.0011 82

at high doses is used also as a chemical inducer of experi?
mental myOtonia, as a model of the congenital disease (Iyer et al., 1977;
Eyzaguirre et al., 1948). The action is apparently due to an increase in
membrane resistance and reduced chloride tranSport. Electrical activity
of the brain was found to be reversibly inhibited by doses of 200 mg
in rats (Desi et a1., 1962). The chemical also produces a
primary myopathy that is useful in study of the family of diseases which
includes white muscle disease in lambs (Heene, 1969). These effects are
not of toxicologic significance except in cases of massive acute intake
resulting in myoneural 

Effect of on ReproductiVe Function

 

As with the possibility that has teratogenic poten-
tial was first studied by the Bionetics Research Laboratories study
(Bionetics Research Laboratories, Inc., 1970). The data were suggestive
that incidence of failed lower jaw formation was somewhat greater than
that resulting from the DMSO carrier. Schwetz et a1. (1971) measured
teratogenic and fetotoxic effects of and two esters on rats. The
higher daily doses (75 mg 75 mg propylene glycol butyl ester
of 87.5 mg isooctyl ester of each just below the
maternal toxic dose) caused decreased fetal weight, subcutaneous edema,
delayed bone ossification and wavy ribs. Most of these changes are
fetctoxic rather than teratogenic. The last two are developmental ef-
fects but have no effect on survivability. No teratOgenic responses were
found at any dose.

There is some difference in definition of teratogenic response among
authors in the field. A variety of skeletal defects that do not interfere
with postnatal survival were found by Khera and McKinley (1972); most ef-
fects observed were wavy ribs or fused Sternum. The increased incidence
of these changes was evident at doses as low as 25 mg/kg/day. 
teratOgenesis was Studied by B?ge et a1. (1973) but only in presence of
The mixture caused some teratogenesis, but was less effective
than alone, so the impact of in the system was difficult
to evaluate.

Ji, Hamsters are subject to a teratOgenic effect of high doses of 
Collins and Williams (1971) found that from three different sources

isasi

38




caused a low incidence of anomalies, usually fused ribs, at doses of 100 

mg/kg/day through days 6-10 of gestation. There was no satisfactory CD

dose response relationship. Dietary 2,4-0 at 500 and 1000 did not FA

alter reproductive function in a three-generation, 6 litter study with Ff,
rats. Percentage of pups surviving to weaning and weanling weight was 
decreased at 1500 ppm, howeVer (Hansen et al., 1971). bo,?
Cf!

Sheep are apparently not subject to 2,4?0 induced teratogenesis.
Binns and Johnson (1970) adminiStered 2 grams daily fer 50, 60, and 90
days following breeding and caused no malformation. There were presum?
ably 6 sheep per group but the number in this specific experiment was
not stated.

Eggs are peculiarly vulnerable to exposure to herbicides and several
studies have been directed toward defining the hazard of to chicken
or game bird eggs. An injected dose of 10 mg caused 50% mor-
tality, mg/egg resulted in 30% loss, and at 0.5 mg/egg 90% of the eggs
hatched. No defonnities occurred at any dose (Dunachie and Fletcher,
1967). A later study by Dunachie and Fletcher (1970) confirmed these
findings for and (2,4-dichlorophenoxybutyric acid).

In contrast, Lutz-Ostertag and Lutz (1970) sprayed pheasant and
grouse eggs with at rates commonly used in the field and caused
embryonic mortality and terata. Somers et al. (1973, 1974) were not able
to support these findings after spraying mixtures of and picloram at
usual field rates (2.8 kg/ha) and and on hen eggs (1973)
and pheasant eggs (1974) at 10 times field concentrations (11.2 kg/ha).
They found no "adverse effect on hatching success, incidence of malformed
embryo or subsequent chick mortality". Kopischke (1972) also found no
effect on pheasant eggs sprayed at field concentrations, but found that
diesel fuel as a carrier blocked hatching completely. Dipping hen eggs
in 1% for 10 seconds, then continuing incubation was similarly
ineffective (Gyrd?Hansen and Dalgaard-Mikkelsen, 1974).

An interesting observation of distribution in mouse fetuses
has been made by Lindquist and Ullberg (1971). Labeled given late
in gestation accumulated early in the yolk sac, passed on to the fetus
and was almost completely eliminated by 24 hours after administration.
Distribution among tissues was non?selective, and concentrations tended
to parallel those of the dam, perhaps explaining in part the lack of
teratogenic effect.

Carcinogenic and Motagenic Potential of 

Innes et al. (1969) screened 120 compounds for tumorigenic properties
in mice. 2,4-0 and several of its esters were included; none caused in-
creased tumor incidence. Hansen et al. (1971) carried out a carcino-
genesis study in rats and concluded that although tumors were found,
the observed incidence did not support a finding that 2,4-0 is carcino-
genic. There seemed to be some inconsiStencies in interpretation that
any have confused the issue. Apparently no other evaluations of cancer
potential of have been made. A number of mutagenic screens have
included 2.4-0, however. Jenssen and Renberg (1976) found that 2,4?0

16462

39

 

would not induce increased micronuclei in mouse bone marrow 

but the compound did depress mitotic activity. Sex?linked lethality
assay of in malevDrosophila was also negative for mutagenic activity
(Vogel and Chandler, 1974). Styles (1973) treated rats with 2,4?0, then

used serum from the animals in'a host mediated assay with histidine-requiring
S. Typhimurium mutants. No effect of was evident. In a screen of

110 compounds with the "Ames test,? using eight histidine?requiring mutant
strains, Anderson et a1. (1972) was unable to detect mutagenic activity

by 2,4-0.

Absorption, Metabolism, Tissue Distribution, and Excretion of 

was absorbed rapidly from the lung of rats in which the herbi?
cide was injected intratracheally as 0.1 m1 of 0.01?10 mM solution. The
animals were sacrificed from one half to 120 minutes later. The half-
time for 2,4?0 absorption was 1.4 minutes, and concentration had little
influence on the rate (Burton et a1., 1974). When ingested, 2,4-0 is
absorbed primarily by the portal circulation; the drainage
accounts for very little 2,4-0 absorption, as might be expected from the
limited fat solubility of the herbicide (Sieber, 1976). Blood concen?
tration of 14C from radiolabeled 2,4-0 administered orally to sheep has
been shown to rise very rapidly (Clark et a1., 1964). The rate of ab-
sorption of from the rumen is not known; the immediate elevation
in blood 14C suggests that at least some material was removed directly
from the rumen. However, Gutenmann et a1. (1963) fed 5 2,4-0 to
cattle and found that the concentration in the rumen contents decreased
from 3.5 to 0.5 over a 24 hour period, which does not suggest rapid
absorption, but rather dilution with passage of rumen content. The
chemical did not disappear in an artificial rumen.

Kohli et a1. (1974) gave 5 mg orally to six human volunteers and
found peak plasma concentrations by seven hours; the average plasma
concentration one week later was about 10% of the peak concentration.
Half time for excretion was calculated at 33 hours.

Clinical observations of human patients who had extensive skin
contact with 2,4?0 showed systemic toxic responses within a few hours
[Goldstein et a1., 1959; Todd, 1962), indicating ready absorption
through the skin.

'Application of as the DMA salt, isooctyl ester and butyl
ester to the body surface of rabbits was relatively ineffective (Kay et
al., 1965). Treatment was with 15 ml of aqueous or oil solution applied
on a 4 3 gauze patch, covared with plastic film. Contacr was
seven hours daily, five days a week for three weeks; concentrations of
2.4-0 were 0.626 and 3.13% acid equivalent. Some animals were treated
on abraded skin. A few animals died during the experiment but the
pathology and were not typical of poisoning; most of the
fatalities were among the animals with skin damage. No neurological
lesions were found, nor were any other observed parameters changed.

The only lesions observed were at the site of application.

355483

40

There seems to be general agreement, other than the report by Kohli
et a1. (1974), that significant muounts of do not remain in an animal
for much more than one or two days. Zeilinski and Fishbein (1967) com-
pared whole body residence times of two esters of and the 2,4-0
acid after subcutaneous injection of 100 mg/kg to mice. Of the butyl
ester, 95% was gone after 6 hours, and in animals that had been pre?
treated with five similar daily doses, the rate of disappearance was fur-
ther enhanced. The isooctyl ester was considerably slower to disappear,
with a half time of somewhat less than four hours, and the acid itself
was retained longer. The authors did not Specifically note
whether hydrolysis of eSters to acid was identified as part of the
metabolic process, but the method appeared to account for the disappear-
ance of all forms of the compound. As a comparison, the disappearance
half-time of acid was on the order of 20 hours.



 

Khanna and Fang (1966) adminiStered to male and female rats
at very high (80 mg/rat) and relatively low (1 mg/rat) doses and found
tissue residues to be greatesr at about 6 hours, and almost undetectable
by 30 hours after treatment. Extremely high concentrations were found
in the stomach but there was no information about separation of stomach
content from the tissue proPer. No label was fOund in respiratory gases.
A very small fraction of urinary label was found to be an unidentified
metabolite, but the data indicate that almost all is excreted with?
out change by rats.

Although binds reversibly to bovine serum albumin (Kolberg et
al., 1973) and therefore probably to other plasma proteins, there seem
to be no specific tissue binding sites as are found in plants. Morre
(1974) has examined fish muscle and rat liver after exposure to 2,4-0,
seeking specific binding sites and has concluded that none exist.

apparently does not move into milk of lactating animals in
significant amounts, whether treated by direct administration of the
herbicide or by grazing on treated land (Bjerke et al., 1972; Gutenmann
et al., 1963; Bache et al., 1964; St. John et al., 1964; Klingman et al.,
1966). Under forcing conditions, however, (1000 in diet), the
level of residue in cows milk was forced up to 0.06 ppm, and were
still barely detectable seven days later.

A substantial capacity for conjugation of has been des-
cribed elsewhere in this report (Nony et al., 1976). Grunow and B?hme
(1974) have found that the ability to conjugate 2,4-0 with glycine and
taurine exists but is considerably less than that for In dog-
fish and flounder, however, a major fraction of urinary 2,4-0 is-excreted
as the taurine conjugate (James and Bend, 1976), and in three of the four
dogfish studied 10-20% of the label was found in bile after 48 hours.

In spite of the rapid loss of by fish, persistence of the
herbicide or its products may be prolonged. Treatment of pond weeds with
up to 9 kg/ha usually left no detectable residues in fish by 28 days
post-application, even at the highest rates (Schultz and Harman, 1974).
Schultz (1973) has found, however, that some unidentified products may
be present in fish 2-3 months after treatment..

16464

41

 

The rapid appearance of intact 2,4?0 in urine is apparently mediated
by a presumably active and saturable tubular excretion mechanism (Erne
and Sperber, 1974). The extensive excretion of in urine has been
noted in steers (Lisk et al., 1963), sheep (Clark et al., 1964), rats
(Sieber, 1976), and humans (Kohli et al., 1974), and apparently is rapid
enough to remove all but massive exposures before significant damage
can occur.

Behavior of in the Environment and EffeCts on Submammalian Species

This report is primarily concerned with effects of herbicides on
non-plant species, but a cursory qualitative survey of literature on
field persistence is useful in judging how long a given amount of herbi-
cide may remain available.

Water concentrations are a major concern, in relation both to aqua-
tic species and to water supplies for the human population. 2,4-0 ap-
plied in a watershed area appears in streams to a very limited extent
(White et al., 1976; Krammes and Willetts, 1964). Movement into sub-
snrface water flows also appears negligible (White et al., 1976). In
aerated lake water persists up to 120 days, but in lake mud 
hydrolyzes very rapidly, because of microbial activity (Aly and Faust,
1964). Half-time of disappearance seems overall to be much less
than two weeks in aqueous systems.

For some applications on water, relatively massive applications of
up to 40 lbs. per acre are necessary. Two reservoirs on the Tennessee
River were treated at 20 and 40 lbs/acre depending on the nature of the
water milfoil infeStation. The treatments did not affeCt fish or other
fauna, and had little adverse effect on most other plants. Plankton did
retain significant.amounts of herbicide over extended periods, and 2,4-0
was detectable on occasion in finished domestic water from the reser-
voirs (Wojtalik et al., 1971).

In soils, 2,4-0 has been found to have a half-time of 4-5 days by
Altom and Stritzke (1973). White et a1. (1976) found that 95soil would disappear in 7 days. However, an in_
vitro study by Alexander and Aleem (1961) indicated that would be
detectable for as much as 94 days in one type of soil and only 23 days
in another. The preparation included 100 m1 of nutrient medium and 4
of soil as an inoculum; whether such a system represents actual break-
doWn conditions in soil is possibly questionable. Sufficient material
to cause detectable phytotoxicity persists for about a month, according
to Mullison (1972). In areas where is used annually, the break-
down may be more rapid in the later years than following the first
treatment (Hurle and Rademacher, 1970). Wiersma et a1. (1972) reported
a total of 28 which had been treated with were found to have from
0.01 to 0.03 2,4-0 present. It seems unlikely that would per-
sist in either water or soil for more than a month.

has been used extensively in control of aquatic weeds, par-
ticularly water hyacinth and water milfoil, and has probably been sub-
jected to more study of aquatic toxicity than any other herbicide. A

-ieass

42



number of short term screens have been conducted with fish. AlabaSter 
(1969) reported on 164 compounds, including salt, which I
was found to have a 24 hour LCSO of 1160 for harlequin fish, and the 
butoxyethylester of which was much more toxic with a 24 hour L050
of 1 ppm, for the same species.

.
. .?nu

6385-11300 moo

The 24-96 hr LCSO of PGBE esters of for rainbow trout were
above 1 (Cope, 1964). For mullet and killifish the LC50
(24 hr) was 5 for borh the PGBE and butoxyethanol esters. The parent
acid was ineffective at 50 (Butler, 1963).

Oyster shell growth was 50% inhibited by 3.75 butoxyethanol eSter,
but the acid and dimethylamine salt were not effective at 2 (Butler,
1963). Exposure to the DMA salt for 24 at 2 caused no mor-
tality in shrimp; 48 hour exposure caused 10% lethality. The butoxy-
ethanol and PGBE esters caused no mortality to shrimp at 1 48 hour.
The ethyl hexyl ester of caused 38% decrease in oyster shell growth
at 5 over 96 (Butler, 1965). The considerable differences in
2,4-0 effect among various species were illustrated by Sanders (1970a)
acid has a 43 hour median response concentration (TLso) of 100
mg/L (100 ppm) for Daphnia and 3.2 mg/L for scud. (Crosby and Tucker
(1966) found the same value for Daphnia.) The PGBE eSter TL50 varies
from 0.1 for Daphnia to 2.6 for scud to more than 100 for crayfish.
The DMA salt TLSO is more than 100 in bluegill but TLso for the
butyl ether ester in bluegill is 1.1 ppm, and 100 for crayfish.

Stonefly naiads are less sensitive to itself (LCSO 96 hr 
15 ppm) than the butoxy ethanol eSter (LCSO 96 hr 1.6 ppm) (Sanders
and Cope, 1968). Sanders (1970b) has also reported a 96 hr TLso for 
amine of 100 for tadpoles.

The lethal effect on tadpoles is apparently limited, but Buslovich
and Borushko (1976) found that 2,4-0 sodium salt would inhibit metamor-
phosis of Rana temporaria tadpoles, and blocked thyroidin stimulation of
the process. The DNA salt was effective at 2 ppm. The authors specu-
late that antagonizes thyroid hormone.

Few studies of truly chronic exposure of fish to have been
made. Mount and Stephan (1967) compared the 96 hour LCSO for 2,4-0 butoxy-
ethanol ester (5.6 ppm) with various 10 month exposures, and found that
1/19th of the LCSO, or 0.3 could be tolerated by fathead minnows
without effect on growth or reproduction. Eggs were much more sensitive
than adults over 48 hour exposures. Schultz (1973) placed bluegill,
channel catfish, and largemouth bass in solution of DMA salt, labeled
with 14C at concentratiOns of 0.5, 1.0, or 2.0 mg/L Fish and
water samples were removed at intervals up to 84 days for analysis of 14C
and content.

Considerable radioactivity remained in fish tissues but apparently
none was associated with 2,4?0, suggesting extensive metabolism. The
first tissue in which radioactivity appeared was the gall bladder of cat~
fish and bluegills, and eventually 14C appeared in every tissue analyzed.

18465

43

nun-p.? .



 

In both catfish and bluegill, the amount of radioactivity in muscle
tended to increase through the collection period. No toxicity was evi?
dent in any of the exposed fish.

To relate concentrations of one to field condition, an applica-
tion of 2 kg/hectare (10000 M2) to water will give a concentration of 2
mg/L or 2 if the water is 10 cm deep. Deeper water will result in
further dilution. If applied to flowing water, water movement will
dilute the herbicide quite rapidly, and that material distributed on
soil and vegetation will tend to remain in place. Obviously a spill
or other accident presents different problems.

Among insects, the effects of on honey bees has receiVed the
most attention. Palmer-Jones (1964) has listed a number of investiga-
tors who concluded that is safe for bees, as well as others who
have shown toxic effects, some of which may depend on the species of
plant on which the herbicide is deposited. In a field invesrigation of
a three lb/acre application Palmer-Jones found that a 22% mortality oc-
curred in 48 hours after dusting. However, when bees at the hive were
heavily dusted directly, there was no mortality, leading to the conclusion
that the bees in the field were acquiring intoxicant by some mechanism
other than surface contact. Moffett et a1. (1972) sprayed caged bees
directly with various phenoxy herbicides at a one pound/acre rate with
very little effect. When included in the diet at concentrations up to
100 ppm, 2,4-0 did not decrease lifetime of bees and the ester used was
ineffective at 1000 (Morton et al., 1972). The herbicide does cause
decreased brood development when fed at 100 but has no reproductive
effect at 10 (Morton and Moffett, 1972).

Treatment of Coccinellid beetle larvae with 2,4-0 at a rate equal
to 8 oz. acid equivalent/acre caused greatly increased mortality, re-
gardless of the age of the larvae through day 12 at time of spraying
(Adams, 1960).

SILVEX

There appears to be a much more limited accumulation of data on bio-
logical effects of silvex and related compounds than is available for
other phenoxy herbicides. The descriptions of biological effects of
silvex will be categorized in two broader sections rather than the more
detailed entries prepared for the other agents.

Biological EffeCts of Silvex and Its Derivatives

 

The acute median lethal dose of silvex and its esters is quite high,
and falls in a relatively narrow range among species and among the deri-
vatives. Rowe and Hymas (1954) summarized data which had been established?
at that time. Silvex and its mono?, di?, and tri-prOpylene glycol ether
esters were of almost identical L050 in guinea pig of 1200 and 1350 mg/kg.
The lethalities to rats were also similar at about 650 mg/kg. The rat
appears to be most sensitive, and chicks and mice require doses similar to
the guinea pig.

1846??

44

 Moo



Subchronic (90 day) Studies of rats fed 10-600 mg/kg/day of the PGBE
ester resulted in more than 50% lethality at the highest dose. The time
of death ranged from 15-85 days. Growth was depressed in rats fed 300
mg/kg. There is somewhat of a paradox in the findings, because pathology
in the dead animals indicated malnutrition rather than herbicide toxicity.
A paired feeding study at 300 and 600 mg/kg/day indicates that depressed
growth was not entirely due to inadequate food consumption. Dose rates of
30 and 100 mg/kg/day caused an increase in liver weight but 10 mg/kg/day
caused no change. (Mullison, 1966). The study was followed by a 90 day
feeding of up to 10000 silvex (sodium salt) in the diet. 10000 is
1% of the diet, and if the animals consumed 10 gm daily and weigh 300 gm,
the dose rate would be on the order of 300 mg/kg/day. That dose rate was
highly lethal and was abandoned, but did produce swelling and granular
degeneration of hepatocytes and ultimate cellular necrosis. Renal tubule
cells were swelled and vacuolated, and seminiferous tubules were degenera-
ted. At levels above 10 (about 3 mg/kg/day) some growth depression
occurred. Mullison (1966) also fed Kurosol (potassium salt of silvex,
containing 53.3% silvex acid) for two years at concentrations up to 300
ppm. The highest dose caused a increased kidney/body weight
ratio in males, but 100 and lower dose rates caused no change in food
consumption, growth, gross and microscopic morphology hematology or bone
marrow. 100 was stated by the author to be equivalent to 2.6 mg/
kg/day of silvex.





Kurosol was without effect on beagles at a feeding rate of 56 
(about 19 mg/kg/day) over 2 years. Females fed 190 suffered some
hepatic degeneration and necrosis after a year of feeding, but at the end
of two years no damage could be found. At the 56 intake, there were
no changes over a very broad spectrum of hematologic, clinical chemistry
and morphologic analyses.

The liver damage at higher levels included hepatocyte necrosis, bile
duct proliferation, and bile pigment deposition throughOut the liver and
in the epithelium of kidney tubules.

In a subchronic study in cattle by Palmer et al. (1964), one yearling
Brahma-cross fed 100 mg silvex/kg/daily died after 29 days. Two other
animals given 25 and 50 mg/kg for 73 days showed no evidence of toxicity.
A 90 day experiment with 50 mg/kg/day, either by drench or by injection
into a rumen fistula resulted in death of one of the latter group of
three animals. Two of those treated orally deve10ped severe inflammations
in the parotid area, apparently due to local irritation.

The survey by Innes et al. (1961) screened silvex against two strains
of mice for 18 months. The concentration of silvex was 121 ppm, which was
the maximum tolerable dose rate, and there was no increase in tumors in
either strain. The mutagenic screen by Anderson et a1. (1972) did not
detect point mutations resulting from silvex treatment in either T4 phage
or S. typhimurium (histidine requiring).

Silvex is teratogenic at high doses.' Courtney (1975) found that
398 mg/kg/day, on days 12-15 of gestation, caused 3% cleft palate in the

164188

45

 

group given silvex in DMSO subcutaneously, and 7% where given orally in
corn oil. Fetal mortality increased to 25% in the group treated subcuta-
neously.

The Dow Chemical Co. has also conducted a series of teratology studies
with silvex. Dose rates of 75, 100 and 150 mg/kg/day from day 6 to day 15
caused several cardiovascular anomalies; 50 mg/kg/day caused retarded
ossification in the sternum and skull (Dow Chemical Co., 1972). The no
adverse effect level was considered to be 25 mg/kg/day. The PGBE ester
caused skeletal changes at an intake of 50 mg/kg/daily but no changes
were found after 35 mg/kg/day (23 mg/kg silvex acid equivalent).

Birds seem peculiarly resistant to the herbicides. For example,
DeWitt et al. (1963) fed 5000 BEE ester of silvex to young bob white
quail, pheasant and mallard ducks. The average lethal intakes were 9350,
9240 and 21,000 mg/kg, and time of death varied from 10 to 100 days.
lbllards were able to consume 100 for 100 days with no lethality, but
reproduction was impaired.

Silvex acid at 5000 in the diet was tolerated for 5 days with
no mortality by Coturnix. The 5 day L050 for pheasants was 4500 of
silvex BEE ester, for bobwhite quail the L050 was 30 over 5 days,
for Ceturnix it was in excess of 5000 ppm, for pheasants LDSO was less
than 3000, and no mallards died after 5 days on 5000 ppm. All birds were
two weeks old at the initiation of the study (Heath et al., 1972).

Stickel (1964) reported US Fish and Wildlife studies in which bob?
vhite were fed 1000 of an unsPecified silvex ester; 50% were dead by
day 34 with average total dose of 17000 mg/kg. Of a group fed 5000 ppm,
the average daily dose was 2300 mg/kg. Half of the birds survived more
than 4 days, 40% survived more than 10 days, and an additional 4% died
during the remaining 14 days. The average dose to birds that survived'24
days was 54,500 mg/kg.

These findings indicate that wild fowl should not be adversely af-
fected by any field application of silvex, or for that matter even a gross
over-application.

Effect of Silvex on Aquatic and Invertebrate Species

 

SiIVex has become an herbicide of choice for control of surface and
underwater water needs. As a consequence, there has perhaps been more
attention paid to its impact on aquatic Species than any of the other
phenoxy herbicides. There is an unfortunate diversity of experimental
methods and conventions for expressing concentration and other factors
which sometimes makes comparison of data difficult.

Table 1 has been constructed to simplify consideration of the various
reports of toxicity to aquatic species.

Among factors that influence toxicity to fish are the chemical form

of the agent, whether the formulation is granular or liquid, and water
hardness. Generally the e5ters are more toxic, and liquid formulations

46

it

pu

Table 1. Gross Toxicity of Silvex and Its Derivatives to Fish

 

 

 

Concentration
(Acid Equiv.)
SDecies Form of Silvex Effect Period Reference
Bluegill BEE ester 0.36 LC50 48 Cope, 1964
BEE ester 1.7 LC50 48 Hughes and Davis, 1966
Isooctyl ester 1.4 . LC50 48 
(young) salt liquid 83 LC50 48 Hughes and Davis, 1965
salt granular 100 48 Hughes and Davis, 1965
(fry) PGBE ester 0.3 . tolerated 96 Jones, 1962
salt, liquid 100 tolerated 96 Jones, 1962
salt, granular -150 tolerated 96 Jones, 1962
(eggs) PGBE ester 10 no effect Wilber and Whitney, 1973
on hatch
(fry) PGBE ester 5 100% lethal 36 Wilber and Whitney, 1973
PGBE ester 1 no effect 80 days Cope, 1964
PGBE ester 3 liver degen. 2 weeks Cope, 1964
salt 10 no effect 2 1/2 mo Cope, 1964
salt 75 35% lethal 2 1/2 mo Cepe, 1964
Stoneroller salt 25 95% hatch 72 Hiltibran, 1967
(eggs) PGBE ester 5 45% hatch 72 h_ Hiltibran, 1967
Chorus frog BEE 20 LC50 24 Sanders, 1970b
(tadpole) BEE 10 LC50 96 Sanders, 1970b
Fowler's toad BEE 22 LC50 24 Sanders, 1970b

(tadpole)



ease-11004400

 

  
 

.Hdbiibasg

7- .



I

are more toxic. Data on water hardness is contradictory; but the ten?
dency to greater toxicity in soft water seems to be associated with the
salt rather than the esters (Surber and Pickering, 1962).

Distribution rates for silvex treatment of aquatic weeds may be as
high as 40 lbs/acre. An assumption of dilution thr0ughout an 8 foot depth
is'apparently conventional, which would give a concentration of 1.8-1.9
ppm. If this concentration of an ester formulation were to hold in a
given area, it would clearly be lethal to many fish. The salt should have
up to a 100 fold safety factor for fish, depending on mineral content of
the water. Such application is not a consideration in forest operation,
but accidental overwater dilution could occur. Terrestrial application is
at rates at least 10 fold less than 40 lb/acre but many water courses are
much shallower than 8 feet, thereby leaving a similar hazard. If flowing,
dilution will be rapid and periods of exposure should be limited. In
static water the agent will diffuse away from the application site with
time except in small shallow ponds, where some damage might possibly occur.

Lower aquatic organisms have also been well studied because of
silvex use on water weeds. Representative though not comprehensive data
is compiled in Table 2. Again, it appears that the salts of silvex are
much less toxic than the more complex esters. The most vivid contrast
is in the effect on Daphnia magna, which is 50% immobilized by 100 
silver potassium salt, and by only 0.18 silvex PGBE ester. Some
species, however, are particularly resistant to even the PGBE ester of
silvex. The crayfish LCso is in excess of 100 ppm; for a number of other
organisms this factor is less than 1 ppm.

It is not necessary to catalogue the entire literature on silvex
effects on either fish or other aquatic species. The range of effective
concentrations is well illustrated as well as the extent, if not details,
of Species differences. 

Effects of silvex on terrestrial insecc life have been evaluated
in honey bees by USDA workers at Tucson. All of the phenoxy herbicides
including silvex were found to be relatively non-toxic to bees when
applied in water at field concentration (Morton et al., 1972; Moffett
et al., 1972). When fed at 100 or 1000 ppm, silvex reduced brood pro-
duction, but at 10 ppm, no adverse effect was found. In each case, when
the toxicant was removed, the colonies regained their original reproduc-
tive efficiency (Morton and Moffett, 1972).

bbtabolic Fate of Silvex in Mammals

There have been few studies of the disposition of silvex after ab-
sorption by mammals. Bjerke et a1. (1972) fed silvex at 1000 in feed
and found 0.12 2,4,5-trichlorophenol in milk and 0.16 
in cream. In one week off c0ntaminated feed the concentrations decreased
below 0.05 ppm. Leng (1972) in a review, reported work in which 
was not detected; however, residues of siIVex were substantial in the
liver. After feeding 300 silvex 28 days, 4 were found in liver,
18 in kidney, 0.6 in muscle and 0.9 in fat. After 2000 in the
diet, 12, 30, 2 and 4 were found in the respective tissues. In a

154%7l

48

6t

z?agl

Table 2. Gross Toxicity of Silvex and Its Derivatives to Invertebrate Aquatic Species*

 

 

Concentration
Form of (Acid Equiv.)
SEecies Silvex gem Effect Period Reference
Daphnia magna PGBE ester 0.18 LC50 48 Sanders, 1970a
BEE ester 2.1 LC50 48 Sanders, 1970a
salt 100 IC50 26 Crosby and Tucker, 1966
Scud PGBE ester 1.8 LC50 24 Sanders, 1970a
PGBE ester 0.8 LCSO 96 Sanders, 1970a
. BEE ester 1.2 LC50 24 Sanders, 19703
Seed Shrimp PGBE ester 0.2 LCSO 48 Sanders, 1970a
BEE ester 4.9 LCSO 48 Sanders, 1970a
Glass Shrimp PGBE ester 3.2 LC50 48 Sanders, 19703
BEE ester 8.0 LC50 48 Sanders, 1970a
Sowbug PGBE ester 0.5 LC50 48 Sanders, 1970a
BEE ester 40 LCSO 48 Sanders, 1970a
Crayfish adult PGBE ester 100 LCSO 48 Sanders, 1970a
Daphnia pulex PGBE ester 210 1C50 48 Sanders and Cope, 1966
Simocephalus PGBE ester 2.4 ICSO 48 Sanders and Cope, 1966
Serrulatus 
Stonefly naiad acid 5.2 ICso 24 Sanders and Cope, 1968
- .34 1C50 96 Sanders and Cope, 1968
PGBE ester 5.6 IC50 24 Cope, 1965
.34 LCSO 96 Cope, 1965
Brown shrimp PGBE ester 0.28 LCSO 24 Butler, 1965
adult
[?50 median lethal concentration
1C50 median concentration to cause immobilization

*all first in star unless otherwise noted

   

  

Basianoctaoa

 

100113336

similar experiment Clark et a1. (1975) found roughly similar values, with
traces of in each tissue. In a single cow experiment, 5 
kuron was fed for four days at 5 (St. John et al., 1964). The kuron
hydrolyzed to silvex and was excreted as a salt in the urine. Urinary
concentrations peaked at 4.4 on day 4 and declined to 0.32 on day 6.
About 67% of the total fed was found in urine; feces were not analyzed.
No detectable residues of Kuron or silvex were found in milk and no kuron
as such entered the urine.

The pharmaco-kinetics of silvex have been Studied in rats given 5 or
SO mg/silvex/kg. The compound was ring-labeled with 14C. The higher dose
appeared to be saturating because plasma clearance was not linear, as was
the case at 5 mg/kg. Half-time for plasma clearance was 9-10 hours at
both doses. At the lower dose 76% of the labeled material emerged in bile
in 72 hours and 90% of the high dose. Almost all of the biliary excretion
was in the first 24 hours. Most of the material was subject to entero-
hepatic recirculation and eventually left the body in urine. Total siIVex
excreted in 192 hours in urine was 78%, and 16% appeared in feces (Sauerhoff

et al., 1977a).

Sauerhoff et al. (1977b) have also examined silvex diSposition by
humans. One mg/kg was given orally to seven men and one woman, and plasma
urine and feces were analyzed for 168 hours. Clearance from plasma and
appearance in urine were bi-phasic, each stage following apparent first?
order kinetics, maximum plasma concentrations occurred in 2-4 hours, 65%
of the dose was excreted in urine in 24 hours. Half times for the two
phases in plasma were 4 and 16.5 hours and in urine were 5 and 26 hours.
The largest amount recovered in feces was 

'50

at}:



U.)

REFERENCES

2 I



Abou Akkada, A.R., M.A. Hassan, and M.A. Naga (1973). Effect of some
pesticides and herbicides on the rumen micro-organisms of sheep.
J. Agr. Sci. 81:231-236.

?151500 Moo





.
- . '3 hi] .
 

Abou Akkada, A.R., M.A. Hassan, and M. Naga (1975). A note on the
effect of herbicides on the metabolism and performance of lambs.
Anim. Prod. 20:311-313.



2.89

Adams, J.B. (1960). Effects of spraying amine on coccinellid
larvae. Can. Jour. 2001. 38:235-283.

Alabaster, J.S. (1969). Survival of fish in 164 herbicides, insecticides,
fungicides, wetting agents, and miscellaneous substances. Internat.
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Adexander, M., and M.I.H. Aleem (1961). Effect of chemical structure
on microbial decomposition of aromatic herbicides. J. Agr. Food
Chem. 9:44-47.

Allen, J.R., J.P. Van Miller, and D.H. Norback (1975). Tissue distribu?
tion, excretion and biological effects of [14C]tetrachlorodibenzo-
p?dioxin in rats. Pd. Cosmet. Toxicol. 13:501-505.

Allen, J.R., D.A. Barsotti, and J.P. Van Miller (1977a). Reproductive
dysfunction in non-human primates exposed to dioxins. Presented
at 16th Annual Meeting, Soc. Toxicol., Toronto, 1977. Abstract 111.

Allen, J.R., D.A. Barsott, J.P. Van Miller, L.J. Abrahamson, and J.J.
Lalich (1977b). Morphological changes in monkeys consuming a diet
containing low levels of Food
and Cosmetics Toxicol. (in press).

Adtom, J.D., and J.F. Stritzke (1973). Degradation of dicamba, picloram,
and four phenoxy herbicides in soils. Weed Science 21:556-560.

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Environ. Health Persp. 5:221-225.

wright, F.C., J.C. Riner, J.S. Palmer, and J.C. Schlinke (1970). Metabolic
and residue studies with 2-dichloro-
propronate (Erbon) in sheep. J. Agr. Food Chem. 18:845-847.

Yang, K.H., P.D. Guiney, J.L. Seymour, and R.E. Peterson (1977). 2,3,7,8?
Tetrachlorodibenzo?p?dioxin (TCDD)-induced depression in biliary ex-
cretion of biphenyls (PCBs) in rats. Presented at
16th Annual Meeting. Soc. of Toxicol., Toronto 1977. Absrract 103.

70

Yoder, J., M. Watson, and W. Benson (1973). chromosome analy-

sis of agricultural workers during extensive occupational exposure
to pesticides. Mut. Res. 21:335-340.

Young, A.L. (1974). Ecological studies on a herbicide-equipment test

area. Technical Report USAF Systems Command, Eglin
AFB, Florida.

([4593 AA (NKT

Zielinski, W.L., Jr., and L. Fishbein (1967). Gas chromatographic measure-
ment of disappearance rates of and acids and esters
in mice. J. Agr. Food Chem. 15:841-844.

Zinkl, 5.6., J.G. Vbs, J.A. Moore, and B.N. Gupta (1973). Hematologic and
clinical chemistry effects of in
laboratory animals. Environ. Health Persp. 5:111-118.

16494

71

 

 

18.338
DOW 
?a 2 '0 .L .L


NHCHIGAN QGGJO

June 3, 1974

George E. 
1062? El Capitan Circle
Sun City, Arizona 85351

cc: J._Davidson, 9003

Traynor, 2030
R. Wessel, Washington,D.C.
E. H. Blair, 2020

Crummett, 574

Gehring, 1803

TOXICOLOGY OF POSSIBLE CONTAMINANTS IN 

On May 6, Lefty asked that the toxicity of possible oon-
taminants of be made available should EPA or EDF
people ask for such information. I have searched our
files for our information with the following results.
Note that the possible contaminant list was supplied by
Andy Watson. .

I found no tox- data on the following:

Amounts Likely
in wt. percent

0.4 acid
<0.02 2,3,6-trichlor0phenoxyacetic acid

3.2 acid
4.1 acid

acid

<0.05 2,3,6-trichlorophenol
2,5-dichloroquinone monomethyl ether
4,6-dichlororesorcinal monomethyl ether

0.03 4,5-dichlorocatechol monomothyl ether

0.02 2,6?dichlorophenocyacetic acid

0.26 2,S-dichlorophenoxyacetic acid

0.03 Z,4,6?trichlorophenoxyacetic acid

25:: 
3; I "z 
. 
AN UNIT OF THE DOW CHEMBCAI. COMPANY 
. 
15455

rm\



I 



-2-

The following contaminants had some data:
0.07 hichloronhenol



. 

Acute oral guinea pig in range of 0.3 to 2.6 g/kg
Acnegenlc actirity - none

2,4-dichlorgphenol

-

 



Acute=eral guinea pig in range of 0.3 t0 2.0 g/kg
Skin - old sample - caused skin burn in 15 sec.?
- new sample - non-corrosive by DOT test

Acnegenic activity - negative
Odor Threshold

Dow 0.02 
Taste Threshold

Dow Fish 0.005 

Lit. - water - 0.002 mg/l
Fish Tox.

LC100 to 100
Lit. values

Acute oral

in water

L050 0+ 4.5 g/kg
o? 3.67 g/kg

Rat

Mouse LDSO 1.6 g/kg
Chronic oral
6 mo. - mice
No effect 100 mg/kg/day

200 mg/kg/day
slight depression
slight depression

Adverse effect
Liver wt. -
SGPT 

2,5-dichlorophenol

 

 

Acute oral guinea pig in range of 0.1 to 2.8 g/kg
2,6-dichlorophenol
Acute oral guinea pig in range of 0.8 to 2.0 g/kg
Acute oral rat >2.0 g/kg
moderate superficial burn which should heal
in a week or so.
Skin - a burn Upon 24 hr. contact

- a superficial burn in 1% hr.
24-48 hrs. .

Ref. says 2,6?dichlorophenol may be a sex
hormone in a tick Species.

heals in

Lit.

(15493-



 

 

-3-

<0.02 2-Chlorophenoxxagptic acid

Acute oral - Rat in range of 1 to 3 g/kg

Skin - minor irritation

Skin absorption - no indication in skin tests
<0.09 4-Chlorophenoxyacetic acid

Acute oral - lhat 0; L050 1.54 g/kg

Acute oral - Chicks LDSO 1.60 g/kg

Skin - dry powder - no irritation
0.06 Z,4-0ichlorophenoxyacetic acid -

 

,Acute oral LDSO

Rat 375 mg/kg
douse 368 mg/kg
G. Pig 469 mg/kg
Dog 100 mg/kg Lit. Ref.

Chicks 541 mg/kg
Chronic oral

Rats fed 114 day in diet
0.03% - no effect level
0.1% - adverse effects - minor liver and
kidney path. and growth depression

Skin - Rabbit

Powder (100%) dry - no irritation

- slight irritation
Skin - Human

1 application for 4 to 5 days, then rest 3 wks.
followed by challenge 24 hr. CXposure

?not a sensitizer?
no to slight irritation depending on
sample used

Chloracne - negative

17842.9?





<0.02







.. 4-

Teratology and Fetal Toxicity
87.5 mg/kg/day to dam during 6-15 day
fetal toxicity positive
teratology_? negative
Odor Threshold
60 

Metabolism

72 hrs. - Sheep 96% excreted - urine unchanged
1.4% excreted - feces unchanged

3,4?Dichlorophenoxyacetic acid

Acute oral - Rat 250-500 mg/kg

- a transient burn lasting 2-3 days, completely
healed in days.

Skin - 100% dry - no irritation
- 10% sol. - slight irritation

Skin absorption no indication in skin tests run.

2,4,S-Trichlor0phenol

Acute oral - Rat male - LDSO-2830 mg/kg body wt.
Lethality - Rat - female - LDSO-2460 mg/kg body wt.

Chronic oral Rat Purified 2,4,S?trichlorophenol
was fed in food at 100, 300, 1000, 3000 and 10,000
for 98 days. No adverse effects were detected
at levels of 1000 or less; 3000 and 10,000 
caused mild diuresis and slight pathological changes
in the liver and kidney. 

Irritation Rabbit - The product caused conjunc-
tival membrane burns, severe conjunctival chemosis,
slight to moderate chemosis, and slight to moderate
corneal injury. 

Skin irritation - Rabbit A single short exposure
resulted in slight erythema (minutes), slight
edema, and a slight to moderate burn 

Rabbit - Twenty daily applications of a 10% chloro-_
form solutiOn of the product to the ears of rabbits
on a bioassay test for chloracnegenic activity
resulted in no activity. -

:teaas

OTT 

-5-

<0.02 g,iLo-Trichlorophenol 

 

Acute oral - Rat - male - LDSO-3250 mg/kg body wt. - 
Lethality - Rat - female - LD50-2830 mg/kg body wt.

bye Irritation Rabbit - The product caused moderate
to severe conjunctiral redness and swelling, slight
to moderate iritis, and slight to moderate corneal


.Skin irritation - Rabbit - A single short exposure of
the product on intact skin resulted in slight
erythema (1 hr), and moderate erythema, slight
edema and slight necrosis (4-5 hr).

Prolonged exposure (24 hr) of the product on intact
and abraded skin resulted in moderate to severe
erythema, very slight to slight edema, and a moderate
necrosis (random abraded skin areas).

Rabbit - Twenty daily applications of a 10% chloro-
form solution of the product to the ears of rabbits
on a bioassay test for chloracnegenic activity re?
sulted in no activity.

Inhalation - Human Experience has shown that the
dust is irritating to the nose and throat.

If any of you know of further data; please supply me with copies
of it so I can update this.

The data given is a quick summary. Should more detail be
needed, please let me -



Mark A Wolf

Information Services
Toxicology Research Laboratory
1803 Building

isiaa



bcc: S. B. Sachs
G. E. Lynn??Action File-_

 

KURON
December 30, 1964
Mr. George M. Downard, Head (2) :3
Registration Section (3
Pesticides Regulation Division :5
Agricultural Research Service
U. S. Department of Agriculture to
Washington, D. C. 20250 CD
03

Dear Mr. Downard: 5?


Subject: Proposed use of on Sugar Cane in Florida
(KURON - USDA Registration No. 464?162)

By copy of the attached communication from J. R, Orsenigo,
Associate Horticulturist, University of Florida, Everglades
Experiment Station, dated December 23, 1954, it will be
noted that certain weeds of sugar cane plantings in Florida
are not now controlled with and herbicides;

In some cases the long continued use of has apparently
resulted in development of resistance of the weeds and in
others the weeds have never been susceptible to the use of
and/or 

Several of these weeds, such as the fennels,
ground cherry, purslane and wild lettuce do respond to treat-
ment with silvex, the name of the active ingredient in

.

Since HURON is registered for use in sugar cane plantings in
Louisiana and Hawaii, and since it is effective against the
weeds in question in Florida when used at the same rates and
under similar application conditions, it seems appropriate

to apply for use registration in Florida, as indicated by the
attached proposed addition to the HURON label.

We therefore respectfully apply for registration as indicated
and will appreciate the earliest practicable action by the
Pesticides Regulation Division pertaining thereto.

Very truly yours,

?ag/W

O. H. Hammer 
Registration Section I 
Bioproducts Department

'cc: W. W. Sunderland, Dow Washington Office
I

 165030

.hagg
254G
- C3
AQUATIC VEGETATION CONTROL IN RECREATIONAL AREAS C)
by is
1 to
Mark Wiltse co
pa

CD
CD

Introduction

Aquatic weed growth has become an increasing problem in recreational
areas of the eastern half of the United States. Nature does abhor a
vacuum in the plant world,and in aquatic areas of shallow water this 13
especially true. A succession of biological organisms and algal growth
followed by the invasion of aquatic weeds, emergent aquatic growth, and
higher forms of emerged aquatic vegetation and terrestrial plants, is
nature's way of converting aquatic areas to dry land.

Aquatic vegetation has restricted utilization of water for such purposes
as hunting, fishing, bathing, water skiing, boating, and in seme
instances has destroyed the normal beauty of aquatic areas. Fish pro-
duction can be limited by excessive growth of water weeds. Aquatic
vegetation is often a problem in water areas with limited water movement.
Man has been trying to interrupt this evolution of aquatic areas for
some time, in-order to maintain the aquatic or water areas so they can
be of maximum use to man.

Aquatic vegetation control mEasures have been used for many years and
haveconsisted primarily of mechanical techniques. These contrivances
have ranged from underwater cutting bars to hand rakes. Although these
mechanical methods are often effective for a short time, regrowth from
roots or rooting of stem fragments often increases the problem. The
control of water levels or draining and reflooding water areas, has also
been used to some degree. These methods have generally been laborious
and provide only temporary relief. In recent years, chemical control
methods have offered a new opportunity in aquatic vegetation control.
In discussions on aquatic vegetation, it is desirable to consider the
various aquatic weeds, depending upon their habitat and type of growth,
since the control measures for the various groups differ considerably.
For this discussion, let us consider control measures for the following
broad classification of aquatic vegetation-~submerged, floating, and

emergent aquatic weeds.
Control of Submerged Aquatic Weeds

Submerged weeds are probably the major weed problem in recreational waters
or at least causes the greatest concern where the water is used for
bathing, boating, fishing, water skiing, etc. Submerged aquatic Weeds,
such as milfoil, coontail, and pondweeds, can completely fill ponds that
are 7 to 8 feet deep with aquatic growth, so that is impossible to even
-row a boat through the water. Bathers have become.entangled in this type

 

1The Dow Chemical Company, Midland, Michigan 

(Paper presented at 1961 Michigan Forestry and Park Association
Anniull Conftuwnxce. hxust Lansiuu;, Mic?iigan.)

MOO

of growth and drown because of their inability to free themselves. Sub-
merged aquatic weeds also tend to create a fish imbalance by providing cg

protection for the small fingerlings, so.that the large fish cannot co
consume them. This gives rise to a large population of small fish whichF?
consume much of the food supply. Fishermen often have difficulty in :3

getting their hook and line through these weed masses. ?1

The New Jersey Fisheries Laboratory at Milltown, New Jersey, conducted
extensive tests on this problem and in 1956 demonstrated the activity of

silvex for the control of submerged aquatic weeds (6). In their test,
they found that silvex was effective on a broad spectrum of aquatic
weeds using 2 of the active ingredient. Since 1956, silvex has been

tested and used commercially for submerged aquatic weed control thrOugh-
out the United States. In 1958, Kuron* herbicide, Dow's propylene
glycol butyl ether ester formulation of silvex, was registered for this
use by the Pesticides Regulation Branch of the United States Department
of Agriculture. Kuron, when used at the rate of 5 quarts per acre feet,
will control certain submerged aquatic weeds such as water milfoil,
fanwort, bladderwort, coontail and waterweed. Kuron should be applied
in early summer when the water temperature is and weeds are
approaching the water surface. To apply preperly, first dilute the
Kuron with enough water for uniform distribution over the water surface
with the available equipment. Kuron has been especially effective when
applied to water areas that have a minimum amount of water movement.
Ponds with some water movement can be successfully treated if the water
level can be lowered so there will be no overflow for at least three
days after treatment. This rate of application of Kuron will also give
control of some of the floating and emergent aquatic weeds, such as
water lilies and spatterdock. Applications of Kuron ordinarily have not
been harmful to fish; however, in certain situations where complete pond
treatments have been made and where concentrations higher than the

5 quarts per acre foot have been used, some fish kill has been observed.
Recent work in Massachusetts (1) has indicated that lower rates of Kuron
have been effective on such weeds as water milfoil and bladderwort, and
they are using 1-2 quarts per acre foot of water in some of their lakes.
Kuron has a low mammalian toxicity, and normal precautions used in
handling agricultural chemicals is sufficient for the use of this
'material. The Kuron formulation of silvex is an ester and an emulsion
is formed when it is sprayed into the water. -

Our company has been actively evaluating other formulations of silvex
for aquatic vegetation control and has developed a salt formulation
which forms a true solution with water. This has given very encouraging
results;~ Silvex salt formulations will possibly be available in test
market areas in Michigan this season as'KurosaI*G? and "KurosaI*SLW.
Kurosal contains 2 silvex on a clay granule, while Kurosal SL
contains 6 pounds of silvex per gallon. Kurosal should be applied at
100 to 150 pounds per surface acre, depending upon the depth of'the -

?water. Kurosal SL is designed to be applied asua direct application
-nto the water surface by a bOOm or by injecting a stream of the chemical
in the wash of an outboard motor. Kurosal formulations appear

paniicularly safe to use in waters inhabited by fish.

issaz

 

a:
Trademark of The Dow Chemical Company

Silvex is the most promising deve10pment in the aquatic herbicide field
and offers the potential of solving many of the submerged and emergent

aquatic weed control problems without the necessity of using materials

which are toxic to animals.



is available both as an ester and an amine salt and has been used
to some extent for submerged aquatic vegetation control. In 1956,

Dr. Buford Grigsby, from Michigan State University, reported on his .
results using a granular carrier and there has been considerable
interest in the use of granular for submerged aquatic vegetation 
control since that time. Esteron* 99 Granules herbicide,which contains
20% as the propylene glycol butyl ether ester on a clay granule,

is one of several available commercial formulations of granular 
granules applied to the surface of the water by hand or mechanical
devices sink rapidly to the bottom of the water releasing It was
originally thought that adhered to the granule for long periods,
releasing slowly and thus localizing the active ingredient around the
roots of the submerged aquatic weeds. Some recent work has indicated
that actually is removed from the granule fairly rapidly, and that
it is dissolved into the water. However, the use of the granule carriers
for distribution purposes is convenient for small scale applications
around boat docks and in front of cottages, etc. Although some workers
have indicated that granular was effective, regardless of water
lepth, it has become evident from later work that water depth is quite
important to obtain satisfactory control. To insure effectiveness,

should be applied at about 5 pounds acid equivalent per acre foot
of water treated. Water movement may also decrease the effectiveness

of granules, since the active ingredient can be removed from the
site of application before it is active on the plant.

Esteron 99 Granules or other granules containing 20% should
be applied at 100 pounds.per surface acre in water that is 4 feet deep
with an additiOnal 25 pounds per surface acre for each foot of depth in
excess of 4 feet. granules must be applied early in the growing

season to be effective.

Later season treatments on weeds not actively growing have not been
consistently effective. granules will control such weeds as water
milfoil, coontail, and bladderwort, but is considerably less effective
on pondweed (Potamogeton spp.). granules are not as effective on
as broad a spectrum of weeds as sodium arsenite or silvex. In the
granular form, is easy for the boat owner or cottage owner to use

in Small areas.

Control of Floating Weeds

Water lilies are a problem in recreational waters since they often .

?interfere with boating, water skiing, and swimming. Water lilies can
ecome so dense that it is nearly impossible to get through the area with

a boat. Water lilies can be most effectively controlled by surface sprays

with silvex. We recommend the use of Kuron at 2 quarts per 100 gallons

"am" 18503

 

 

Trademark of The Dow Chemical Company

of water to be sprayed as a wetting Spray on the water lilies_when they
are in an active growth period, and the leaves have fully developed.
Late season applications when the water lilies have completed their
vegetative growth have not been consistently effective. esters
have also been effective for the control of water lilies; however, the
results have not been as consistent as with applications of Kuron.
Duckweed (Lemna Spp.) is a free-floating aquatic weed and is often a
nuisance in sheltered areas and may c0mpletely cover the water surface.
Duckweed has been very troublesome to control, since it can reinfest
areas very rapidly. It has been reported that several materials will
give immediate kill of the duckweed but reinfestation within three or
four weeks is common.

some MOO

Control of Emergent Aquatic Weeds

 

Emergent aquatic weeds have been a severe problem in waterfowl marshes
and have decreased the waterfowl populations in some areas. Phragmites,
cattails, and other weeds and grasses can cover an entire marsh,and thus
open areas are not available for geese or ducks to land, and nesting and
feeding areas are not attractive to them. Of course, emergent aquatic
weeds are also a problem along the shoreline of recreational waters,
cottages, etc. Cattails and other narrow-leafed plants can be controlled
effectively by using Sprays of Dowpon* or Radapon*. Radapon at 10 pounds
per 100 gallons of water should be applied as a wetting spray when the
weeds are in an active stage of growth. Aerial applications of Radapon
at 10-15 pounds per acre in 5?10 gallons of water have been very effec?
tive for the control of cattails for opening up marsh areas. Radapon is
particularly effective on narrow?leafed plants and has little effect on
many broad-leafed plants such as smartweed, pokeweed, wild buckwheat,
etc., which are desirable for waterfowl food. Radapon is less toxic

than common table salt and has been used in water reservoirs for aquatic
vegetation control without causing any deleterious effect on water

quality.

Broadleaf emerged weeds often grow along the shoreline and may be a
problem in some recreational areas. Some of these problem weeds are
pickerelweed, arrowhead, water plantain, etc. Spray applications of
Kuron using 2 quarts per 100 gallons of water have been effective in
control of these weeds. Applications should be applied when the plants
are in an active stage of growth. Emergent aquatic vegetation consist-
ing of both narrow-leafed plants and broad-leafed plants can be effec-
tively controlled by a mixture of silvex and dalapon which is sold
Icommercially under the trademark, Garlon*. Garlon, used at 3 gallons
per 100 gallons of water, has given effective control of cattails,
burreed, arrowhead, pickerelweed, etc. Applications.should be applied
when the plants are in an active growing stage to be most effective.



 

Trademark of The Dow Chemical Company 

165M.

-5-

Summary

EMOG

Several herbicides are now available for the control of aquatic weeds
and algae in recreational areas. New herbicides have shown some promi 8?
for the control of submerged aquatic weeds and additional research wil
probably develop new chemicals to be useful in this field. There are as
several chemicals presently available that can do a very effective job c:
in the control of aquatic weeds and should be very useful in the
management of recreational waters and marshes.

16505;

. . - 

-5-

 

Literature Reviewed
C3
2
1. Cortell, J. M. Re-evaluation of the Concentration Required fangs
Effective Aquatic Weed Control with Silvex. to
PROCEEDINGS OF NORTHEASTERN WEED CONTROL CONFERENCE h?
14:478-482, 1960. he
. h+
h.

Aerial Control of Cattail with

2. Heath, G. and L.4C. Ruch.
Radapon. DOWN TO EARTH, Winter 1957 Issue.

The Chemical Control of Aquatic Nuisances.

3. Mackenthun, K. M.
Madison, Wisconsin,

Committee on Water Pollution,
January 1958.

4. Palmer, C. M. Algae in Water Supplies. U. S. Department of
Health, Education and Welfare. Public Health Service

Publication No. 657, 1959.

5. Wiltse, M. G. Aquatic Vegetation Control with Kuron.
(Presented at 1960 Meeting of Weed Society of America,

Denver, Colorado.)

6. Younger, R. R. A Preliminary Report On Controlling Aquatic
Vegetation in New Jersey with Kuron. DOWN TO EARTH,

Spring 1958 Issue.

16503

 ACD FILE NO. 
 AQUATIC VEGETATION CONTROL WITH meow 2547
by

C3

2
-. 
. Mark Wiltse to


0
ED

AQuatic weed growth has become an increaSing problem in recreational water
of the eastern half of the United States. Vegetation has restricted
utilization of waters for such purposes as bathing, water skiing, boating,
and in some instances has destroyed the normal beauty of aquatic areas. 
Fish production has often been limited by excessive growth of water weeds.
A number of control methods have been employed with varying degrees of
success.

Mechanical contrivances have ranged from under water cutting bars to hand
rakes. Although these mechanical methods are often effective for a short
time, regrowth from roots or rooting of stem fragments often increases the
problem. The method is generally laborious and provides only temporary
relief. .

Sodium arsenit? has been used as a herbicide for several years. In some
areas, particularly static waters, this well-known chemical has given out-
standing results. Principal limitations to its use have been its inherent
to 'city, resistance and failure to control a broad range
of .egetation.

Several workers have been actively evaluating products which would control
submerged aquatic weeds and overcome the limitations of previously existing
methods. The New Jersey Division of Fish and Game inaugurated a five?year
research program in 1951 financed by Dingell-Johnson funds to find an 
iquatic herbicide (8). In the course of this work, Kuron herbicide was
found adaptable to this problem and seemed to offer control comparable to
sodium arsenite (8). Kuron contains the-active ingredient silvex
propionic acid as the propylene glycol butyl
:ther ester and acts as a plant growth regulator.

Aquatic Weeds Controlled

 

Early tests indicated that Kuron herbicide at 5 quarts per acre foot would
Control many of the troublesome submerged aquatic weeds in the northeastern
United States Experimental tests in 195? and 1958 in several
sections of the country evaluated the effectiveness of this concentration
on several aquatic weeds. Some tests were conducted at reduced concen-
trations by certain workers (2). Commercial applications in 1958 and-1959
iontributcd additional performance information for controlling certain

I
I

1" rademark of The Dow Chemical Company

1' The Dow Chemical Company, 916 Shoreham Building,
Washington 5, D. C. . 

16507

?Papcr presented at 1960 Meetin of the . "i 
Denver, Colorado.) 6 cad or ?merica'



species. The following list summarizes some of the weeds controlled by

 

C3
Kuron. . g;
Agustic Weeds Controlled by Kuron at l.2?1 2.5 and 5 Quarts per Acre Foot

. CD

1.25 quarts/acre foot or less (0.5 ppm) EB

White water lily app.) . 
Pickerelweed [Pontederia cordata L.) h?

 

2.5 quarts/acre foot or less (1.0 ppm)

 

Yellow water lily . . iNuphar spp.) 
Mud plantain . . Heteranthera spp.)'
'Water milfoil 

 

5 quarts/acre foot or less (2 ppm)

 

 

 

Bladderwort - -. Utricularia spp.)
Watershield Brasenia schreberi)
Coontail demersum)
Fanwort . {Cabomba caroliniana)
Haterweed . LAnacharis canadensis)

 

Potamogeton diversifolius)
Potamogeton amolifolius)

Variable pondweed 
N, 1d_ lNaja: flexilis)
. {Eleocharis spp.)


 

 

 

 

Sp-ge rush 

Tapegrass - . Vallisneria americana)
Haterstarwort Callitriche sp .)
Soft rush Juncus effusus
Bulrush -- . -LScirpus americanus)

 

Burreed - (Sparganium spp.)

In some treatments, these weeds were controlled at lower concentrations,
but there were not sufficient tests to indicate consistent effectiveness

at the lower rates. Treated areas observed two years after treatment
indicated a continued control over this period of time. In total treat-
ments on ponds and lakes with Kuron at 5 quarts/acre foot two years control
has been obtained in some instances.

Application Technique

 

Emcrged species were controlled in many instances by using Kuron at one
gallon per 100 gallons of spray solution applied to the emerged foliage
as a thorough wetting spray. The applications for the control of the sub-
merged aquatic weeds were generally applied by diluting the Kuron at one_
gallon in 10 to ?0 gallons of water and directing this solution on or?inio
.the water. Proportioning pumps were often used to mix Kuron with lake
.w :r and pump it to the boom which would distribute the solution over a
wide swath. Uniform distribution appeared to be desirable to allow a
lethal concentration of the Kuron to come in immediate contact with all
_the vegetative growth. 
- 16508

Injection of the undiluted formulation of Kuron was used in some equipment,
but on a limited basis. . I .

Environmental Effects



Applications of Kuron appeared to be most effective if treatments were
nade during periods of active growth. Treatments applied when water I
temperatures have been or higher have been most effective. The watet?
Itemperature may not influence the reaction of the chemical to a significant
degree, but the aquatic weeds may be in their most active growth at thesecg
higher temperatures and thus more susceptible. Undoubtedly, the most b3
active growth periods vary for the various aquatic weed species.

applications with Kuron have been most effective on submerged aquatic 
plants in lakes and ponds where there is no or limited water movement.

Less satisfactory control of some plants has been observed where water cur-
rents replace the treated water soon after application. Translocation has
>een demonstrated in water lilies where applications to the leaf surface
1as caused a herbicidal effect on the rootstock. However, some aquatic
)lants have only primitive vascular systems which may not be functional

1nd translocation to root areas probably is not important. In these plants,
1 lethal concentration of Kuron may be required to contact all the vegeta-
;ive portions of the plant which are capable of?growth or reproduction.
11though the exact minimum contact or exposure time required is not clearly
iefined, MB to 72 hours exposure to treated water has given control.

Pru slut and Williams (7) compared the effectiveness of Korea for control-
iing coontail demersum).in greenhouse-tank experiments at
;hree pH levels and three concentrations of calcium ions. Kuron was most
:ffective on coontail growing in acid water. It was also more
Lctive on coontail growing in water with 250 and 500 of calcium ion
:han in water with no calcium. In these tests, the effect of calcium was
lot differentiated from a possible effect of chloride ion which was added?
.n equivalent amounts. -Field plot treatments and commercial applications
rave been only better under acid water-conditions compared with
~reatments to neutral or alkaline water. Perhaps the water pH is
unly important in water situations with a low level of calcium ions or
iimilar dissolved ions. 

 

afety to Fish and Other Aquatic Organisms.

 

'he experimental treatments to plots and complete ponds have caused no fish
nJury at concentrations of Kuron of 1.25 to 5 quarts per acre foot.
ommurcial_applications at concentrations of 5 quarts per acre foot or less'
ave given no appreciable fish ki?.in numerous treatments throughout the
ountry. Slight fish kill has occurred in some'instances where applications
ere made at higher concentrations. The higher concentrations sometimes
ccurred in shallow areas where the spraying equipment could not operate
reperly and thus higher concentrations were developed. There has been no
isk??njury in over 99% of the treatments applied; ?apid incorporation of
he -prsy solution particularly_those associated with rates in excess of
quarts per acre foot of Kuron should be avoided where fish kill cannot be 

elevated. - . .
16509



Pierce found that applications of Kuron had no influence on oxygen
content in treated plots. Aquatic weeds treated with Kuron normally 

dr ?mpose slowly, in most situations 10 to 21 days are required for decom-
pssition. This slow decomposition apparently does not lower the oxygen
content of the water.. This is a problem with some aquatic herbicides which
give rapid kill of weeds.. Pierce also reported that Kuron treatments showed
no effect on population of benthic organisms represented by the following
groups: Annelida, mostly leeches; Mollusca, including seven species of
Gastropoda and one Specie of Pelecypoda (Sohaerium); Amphipoda; Insecta,
larvae of mayflies, damsel flies, dragon flies, midges and beetles.
Applications of Kuron temporarily decreased populations of some plankton,
but within two weeks after treatment the populations were equal to
untreated areas. These plankton consisted of fifty identified species in
the following large groups; Myxophyeeae, Chlorophyceae, Desmidaceae,
Diatomaceae, Flagellata, other Protozoa, Nematode, Rotifera, Annelida,
Crustacea. There was no deleterious effect on large aquatic vertebrates
such as frogs and turtles in the treated areas. -

801188 MOO

Prolific algae growth is often a problem soon after treating aquatic weeds
with some aquatic herbicides. Algae growth has not normally occurred until
three to four weeks after treatment with Kuron. The delay in appearance

of algae apparently is associated with the slow kill of the aquatic weeds
and thus the delay in release of nutrients into.the.water from the decaying
vegetation. 

14fcty to User
{uron herbicide is low in toxicity and presents a_low degree of hazard in
landling and use. . -

Ihen Kuron herbicide was fed by intubation to groups of various animal
species, the values were found to be as follows: rat, 1070 mg/kg;
guinea pig, 850?mg/kg; rabbit, 850 mg/kg; mouse, 2140 mg/kg and chicks,
3000 mg/kg. From these data, it may be concluded that there should be no
iroblem from ingestion incidental to the handling and use of this product.
LE large quantities are swallowed, accidentally or willfully, some injury
light result. Hdwever, the likelihodd cg serious?inJury_Is remote.
Ievertheless, Kuron.herbicide should not be left where children or live-
;tock have access to it. - 

 

 

 

 

iattle, sheep, swine, ducks and chickens have consumed water containing
.uron herbicide at 50 over a four week period with no adverse effects
a determined by water consumption, body weight gains, and gross observa?
ions, including gestation and parturition in sheep. These data indicate
hat foliage or water treated with Kuron herbicide, as recommended, should
ot present a hazard from ingestion. -

kin irritation tests conducted Upon rabbits have indicated that Kuron

crbieide and its dilutions are irritating, upon repeated pro?r
onged contact. In these studies, there was no evidence of absorption 
hr 3h the skin in acutely toxic amounts. The observation of reasonable
are and personal cleanliness practices should be adequate to avoid skin

ifficultic?. - - 
- 16510

 

. 

Kuron herbicide, when diluted to 1% with water, caused no primary irrita~
tion nor allergenic responses when applied to the skin of fifty human
volunteers using a repeated insult technique. One subject did develop a

?1d fatiguing response. In addition, the product was "patch tested? on a
Second grOUp of fifty human subjects at concentrations up to 50% without
cutaneous reaction. Even when applied undiluted to the skin for two hours,C3
it caused no skin response. Thus, it may be concluded with a high degree
of assurance that when used as directed, this product should cause no skin
irritation or skin sensitization responses. 

When tested on the eyes of rabbits, Kuron herbicide and its solutions
produced moderate pain but only slight irritation. Thus, this product
presents no unusual hazards from contact. However, suitable pro-
tection, such as safety glasses, may be worn simply to avoid the discomfor?h
that might result from contact. Swimmers should not experience any discom-
fort from water treated at recommended concentrations.

01188

Effect on Water Use

 

Kuron may remain in water for three to four months after treatment where
no dilution occurs. An 88?acre lake was treated in New York after the
'water level was lowered two feet. No flow occurred through the outlet
until three months after treatment. In analysis of the lake water thirty
days after treatment, 2.4 of silvex from Kuron was present, essentially
the same amount that was applied. Analysis of the lake water seventy days
after treatment showed 2.1 of silvex present. Samples of water
analyzed 100 days after treatment and thereafter showed no Kuren present-
Ch 'icai analysis of the bottom mud indicated no build?up of Kuron. Since
Ku.an herbicide is an organic.compound, the components would be expected
to be bacteriologically decomposed principally to the elements carbon,
hYdrogen and oxygen. 

Silvex acts as a growth regulating compound on terrestrial broadleafed
plants and is used as a stop?drop treatment prior to harvest of apples at
concentrations of 10 to 20 ppm. Although 2 of silvex as used for
aquatic weed control might not cause inJury if sprayed on some broadleafed
plants, some susceptible species may be affected. Accordingly, it is
suggested that water treated with Kuron should not be used for spray pur-
poses or for irrigation during the season of treatment until more informa-
tion is available on the possible hazards to crops.

Summa ry

:urdn aquatic herbicide is effective on several submerged and emerged
Lquatic weeds. Applications to still water at 5 quarts per acre feet are
iuggested for general aquatic weed control. Lower rates are effective on
ertain species. Conditions for growth, pH and dissolved ion content of
'aLer may influence the effectiveness of Kuron. The low mammalian toxicity,
low action, and safety on fish food organisms are desirable preperties-of
uron aquatic herbicide. 

-. - 

-. . '16511?_

 

- . . I 

Literature Cited

1. M. M., "Field Testing of Kuron as an Aquatic Herbicide
in Massachusetts," Proc. NENCC, 13:315-321. 1959

2. J. M., "Re-Evaluation of the Concentrations Required for
Effective Aquatic Weed Control with Silvex", Proc. NENCC
14:h78?H82. 1960



3. Hall, wt 0., "Control of Various Aquatic weeds with Silvex",
Proc. NEWCC 14:476?477. 1960 -

4. Pierce, M. E., "The Effect of the Weedicide Kuron upon the Plora
'and Fauna of Two Experimental Areas of Long Pond, Dutchess
.County, New York?, Proc. NEWCC 12:338-343. 1958

 

'"Further Studies of the Effect of Kuron upon the

Flora and Fauna of Long Pond, Dutchess County, New York",
Proc. HENCC 135310-314. 1959?

i 6. "Progress Report of the Effect of Kuron upon the

Biota of Long Pond, Dutchess County, New York?, Proc. NEWCC
1960 

I

7. Truchelut, G. E., and Williams, h. C., "Some Observations on the

Influence of Dissolved Ions on the'Effectiveness of
Phenoxy-Type Herbicides on Submersed Aquatic Weeds", Proc.
SWCC 13:2Elr227. 1960 . 

8. Younger, R. R., Preliminary Report on Controlling Aquatic

Vegetation in NeW'Jersey with Kuron", DOWN TO EARTH,
Spring?l958. . .

9. Younger, R. E., "Progress Report on the Use of Kuron, and

Granules on Aquatic Herbicides", Proc. NEwcc -
13:322-329. 1959 -

16512

A.

 

 

KURON silvex herbicide
continues to gain acceptance

By Lawrence Southwick
Dow Chemical U.S.A.

For more than two decades, three chemicals
in the phenoxy group of systemic herbicides.
i.e. 2.4-D. 2.4.5-T and MCPA. have more Or
less dominated the attention of researchers.
advisors. marketers and users. Generally
speaking. 2.4-0 and MCPA are still leaders
in broadleaf herbaceous weed control. and
2.4.5-T. alone and in combination with
2.4-0, is widely used in wcody plant control.
The purpose of this brief paper is to review
some of the herbicidal possibilities of silvex.
This report does not and is not intended to
encompass an exhaustive survey of the con?
siderable literature generated over the pasr
20 years relative to this herbicide. Selected
published papers are referenced to provide
support for the thesis that silvex is indeed
a very effective herbicide with a broad use
spectrum.

Historical Background

Shortly following the introduction of 2.4-0
in 1945. researchers began to characterize
its herbicidal utility in several use areas. It
rapidly found acceptance for controlling
herbaceous broadleaf weeds in turf. small
grains. corn and in non-crapland. It also
showed promise for controlling woody
species. However. as early as 1946. it be-
came obvious that 2.4-D was not adequately

effective on certain weeds. for example.

chickweed in turf. and on many woody
plants in non-cropland brush control.

By 1947. Dow research workers in Michi-
gan. workers in Hawaii, and probably others
reported signiticant differential species re-
sponse to 2.4-0 and 2.4.5-T. The latter soon
found a place in the control of woody plants
and also in selective weed control in rice.
The idea of combining these two herbicides
to broaden the Spectrum of woody plant

26 Down 1'0 mam. Vol. 23. Nu 4. Spring 1973

NJ

pl;-


ZSOIEB Moo

control was put into practice. Commc
spraying of right?of-ways with mixtUn
12.4-D and 2.4.5-T became a general pra
and such combinations are in wide u:
the present time.

Early Dow studies in Michigan (1952
dicated that another related but chemi
Specific herbicide. 
propicnic acid was. very effective on a 
assertment of herbaceous weeds and Wt
plantsz. Silvex was coined as the com
name. herbicide. a low voi
ester formulation of silvex. was firSt i
keted in 1954.

Beginning around 1954. many rep
have been presented at the regional v.
control conferences on the herbicidal 
acteristics of silvex. For example. in Ml
gan trials. it was reported very effectiv:
four species of oaks. and on mapleJ. In
tensive tests in res

. checked three years after application sho

silvex effective "?on mixed oaks and a;
ciated species?. In Oklahoma. silvex
excellent on "all oaks" which included 53-.
white. blackjack. black and chinquap
Based on many field trials. silvex was fo
to be an excellent overall turf herbicide
controlling a wide assortment of weeds
eluding many that are tolerant to 
an aquatic herbicide. silvex was reporte'
be superior to others-in Oklahoma?.
northeast" and elsewhere.

An early repon on basal treatment 
KURON herbicide was made In 1956 on .
key oak in Florida?. Summer 

were rated excellent while Winter treatrr

was moderately effective. Work in Ore:
proved the effectiveness of KURON in a.
basal control of big leaf maple?. Curr

16513

 


a



.
Anni-nah.qu-

?4-1414 @mu?k?m.g?h

 

LITERATURE CITED

(1) Arend. J. 1967. Some variables affecting
hardwood control with stem injections of
North Central Forest Experiment
Station. USDA Forest Research Paper NC-
18. 11 pp.

(2) Brinkman. K. A 1970. Picloram in spaced
stem injections to control Lake States hard-
woods. Nonh Central Forest Experiment Sta-
tr0n. USDA. Forest Research Note NC-94.
4 pp.

l3)Carvel. K. and F. C. Cech. 1967. if used
properly -tree injectors are effective! Science
Serves Your Farm and Home. Agricultural
Experiment Station. West Virginia Univer-
srty Bulletin 552: 6-10.

l4) Cope. J. A. and J. N. Spaeth. 1931. The kill-
ing of trees with sodium arsenite. J. Forestry
29. 775-783.

l5) Nation. H. A. 1967. Report on tree control
via injection with Tordon 101 mixture. Down
to Earth. Number 2: 24-27.

(51 Norris. L. A. 1971. Chemical brush control:
assessing the hazard. J. Forestry 69: 715-720.

Hy'ker. R. A. and L. S. Minckler. 1962. Methods
and costs of killing hardwood culls. Central
States Forest Experiment Station. U.S.D.A
Forest Service Technical Paper 191. 9 pp.

r8) R. D. 1966. Soil-applied urea herbi-
cides in the control of unwanted wood-,-
?hulls Aurrcultural Science Review. First
Quarter 1966. Vol. 4. No. 1: 21-27.

(9) 1968. Penetration and ac~
cumulation of fenuron in forest soils. J.
Forestry 66: 910-912.

(10) Shipman. R. D. 1969. Pelleted vs. liquid
herbicides for control of low-grade white
oak in central Proceedings.
Northeastern Weed Control Conference. Vol.
23: 298-302.

. 1969. Establishment and
early release of underplanted Japanese larch
with soil-applied fenuron. Tree Planter's
Notes. Vol. 20. No. 1: 23-27.

(12) 1970. Some variables af-
fecting the response of unwanted hardwoods
with tree-injected Proceedings.
Northeastern Weed Control Conference. Vol.
24: 250-256.

(13) Smith. J. 1959. Tests of injected chem-
icals for hardwood control in the Arkansas
mountains. Southern Weed Conference.
Shreveport. L0uisiana. 3 pp.

(14) Sterrett. J. P. 1967. injector treatments on
oaks. red maple and hickory with several
water soluble herbicides. Proceedings.
Scuthern Weed Conference. Vol. 20: 190
(abstract).

(15) Watson. A J. and R. J. Mesler. 1964. Effect
of Tordon herbicide as basal frill and tree ins
jection treatments on certain hardwood trees.

(16) Down to Earth. Vol. 19(4): 20-23.

Wiant. H. V. and C. Walker. 1964. Com-
parison of two silvicide injectors for killing
oaks. The Consultant. Vol. 9, No. 4: 16.

(11)

 

te-

860188 

DOWN TO EARTH. Vol. 23. No 4. Sunni) 197]

25.

CI
Cl

Oregon recommendations include silvex for
. this species as well as for vine maple Dor-
mant treatments on salt cedar in New
Mexico were moderately effective?. Com-
mercial use of basal application of silvex has

corroborated effectiveness on many species
of woody plants. An effect commonly n0ted
is delayed maximum response. Final control
has been satisfactory.

Extensive aerial trials in Hawaii showed
KURON herbicide to be the most promising
iof those tested to reclaim wetland brush
 jungle for cattle range?. Weedy species in-
3 cluded melastoma. false staghorn fern. ohia.

and lantana. Work in Oregon proved the
 superiority of silvex in air or ground applied
lfoliage sprays on dalmatian toadflax and
i this is the present state recommendation in
Oregon? as well as in Montana. Control of
 prickly pear cactus with silvex is recom-
mended in Montana. Nebraska. Wyoming
and elsewhere. Data in the Research Prog-
ress Reports of the Western Weed Control
Conference. even prior to 1962. indicated
good control with KURON on several diffi-
cult species. including cholla cactus. com?
mon tansy, dalmation toadflax. iantana.
larkspur. mesquite, milkweed. perennial
peppergrass, poison oak. prickly pear cac-
Htus. Russian knapweed. salt cedar. snow

1: berry. sorrel. turbinella oak. whitethorn.

wild rose. yucca and others. For many years.
. i silvex has been used extensively in turf and
sugar cane and for aquatic weed control.

.I Weed and Brush Control with KURON

Two excellent references which charac-
terize comparative effectiveness of phenoxy
herbicides are: USDA Farmers' Bulletin No.
2183 (Jan. 1971 revision) "Using Phenoxy
Herbicides Effectively"". and USDA Agricul-

A. LII. wanna-AL



-.-- --



4954?!

he:

2 (2,4,5-trichlorophenoxy) propicnr'c acid

ture Handbook No. 269 (March 1955) ?Her-
bicide Manual for Non-Cropland Weeds?.
Tables 1 to 8 were developed largely from
these information sources.

Since the effectiveness of silvex has not
been adequately reported on many species.

. the listings in the tables cannot be con-

sidered "complete". In Tables 1 through 4.
effectiveness is given in terms of excellent.
good. or fair which are charaCterized as
follows:
excellent-one application at a normal
rate (1 lb/aCre) kills the plant in most
cases.
good?more than one application at the
normal rate or an application at a higher
rate (2 to 4 lb/acre for weeds and 4 to
8 lb/acre for woody plants) needed for
control. -
fair?repeated applications at the nor-
mal or a higher rate often required for
adequate control.
it is interesting to note in these tables the
wide range of annual. biennial and peren-
nial weeds. woody plants and aquatic weeds
that are controlled with silvex. This relatively
old, established herbicide obviously has at-
tributes that may not have been fully eval-
uated and certainly were not fully developed
due. in part. to the diversion of interest and
research effort to the great influx of new
families of herbicides which emanated from
the research laboratories coincident with and
shortly following the initial development of
silvex. Yet the limited research studies with
KURON herbicide and other silvex formula-
tions as well as successful commercial
usage have delineated specific use? areas
wherein the special attributes of silvex her-
bicide have become recognized. The infor-

 MOO

Down TO EARTH. Vol 23. No. 4. Sprint. 1971 27

16515 



Control of Annual and Biennial Weeds with Silver

 

 



unwind:

huggam'clzs
binlmud
?lo-mun!
butclonr
burdock

humlcups
:hictweed. can.
coiluhln
canon
doglonnll
dulsalu
anningpu'msosl
liddlonldi
llnhanu
gllinsugl
groundcheny
halo-mustard
hemlock. poison
lnawsl. annual
lochia
lamhsquamrs
marsh-Ida:
morningglory
mustard:
partridge?:
passionllower
liald
pappcmeods
pigwuds
phnlaim
poisonhemloch
radish, wild
Iagwuds

llpl. him

ushania (colluheanl
smouweed
spanishnudlos
sunllowu
thistll. bull. Russian
liluino

mulch. wild
witchwud
woodsoual. nllow

wmmsud-musmd

28 DOWN TO mam. Vol. 23. No 4. 5mm 1973

GOOD

binomial-d
tolls-uni
lioldpepoargrlu
guanium. 
goosoiool. oaklul
gtoundsol. ausluf
hlnhit

horsowud (muesli!)
knapweod, spo?ad
lnolwud, iapanoso
medic, blank
uninha-
moxicanwud
mudplamail
Nut
ooh-wold

punlam

rocle londnu
rock-L yellow
puls-
smutwaeds

FAIL

buciwhut t.
?climbs-l. in
?not. will
cmlyindigo


960186 MOO

knolwood. on.
mafwnd (ch;
mullail
poorjoo
punawuino
spur". can



 

 

. L. -. - . -.

TABLE 4. Control ol aquatic weeds with Silvex



 

EXCELLENT
arrowhead 
bladdlmort swamplaoseslriln
toenail 
ultich mtuhyacimh
lanwon unarlily
Innsutrill watermilluil
lotus 
mull-rush 
patron-athei- 
picker-lined wet-maer
primrosnwillow Canada
rush wildcallry

mation in the tables is simply indicative of
the acknowledged wide spectrum of species
response to silvex. An exhaustive survey of
literature and progress reports. and es-
pecially a compilation of user experiences
undoubtedly would reveal additional sus-
ceptible species.

Comparison with 

In Tables 5 and 6. two lists of weed and
woody plant species are given to illustrate
differential foliar responses to silvex and
to These ratings are taken from the
information provided in the two perform-
ance references referred to above?-? with
assists from other data including California
Bulletin 812.? in general. silvex often is
considered mere effective on the species
listed thereunder in the tables and 2.4.5-T
for those listed under it. However. this
does not mean that control of some species
listed under silvex is not possible With
2.4.5-T or vice versa. It simply means that
reported field results on these partiCular
species give the edge to one or the other
herbicide. The differences In ellectivity vary
from moderate to very sngniticant. However.
it should be emphasued that orher field

30 oowu TO EARTH. Vol 23. No 4. Spring 1973

8000



alligatnnvutl .


hurried
entail:


980188 A

data may not be in total agreement w.
that reported here. and likewise many 
sired comparisons have not been reporte
Considering the proven utility of combir
tion treatments of 2.4.5-T with 2.4-D
with picloram for broad Spectrum and i
proved woody plant control. it is likely tl
silvex plus 2.4-D or picloram will be elte
tive combinations and commercial triz

'indicate that this is so.

Comparison with 

In Tables 7 and 8. similar weed and WOOL
plant lists are shown relative to silvex ve
sus 2.4-D. again using information from tr
same published sources. Here again it
obvious that there is a place for bOth 
cides since they do have certain diflerenti
spectra of activity.

it is common knowledge that many herb
ceous weeds and woody plants respOr
quite similarly to most of the phenoxy he
bicides. including silvers. 2.4.5-T and 2.4-l
On the other hand. it is true th
the response of many specnes to these 
chemicals varies significantly. which umph
si:es the need for each ol lhum lht: hurt
cide arsenal. It also emphasizes the pose

16517.

TABLE 2. Conlrul ol Perennial Weeds with Silver:

 

 

ma 

 

t-u- Hun-m )Wl? 

 

u-

 

EXCELLENT GOOD 0

1 chicory hindwud. rim 

cinquoloil .

broomwud docks daisy 

humgc groundivy 

hullucup Ind:- larkspur. duncIcapH

cmcar labour. ull lonuco. hluo 
1 chiciwccd. moon-oar locowocd. whim shunkcahbago 

millwuds sorrcl - 

coyolillo pokownd soudook

dondolioo ago. crooning :omhistlu

dcsorlpanlly chanson-l spikcrush

lirownd mallow-ed. lhrcadlcaf spugos

lupinl. silnry punt-an. wild

milkvotch Viol-1 

mulesnr: "hilltop thistle. Canada

norcalhoan yanow, common

ponnywon. lawn yollowroclrot

ponyioot (dichondrl)

ulch

watuhomlock. spotted

whiteclovor

woodsorrcl

I
TABLE 3. Control of woody plants with Silvax
EXCELLENT GOOD FAIR

alder bololdor wild

upon hunonhusll chon?y. wild

chinam dogwood chokccherry

chollacactuc honoysuchlo crcosololmsh

mplo: locust. black Iln

oak. post mosquito grout-cherry. clammy

oak. shinnary oak. turioy gum. black 8: sweet

 

palmnlo. saw
pucimmon. Tm:
poisonin
poisonoal
pricilypoarcactus
rodbud

IDSI. choroku
ulmonhorry

willowwud

pnnimrnon. common
rasphorry. wild
ton. McCartney
sag-brush. sand
altcodar

aural:
tru-ol-hcavon
willow!

hickory
numaniu
mulberry

oak. hlackiach
oak. hluciach
oak. scrub
oak. whit-
ooaglonngo
mo. roulrillora
sagohlush, big
Iuh'ush

onon
whilelhom

ycrha-sama

yucca [soapwocdl

DOWN EARTH16518







A- .4.



?m ?fLA-n 

1 

 

i

bility of herbicide rotation to forestall the 

development of weed spectra tolerant to a

single phenoxy weed killer. It IS obvious from.

the plant response tables that these herbi-
cides are not equivalent and cannot com-
pletely replace one another.

Considering silvex in particular. it seems
abundantly clear that this interesting herbi-
cide. which has been around for many years.
probably merits increased consideration not
only on the part of those now advising on its
present registered uses but also on the part
of researchers to ferret out new possibili-
ties. including combinations with other
herbicides. 

SUMMARY

KURON herbicide is a low volatile ester
formulation of silvex. having wide utility in
controlling broadleaf weeds and woody
plants. Over the years. the product has
gained acceptance for use mainly in aquatic
and turf areas. but published literature af?
firms a broad range of utility for this herbi-
cide and suggests possibilities for further
use development. Many plant species show

. excellent response and can be cont-rotted at

 'r :4 
. moderate space! of srlvex

in aquatic and turf weed control is noted.
Concerning the possible environmental

impact of silvex use. it is known that breakdown 'and loss in soil occur via microbiologi?
cal degradation. Silvex is degraded more
slowly than 24-0 but buildup in- soil does
not occur from annual increments. Used as
recommended on the label, KURON herbi-
cide presents no health hazard from inges-
tion or skin contact to humans, farm animals
or wildlife. A toxicological summary was
published in 1966"". 4-

SELECTED REFERENCES

I. Byrnes. W. R. el 1957. A test of 
2.4.5-TA and ATA on mixed oaks and asso-
ciated Species. Proceedings 11th Northeast-
ern Weed Control Conference:188-195.

2. Coulter. L. L. 1954. Silvex. a new seiective
herbicide. DOWN TO EARTH 9. No. 4:18-20.

3. Coulter. L. and J. W. Gibson. 1954. A prog-
ress report of field experiences silvex.
Proceedings 11th Annual North Central Weed
Control Conference;90-91.

4. COulter. L. L. and K. C. Barrons. 1956. KURON
for tur'l weed control. DOWN TO EARTH 12.
No. 2:24-25.

5. Dunhant, R. S. 1965. Herbicide manual for
noncropland weeds. USDA AgriCulture Hand-
book No 269.

TABLE 5. Comparative control oi certain herbaceous weeds with silver and

2.4.5-T. .  

 

OFTEN BETTER CONTROL WITH SILVEX
hedstraw (cleaversl ground in
hlacheyedsusan hemlock. poison
chickweeds henhit

evening primrose limpweed. spotted
liddleneck inewel

?rewood Irnotweed. Japanese

OFTEN BETTER CONTROL WITH 2.4.5-T

buckwheat. wild meyweed

colleeweed mesquite. velvet
daisy, oseye mexicanue
gooseloot mustards

indigo. curly pennyeress. field

ironweed. western peppemeed. field

Inesshelder
medic. hleck
millmeed. showy
black
spurges

spurry. corn

poorioe

purslane. common
sage, creeping
shepherdspurse

yellomochet

 

TABLE 6. Comparative control of certain woody species with silver: and 

 

OFTEN BETTER CONTROL WITH SILVEX

cums. cholla Er prickly peer

honeysuckle

maples

mulberry

oaks (live. Itlue. turkey. and others)
palmetto

persimmon Ten-s

OFTEN BETTER CONTROL WITH 

huclhrush

mesquite
chemise oak. white
hawthorne osage orange

tldhay

poisonoak

tedhud

rose. Cherokee
sagebrush. hinged
seltcedar
trumpettrine

yucca

sagebrush. hig
snowhrush
sweetgum

yerhesante

oowu to EARTH. Vol 23. No 4. Snqu run 31

'16519'

'Ih,

non-rad

,5 .lmk?ml? . h-?L

and. .LJ haul-IIer 

 



 

10.

12.

13.

15.

16.

17.

18.

Elwell. H. M. at al. 1954. Aerial applications of
herbimdes on oak in Oklahoma. Proceedings
11th Annual North Central Weed Control Con-
lerence:91-95.

Gates, Dillard H. 1972. Rangeland weed con-
trol. Oregon Weed Control Handbook: 1 26-127.
Gaylor. James and Alfred Houser. 1962. Three
years? results with silvex for aquatic plant
control in Oklahoma. DOWN TO EARTH 18.
No. 32-4. -

Hall. w. C. 1960. Control of various aquatic
weeds with silvex. Proceedings 14th North-
eastern Weed Control Conference: 476-482.
Hughes. E. E. 1967. Dormant boom-applied
spray treatments for control of saltcedar. Fle-
search Progress Report. Western Weed Con-
trol Conference; 23-24. 

. Klingman. D. L. and W. C. Shaw. 1971. Using

phenoxy herbicrdes effectively. USDA Far-
mers' Bulletin No. 2183.

Leonard. O. A. and W. A Harvey. 1965. Chemi-
cal control oi woody plants. California Agri?
cultural Experiment Station Bulletin 812.
Motooka. P. S. at at. 1967. Control of Hawaiian
jungle with aerially applied herbicides. DOWN
TO EARTH 23. No. 1:18-22.

. Mullison. W. F1. 1966. Some taxicological as-

pects of silvex. Proceedings 19th Annual
Meeting Southern Weed Conference: 420-435.
Newton. Michael. 1962. Control of bigleaf
213,121? with silvex DOWN TO EARTH 18. No.
1: 20-24.

Newton. Michael. 1972. Herbicides inforestry.
Oregon Weed Control Handbook.

Owens. C. B. 1956. Chemical eradication of
turkey oak. DOWN TO EARTH 11, No. 4:23-24.
Pierce. Madelene E. 1960. Progress report of
the effect of KURON upon the biota of Long
Pond. Dutchess County, Y.Y. Proceedings
14th Northeastern Weed Control Conference:
472-475.

32 DOWN TO EARTH. Vol 28. No 4. sin-nu 1973

TABLE 7. Comparative control ol certain herbaceous Weeds with silvex
and 2.4-0.

 

OFTEN BETTER CONTROL WITH SILVEX

badsmw groundcharry modic, black
honorcups groundiiry rnilkwaad. cot-no
hlachoyodsusao hamlock. poison ?no
chicltwuds hauhil putslana
doglonnal hots-wood 
liddlenaclt knapwecd. spotted sputry. comm
Iii-wood toauiral woodsorral 
Ilaabaoos lupirtas mow 
CD
OFTEN BETTER CONTROL WITH 2.4-0 (0
CD
hluowud. Tom Ito-l- pinaapplowud
ironwacd. wcstatn pootioa
knapwaci'. o'illusa punctmairirto
croton oluicatuaa sage, 
dcatltcarnas mdplantain smartwud
garlic. wild mustard: spudwall
iioid nllomodiat

 

TABLE 8. Comparative control 01 certain species with alive: and

 

2.4-0.

OFTEN BETTER CONTROL WITH SILVEX
alder persimmon. common
blackberry locust. black poisoniiry
cactus maplcs poisonoak
cherry. wild mosquito raspbarry
choltochorry mulberry rodhud
alt-o oaks saltcadar
hickory osaga onugo mo oI heaven

palmetto 

OFTEN BETTER CONTROL WITH 2.4?0 
buckoyo cottonwood snowbrush
buckorush maroanita sagebrush. big
chamisa rabbithrush loyon

. '16520

Program Your Aquatic Weed Control



DD
0 1
ill-'-


Needs
By -

CHARLES T. LICHY

-_Plam Science Research and Development
he Dow Chemical Company
Winter Park, Florida.

title of this?talk is PROGRAM YOUR AQUATIC
VEED CONTROL NEEDS. What does it'mean to program?
)0 we realize what the word program can imply? Webster
lc?ncs program as a "plan of future procedures." An apt
le?nition but one which does not completely describe all of
he side cliects of programming. Anodtcr detinition listed for
irogram is ?a doctrine, theory or system whose validity can

it: tested only in practical application." This, perhaps, is

?ven a better de?nition for us working in weed control. Basi-
?ally, however, the word programming means ?what are we
tying to accomplish" and ?how can we go about With the
ask at hand." One way to do this is to list what the problems
tight be in regard to the type of aquatic weed ?problem?
On have. "Problem" would include not only the species that
ire present, but which Species you would like to have present,
if perhaps what is the purpose of the weed control program,
to it drainage ditch, irrigation canal, farm pond, waterfowl
ciuge or what have you. Other points that need to be listed;
what are the rami?cations of your control program, is spray
lril't a problem, is it a water Shetltarca, how about ?sh popu-
ution, is the area used for swimming, irrigation or what
:lse'g One factor that musi he considered is that you obviously
:annot Use a tlill'erent material for each species present. There-
IUTC. you must use a material which has a broad enough
so that the primary weed populations will be
etnoved. Perhaps the next year, then, the material will have
0 be changed to pick up the. resistant weeds that Were not

killed by the ?rst year?s application. Another factor which
may be important is ecology. After you spray one weed
pOpulation, what changes can be expected in the population?
And, of course, last but not least, are the economics involved.
Of course, they are always important'but sometimes these are
relatively important, sometimes they are relatively unim-
portant when looking at the total program. in short, pro-
gramming your aquatic weed control needs actually becomes
very complicated when all factors of control are considered.

Let us examine some of these factors in detail so that we
can arrive at principles where these factors might be taken
into consideration in programming our control needs. Per-
haps the most obvious place to start is which weeds do we
have to control or which weeds are our problem. it is not
necessary to go into any discussion on identi?cation of the
various aquatic weeds since these have been discussed in
some detail before the society on several occasions. One
point, however, that must be considered is the considerable
variation in the time when the aquatic weeds are most suscep-
tible to chemical treatment. For some species like the Arrow-
arttm (I?t'lrmulm star.) the most susceptible period is during
the ?owering stage. For Other plants such as the rose mallows,
hibiscus species and cattails ('l'yp/m spp.) the time of_ greatest
susceptibility is during the late llowering anti the early stages
of seed production. Gianteutgrass can be killed most easily
during the period of maximum runner growth, which usually

18521-

AMDG

880188

riitc'nds about a month after the initial flowering period.
.Vitlt the phragmitt?s tlte most susceptible period is at the
in": of flowering or pollination, two to three weeks after llte

Cattails are most easily when lit the late
Ibw?ring or early fruiting stage. lit the south the tropical
:attail often flowers about two months after the narrow leaf
:attail. For this reason treatments sometimes have
required to control both species. Farther north, however,
he period between flowering of the two species is much
better. in Maryland, for example, tropical cattail flowers
thout one month narrow leaf cattail and one treatment
as given satisfactory control of both species. The best time
'or treatment of needlerush occurs during the flowering stage
ll?ld'lt'lSlS for three to four weeks. Regardless of all this,
towever, Spraying can and should be done over a wide range
if growth periods since satisfactory results can be obtained
frequently less chemical is required because of the
mallcr size of the plants. Whether the plant sites are ?ooded
)r non-flooded can also play an important role in determining
:ffective control from a herbicide.

Aquatic plant control is also complicated by the need to
:onsider the role of plants in different situations- Weeds are
lescribed as plants being out of place. Thus, plants that are
indesirable in one location may be bene?cial in another.
fo the waterfowl manager a weed is a plant that does not
)rovide enough food or cover to justify the Space it occupies.
:or example, buttonbush is responsible for 70% or more of
he coppice growth which is a problem in the marshes and
.wamps of the lower Mississippi valley area. Yet in Mary-
and, Virginia and the Carolinas buttonbush contributes to
raluable waterfowl habitat. Phragmites also can have a place
it waterfowl management. The root growth will protect a
na luring storms. Aisu, the furnishes excellent
over for both the-hunter and the hunted. For example, in
last years it has helped to make the Thousand Acre Marsh in
)elaware one of the best duck shooting areas along the
\tlantic Coast. But solid stands are undesirable since landing
areas are not available. 

Since water is intimately associated with aquatic weed
ontrol'many problems are unique to the water weed control
eld. Aquatic plants differ in physiology and habitat so sig-
i?cantly from terrestrial plants that one cannot con?dently
pply to aquatic plant control the information gained from
tpericnce solely with terrestrial plants. Water quality may
an extremely important factor. For example, work done
The Dow Chemical Company shows the effect of pH on
Jontail as shown itt Table 1. Similar results were also
btained with potamogeton and alligatorwecd. Table II shows
he effect of calcium as well as the effect of pH. Notice that
he presence of calcium in the water. particularly at a pH of
or above, improved kill of coontail. Water temperature
my not influence the reaction of a chemical to a significant
Cgrce but the aquatic may be in their most active
r0wth at higher temperatures and thus would be more


Contamination of water by herbicidal chemicals is of prime
nportance. Naturally, all due caution should be exercised
then using herbicides. lt is necessary to ttse those materials
thiclt are relatively non-toxic to humans and fish as well as
ossible desirable plant species along the edges of the aquatic
rein/his thus very important to control spray drift; spray
rift ?tis lit-int: lltat physical drift. which throat-.1!
Pressure spray equipment or otherwise negligent use _of
tray itt which particles of the spray material
love onto desirable areas such as vegetables, citrus trees,

home lawns, gardens, etc. Some of these problems catt be
avoided by ttsing well trained and well equipped spray crews.
Spray crews should be instructed or shown how to exercise
care in proper chemical application. Materials should be used
which have. a non-volatile nature. It is important to use low
pressure to provide largest sized droplets that are compatible
with the type of application and control necessary. The end
use of the water should be considered. That is, whether the
water is ttsed for swimming, irrigation, drinking, fishing;
whether it is a water shed area, and some attention must be
paid to this when selecting herbicides for weed control use.

Practical considerations also be included in your
choice of weed control materials. You obviously cannot use
a different - material for each species you may
?nd along a ditCh bank or canal. You therefore, use
a material which will control the primary species involved.
This may require over a period of time that two or more
herbicides be used if eradication is desired. It may even be
desirable to make combinations of herbicides so that all the
species are properly controlled. In this regard attention must
be paid to the ecology, that is, the changing population after
a herbicide application. in the case of waterfowl refuges,
where certain plants are desirable, this can be made to work
for you. Table shows the effect of two chemicals on the
ecology of two plots before and after chemical application.
Note that before application the plant composition is approxi-
mately the same, yet after application, the plant composition
is radically different.

A farm pond presents a somewhat different situation in

aquatic weed control since you have a substantially stable.

water environment as contrasted to a ditch or canal. This
means that chemicals applied to the water remain in contact
for a long period of time. This has advantagcs in that 
to make the chemical more effective in weed control but has
a disadvantage in that it continues the water pollution problem
for along period of time unless after a period of weed control
the pond can be flushed with fresh water. Ponds have an
additional advantage in that construction of pond edges that
drop off quite rapidly and fertilization which shades the
aquatic weed serves as a reasonable means of weed control
without chemicals. However, once weeds get into a pond,
then chemicals must be used to eliminate them. Weeds may
geriato the pond front seeding upstream or birds scattering
seeds or a number of other ways. 'Water weeds often grow
in water stored for irrigation. They clog outlet and inlet
pipes, spray nozzles and irrigation structures. They also
harbor mosquitoes and interfere with fishin". Many irrigation
reservoirs are shallow and have flat bottoms and are subject
to frequent draw-down and re?lling. In this case neither
deep edges nor fertilization control weeds in such impound-
ments because of the fluctuating water levels. in deeper
reservoirs or those in which draw-downs are infrequent,
fertilization and deep edges may be practical. However, in
irrigation ponds extreme care must be taken so that waters
which have been contaminated with chemicals are not used
for irrigating sensitive crops. Lakes have similar problems
and normally not being equipped with deep edges and not
being practical itt most'eascs to fertilize, the problems are
considerably ntag-niticd. 

Let?s discuss some of the chemicals that are available for
control taking into account 50tth Uf llIL? prin-
ciples and problems we have discussed above. I'm sure all
of you have seen recommendations out by ill-Whitler

and Lyle Weldon from lit. Lauderdale. One of the 

you will notice hardened in almost all of the aquatic wcetl

680198 Moo

16522

 

a

 

0




 it The Dow Chemical Company

nun of silves? as the propylene
11:; is not a new material. it has

. rides for aquatic weed C01!ltr,l_



Karon herbicide at 5 qts/acrc
the troublesome submerged
States. Experimental tests in
. of tlte country evaluated
on several aquatic
in 1958 and subsequent years

nuance information. Karon con-

in Table w.
can be used. Emergent

(an: Kuron at I gal./lOO gal. of
emerged foliage as a thorough
for the control of submerged
:ane by diluting Kuron at 1 gal.
- and directing this solution on or
pumps are also often used_to
 and pump it to the boom which
an over a wide swath. Uniform
.- desirable to allow a lethal con?
-me into immediate contact with
auctions of undiluted formulations

.1 on a limited basis with varying

completely safe to fish. Experi-
's in several ponds have caused
areas where high concentrations
?lications at concentrations of 5
ante ?sh kill in a limited number
can no ?sh injury in over 99%
Rapid incorporation of the spray
associated with rates tn excess
tc avoided where ?sh kill cannot
'4 Jersey found that applications
on oxygen content in treated
with Karen normally decompose
:n to twenty days are required
decomposition apparently does
of the water. This is a problem
vhich give a rapid kill of weeds.
"-trnents with Kuron showed no
entltic organisms. Applications

'1 populations of some plankton
?catment the populations were

plankton studied consisted of
?terions etTect on large aquatic
:rdes in treated areas. Proli?c

it soon after treating aquatic
fides. Algae growth does not
four weeks after treatment
trance of algae apparently is

the aquatic Weeds and thus

etc the water from decaying
toxicity and presents a low

'1 use. The Karon herbicide

ll species attd the following

'ltou'n in Table V. Front

that there should be no

to the handling and list:

:itle should he left where

it. Cattle. sltcep_ swine,

water containing 

period with no adverse

effects as determined by water consumption, body weight
gains and growth observations including gestation and par-
turition itt sheep. These data indicate that foliage or water
treated with Kuron herbicide as recommended should not
present a hazard from ingestion. Skin irritation tests con-
ducted upon rabbits have indicated that Karon herbicide
and its dilutions are irritating upon repeated prolonged
contact. in these studies there was no evidence of absorption
through the skin itt acutely toxic amounts. The observation
of reasonable care attd personal cleanliness practices should
be adequate to avoid skin dillicultics. Karon herbicide when
diluted to 1% with water or 1 gal. per 100 gal. caused no
primary irritation or allergenic responses when applied to
the skin of it'fty human volunteers using a repeated insult
technique. in addition, the product was patch tested on a
second group of ?fty huntan subjects at concentrations up
to 50% without cutaneous reaction. Even when applied
undiluted to the skin for two ltours it caused no skin response.
Thus it may be concluded with a high degree of assmance
that when used as directed, this product should cause no
skin irritation or skin sensitization responses.

Karon may remain in water for three to four months after
treatment where no dilution occurs. Accordingly it is sug-
gested that water treated with Kuron should not be used
for Spray purposes or for irrigation. 

Thus Kuron appears to present a possible solution to a
wide variety of aquatic weed problems. It has a wide spectrum
of activity_on both submerged and emerged weeds, is rela-
tively low in toxicity to ?sh and other aquatic animals and
it has been exhaustively tested throughout the years. The
scientists at Ft. Lauderdale and other places around the
United States have had Kuron in test. in the laboratory and
in the ?eld. The Corps of Engineers has put out a large scale.
?eld test for the control of alligatorweed. Since Kurott does
have a wide spectrum of activity it is very useful where
minted stands of aquatic weeds occur. For example, in areas
where alligatorweed and water hyacinth are intermingled,
Kuron offers a solution for control of both species. Many
chemicals such as and others will control water hyacinth
but will not appreciably atfeet the alligatorweed. Karon
offers economic and ef?cient control of both.

For the control of aquatic artd ditch bank grasses a material
generally recommended is dalapon or the Dow trademarked
product Dowpon or Radapen. This material, like Karon,
has been extensively ?eld tested in a wide variety of locations
and species. It is particularly useful itt drainage ditches since
grass and eattails can seriously reduce the flow of water.
Cattails are particularly troublesome. They interfere with the
proper utilization and maintenance of drainage and irrigation
canals by reducing the velocity and volume of flow, and by
causing deposition of silt and debris. Safety can also be an
important consideration. In weed clogged drainage ditches
water. is _held for a longer period of time after storms, itt-
creasing chance that small children may tumble in and
drown. reservoirs, farm ponds, marshes and lake margins
eattails waste large quantities of water, crowd out plants
which provide food for wildlife and often interfere with
?shing, boating and other uses of tltese bodies of water. An
excellent bulletin on this has been issued by the U. S. De-
of Agriculture, July 1963, entitled ?Studies on the
Control of Common Cattail in Drainage Channels and
Ditches," These studies indicated that Uuu'pon herbicith
was extreetzely elfeetive in controlling cattail. Additional
work has been reported in Michigan, Otegon and other states
which shows the etl'eetiveness of Dowpon itt Cl?-




a.
CD

.1

16523"

TABLE IV
Aquatic Weeds Controlled by Karen at 1.25, 2.5 and
5 Qts./acre foot.

tails. Ted llall ol the U. S. Fish and Wildlife Service reported
the results applying Dawpon by aircraft in the southeast. In
this test Dowpon was applied to control cattail, ntaideneane
Md giantcutgrass. All showed good response. In addition,

.VPOH is non-toxic to lish and wildlife species as well as
kcroniestic animals.

Brusli control can also be important in certain cases of
aquatic weeds, particularly in ponds and ditch banks. Brush
control has been done by or combinations
of both. All of you are laniiliar with these results. Recently
The Dow Chemical Company has discovered a ncw'brush
control material trademarked Tordon. Although this material

1.25 quarts/acre foot or less (05 ppm) 

White water lily 511p.)
Pickerclwecd (I?onrerlcria ('urdum L.)
2.5 quarts/acre foot or less (1.0 ppm)

Yellow water lily .-.. (Nap/im- rpp.)



 

Mud Plantain --Il 5M1.
Water inilfoil 
5 quarts/acre foot or less (2 ppm)

 

 

 

 

 

 

is not yet in the marketing stage for aquatic weed control, (Utrrculnrm 5pm.)?
excellen brus control results have been obtained on utility (Balsam ?'hmbe") 
rights??way. The material is also currently?undcr lest Coontatl dentersmn)o
throughout the United States with aquatic weed workers. Fanwon (01130"le ?3

Walerwecd -. (A nae/tarts mnudensrs) 

In summary?remember that a uatic weed control is . . 
Variable pondwced (Poramogcron drrersriolrus)

perhaps more complicated and has more rami?cations than
terrestrial weed control. Be sure to take into consideration

(Patamogcron umplt'lolius)

 

 

 

 

 

 

 

 

 

 

 

all the problems and possibilities of problems in controlling Najf?d -- (NON-Y 
aquatic weeds by chemicals. The chemical companies are Spll?c ?15h 
aware of your complicated problems and are constantly 
striving in research and development to offer better solutions watersmr?m? spp.)
to those problems. it is up to each of you to be aware not Soflt rush - I (JUNCHJ 
only of the problems of aquatic weed control but also the 
latest solutions to these problems. (Sparganmm spp.)
TABLE .
Effect of pH of Water on Silvex Effectiveness ??50 Value-'5

Silrex Kill I . - 
no rating Animat . F, 
5 0 0 0 Species - B?s/kg
7f 0 Rat 1070
0 Guinea pie .. 850
5.0 3.5 - 10 Rabbit 350
7.0 3.5 3 Mouse 2140
9.0 3.5 3 Chicks 2000
TABLE It Bibliography

Effect of Ca and pH on Response of Coontail

to Silvex a! 2 5 1. Bull, Edwin, "Results in Applying Dalopon by Aircraft to Pest Plants in

Waterfowl Area: of the Southeast," DOWN TO EARTH, Summer, l958.

Ca Kill 2. Day, B. E. and Stirezey. A. W., "Control of Phragrnites with Dolopon,"
pH rating DOWN TO EARTH, Spring 1956.
0 5  3. Goodrum, Phil 0. and Gray, Charles E., "Experimental Control of Giant
0 7 7 Cutgrass in Eastern Texas," DOWN TO EARTH, Winter I958.
0 9 2 l. Grigsby, B. H., Reimer, C. H. and Cutler, W. A., "Observations on the
 5  control of Cattail, Typho Spp., by Chemical Sprays," Quarterly Bulletin
250 7 9 of the Michigan Agricultural Experiment Station, Vol. 37 (3) pp 400-406,
February l955Heath, R. G. and Ruch. L. C., "Aerial Control of Cattoil With Rodopon,"
500 5. 10 DOWN TO EARTH, Winter 1957.
500 7 9 6. Pierce, M. E.. "The Effect of the Weedicide Karen on the Flora and
500 9 8 Fauna of Two Experimental Areas of Leno Pond, Duchess County, New
York," Proc. N.E.W.C.C. l2: 333-343, I958,
l" 7. Pierce. M. E.. "Progress Report of the Eflect of Karen Upon the Biota
Ptant Composition as Affected by Chemical Treatment of Long Pond, Duchess County, N. Proc. N.E.W.C.C. M: 472-375,
l960. 
not I Id": All" 8. Steenis, J. H.. "Aquatie,Weed Control," Proceedings Forestry symposium,
Before After Plot 2 August 30 and 3i. I960, The State University.
. . r' r? 9. Timmons, F. L. et al "Studies on the Control at Common Cottoil in
"9313?th  1% 95 207? Drainage Channels and Ditches," USDA Technical Bulletin l236, July
3% 3% 70% I963. 
)oct-z-o  939;, 2% IO. Weldon, L. W., Summary Review of Investinotions an Allinnloweed
- . and its Control," USDA, Corps ol Engineers, CK 33-60 September l9ti0.
Jill?s Trice '1 race 2% . . ..
. ll. Wiltse, M. 6.. "Annette Vegetation Control With Kmart. Presented

5% at l960 meeting at the Weed Society ol America.

18524 -

 



I

-

. 
I
. .
?unin.JIno.-.. . - J'rrgv cn-"I-h-.- sy??h-g' - - - . -
..nllb .. 0-.-- . . adve.ruinpr- 4:..- aunt-co? If-..4,125.-.. .. - .. I.-- - .Lf??Il?v 
'42 .- i

  

OFFERS A CONTROL FOR UNWANTED VEGETATION IN AND POHDS

HE DEFINITION-OF-THUMB says a weed is ?a plant the chemical must have a wide margin of safety to
out of place.? A weed can be out of place in a birds, animals, ?sh and humans. 3
garden, in a lawn, in a farm ?eld . . . or in a lake. The recent acceptance of Karon as an aquatic
Finding suitable methods of controlling unwanted weed control agent marks a big step forward in this
aquatic vegetation has been a goal of agricultural ?eld. Karon has been known for years as a brush
chemists, conservationists and water Sports enthu- killer, eSpecially effective on hard-to-kill species such
aiasts for years. as oak and maple. The new use registration has been
The nature of aquatic weed problems depends granted by government regulatory agencies on the
largely on one?s aquatic interests. Odors that ac- basis of performance coupled with substantial safety
company excessive aquatic weed infestations are in use. 
offensive to sightseers. Vegetation can jam boat The story of Truesdale Lake shows the possib'ili-
propellers, trip water skiers or ruin swimming spots. ties for Karon in this new ?eld.
Anglers know that excessive vegetation can deatroy Truesdale Lake is an attractive body of water in 
?sh spawning beds or unbalance fish populations. New York State. The lake is ringed with summer
And to industry, excess aquatic growth in cooling homes and was a popular watering spot until it
ponds can interfere with circulation ol'cooling water. became choked with weeds which virtually ruled
To qualify for aquatic use any herbicide chemical out swimming, boating and ?shing. Members of the
mufnmeet rigid specifications. First, it must do a Truesdale Lake Preperty Owners Association faced
got. Job of controlling aquatic plants. Secondly, decreasing fun and declining property values and .

.-

 

Ropnnted from Aprll I959 Dow Diamond:

.1652,

ol irueadala [pl-o in-New York before treatment
have veqel'olion. The lake has an area of 88
and ?future! are populated with summer ham".
1. pam several year! weed: had interfered
uaiingty with boating and ?itting

Treating the lake with Koran to remove vege?
totlan. A mixture of the chemical and water
we: applied as a coarse spray over the tur-
face of the lake. Karon formed a white emul-
sion in the water, persisting for about 20
minutes and eliminating the need for marker
buoys. Kuran is effective in controlling some of
the mall obnoxious aquatic needs when as
little as Iva part: per million are applied in
lake water. The treatment was made by Robert
Sheridan

knew that something had to be done. ?Something?
proved to be a big order. Various materials were
tried with discouraging results. When association
members heard of successful tests in New Jersey
using Karon to control aquatic weeds, they deter-
mined to try the method on Truesdale Lake.

Necessary arrangements were made with the City
and State of New York to make the application last
summer. Since the lake is a part of the watershed
serving New York City, special arrangements were
made to measure chemical concentrations in the
water.

The program was conducted under the general
supervision of Dr. E. C. Haney, professor of fisheries
biology and zoology, Cornell University= with assist-

 

 

It: .- - I
.. a-n?tau?'rt-ue-I-ance from Roy R. Younger - of the New Jersey
Department of Conservation. Robert Sheridan, 3
custom spray operator from Dover, New Jersey,
was engaged to make the treatment.

The entire operation came oFt? with no adverse
effects, save a minor kill of ?sh. This seems almost
inevitable in aquatic vegetation control work since
decaying vegetation ties up oxygen in the water.
However, Karon kills aquatic plants gradually, hold-
ing this problem to a minimum. At Truesdale Lake.
cottage owners were glad to overlook the slight ?sh
kill in favor of greatly improved ?shing conditions.
And this past summer, for the ?rst time in five
years, boating. swimming and ?shing were possible
and pleasant in July.

"my?1:2! .

 

'h a-i: 52-w-

"990168 Mo

Four weeks after application
thalalte had become IO clear
that the bottom could be seen
in many areas. Observers
watching the operation re-
ported a gradual kill of we-
tor weeds. in about a week
after application plants be-
gan turning down, with man

.67 . 
e. at? 2,1?
1 .r .
1-:  -. ?gov-?run-:-  Hi". I) f.   *Fr.Qf. zi't?Wu-hen ?d-?cuew . .
uni-ubweeds going to the bottom
Move-ks or so after appli-
- I: 
I cation
- - - -  1-5743??atHu?urb'. when far superior to the Ll)? are ad-
111111151rre1l). and high potency as an enzyun- inducer 
suggest that this could he of a casr'ade-type involving a series
of lnochrmical lesions. We have therefore investigated the
effect on the enzyme equipment of rats intoxicated with 
so as to define the targets of this biological damage.

Wistar rats (111,; and 1012) weighing were
given single dose of 111 1111:: of (?Ike in olive oil by intra-
peritoaeal injection: the controls (111.3 and 1119) received the
Solvent alone. Both treated rats and controls received a normal
and water ml 11111111111. After 111 days, blood samples were
taken by cardiac puncture and the levels of the follou'ing
serum enzymes were measured: glutamic oxaloacetic trans-
(15.0.1) glutaniie pyruyic transaminase 
ahlnlase lactic dehydrogenasu  

hydroxyhutyric dehydrogenase alkaline 

phatase arylesterase and cholinestcrase
(ChJ-i.) (111]. The reSults, analyzed statistically by Student's
t-test. are given in Table 1.

aide Effect of on serum enzyme levels

 

 

Enzyme SexS Controls Treated
o? 9 65.213; 4.11) 344431-4003?1
S.G.P.T.- o? 3233': 1.55 43.6-1i 3.117?
.1111.I a 9 27.433: 1.74 ?29.3ni 2.35e
s? 9 473.601; 31.73 716.33i76.S-1d
llb.DH.? 9 250.44 3; 23.111 339.921.11.021
.5 9 1.22 17.143h 1.20e
A 333.2oi19115 
<2 316.3011-130 213.60i (1.73?
Ch 2.13 moi own1
9 13.433: 1.61 3.2233 0.35cl

 

Expressed in Expressed in Bodansky units. ?3 For
results expressed in mg of phenyl acetate hydrolyzed in
1 by 1 ml of serum: for L?h.li., results expressed in mg of
hydrolyzed in 1 by 1 ml of serum. *1 Statis-
tically highly significant. Non-significant. 
I Separate figures for 0? and 9 given only when there are
statistically significant differences.

Table 2. Effects of (I) on blood serum components in rats

(6 and 9)

 

 

 

Component Unit Controls Treated
Glucose g/l 1.17 0.04 0.68 0.03?
Cholesterol. (:11 23710.1 2.65 3:015b
L'rca g/l 0.3-1 ?0.02 0.43i0.03?
Total lipids e/l 3.79:0.15 6.25 $0.43?
l'l?ult?ln till S7.?l  1.95"
lhhruhin Ineflouml 3101-11.} 23.57 14.59?
Sodium mliq/l ?1-10.11  2.41 130.25 3.31?
Potassium nil-311,11 4.57 3311.31) 5.42 :11; U.-H '3
Chloride mlitu?l 102.4 10.71 99.6 g; 1.5?

3 0.11111. Non-significant. 1? 0.112. ?1 Highly signif-
icnnt.

It is evident from Table 1 that ?dioxin produCes deep

perturbdliulh 111 aL'VL'fftl enzymatic systems in rats; the dis-
turhanCe may he :1 decrease (particularly remarkable in the
casu of choliui-sterase) or an increase (particularly considerable
in the case of 

Table 2. nhich I't-u-rds the effect of on the main components
of blond seruIIL shows a profound disturbance in the
lmmemtasis indicating, 11:111. that the live: is .1111: of
the main tutu-'1': 11f "dun-1111" intoxication. llistopatltohu-Ic
.t-vnlenu: of this damage to the and to other target 
(notably the thymus and heart) will be piese1:te1l in Separate
paper.

?111?:

Received January 17, 1972

fluu-ll111_ N. :1 111.: (LR. Acad. Sci. (Paris), Stir. 273,
7115 (1971). (2, m. 22.5 nail?: in the rat, 
in: liffeet: of Man and the Environment. l-leanngs

Knrze (lrigin' 'mitleilnngen

-qmwaewlg; 

?1 5111111151111? 11 

[if --1. 1"  
of the ('nntmiltI-e 1m l't11111nr1..1- (US. 51:11:1111), .\ Ifll 7 and 15?
19711. US. (inyt. l'rintinu (Mine, \?l'ashinetnn, ?funny?,
(1. 1911. Nature 220, 7112
(111115). L4) llnu-llui?, N. 111.: Cit. Acad. Sci. (l'aris),
S?r. 272, 5.: Amer.
J. Clin. Path. 28, 5(1 (1957). Shapira. (L, :1 Sem.
Paris 29, 11117 (1953). Wrohlewski, r! 111.: I?rnc.
Soc. lixper. lliol, Med. 90, 2111 (1953). ltosalski, S. 
Wolkinsnn, 11.: Nature 188, 11111 (1961)). (9) lindansky,
0.: J. Biol. Chem. 99, (1932). [10] Vincent. :1 121.:
Ann. Biol. Clin. 13. .1139 (191311).


Organs as Targets of ?Dioxin? (2.3.538- Iletra-
Intoxication

u-Hoi?, Pham-Huu Chanh, (Miss) Sesque,
M. C. Azum-Gelade, and G. Saint-Ruf

Centre National de la Recherche Scientifique. Paris, France

In previous papers we showed by means of enzyme
studies that the liver is a main target of intoxication by the
extraordinarily potent toxic and teratoeenic agent "dioxin"
The present paper
Supplies anatomo- and histopatholoeic evidence of damage to
this organ, and to other organs which we have also fOund
to be profoundly affected within the short duration (10 days)
of our experiment.

Wistar rats (10.5 and 109) weighing 2101-20g were given
a single dose (10 mg/kg) of by intraperitoneal injection in
olive oil; the controls (105? and recaived the solvent Only.
All' the animals received a normal diet and water ad fibilum.
The treated animals lost considerable weight as from the

rens: 732'

?5511' 

- . 
 use?. sate-?11 

A. . 5.. I 
1-11- "5"1?0 n't'

Fig. 1 :1 and liffect of "dioxin" intoxication on liver and
thy1nt.s. a) llepatuct'te deterioration: stmtusis, 
voluminous plasmodial forum (x 2511). b) inverted picture .of
thym?nistjuctnre x1115)

 

january 25. 1972.

 



i

..
d.
.

.t



16521-




. 

sit-:19the i-spt-rnm-nt, and luth day thev wriehcil
hiss Ihnn .il the sl..rt, [he controls had gained
f-tI-IHg. examination shunt-d a highly signif-
icant increase [Student's l-ti'st) in the liaematncrit (controls:
treated: 2 37), sienilicant increase in the
leukocyte (?nunl 5912;: treated: 73151-45?
uith a decrease at (controls: 79% d: 1.04:
treated: 3 II) and augmentation nl pulynuclcar neutro-
phils treated: Hawever,
no significant ellt?ct was made on the hone marrow within
this l-rit?l Pt'rinil,

All the treated animals had lie-pan: lesions, the females being
more severrly allected. The lubules were reshaped by im-
portant centrilu-liular stasis the llenincke trahe-
rulae (pronunnCt-d anisukarvosm, frequent hinucleation, cyto-
plasm loaded vacuoles and hyahne inclu-
sions]. and a focal hyperplasia of the l\'npller Cells: along-
side the lainucleatetl voluminous plaslnodes were
often present. These ?ndings were already prominent by the
6th (Fig. la).

Apart lrum these he Iatie lesions, which Confirm the biochem-
ical findings rrpt-rtei I. no other oreans manifested con-
siderable im'u-leement. The modifications sustained ll): the
ranged from ditluscd P}'kn05l? ol the thymocytes to
an inverted picture of the structure (migration of thymocytes
towards the medulla) or total involution; these lesions wure
manifest as early as the 5th dae (Fig. I This thymus involve-
ment explains the particularly pronounced toxicity of in
young aniniaL-i. The third ore-an to undergo considerable change
was the hrart, the animals presenting a valt-ulitis with erosmn

    
   

Buchbesprechungen

Buchlnesprech-I-ieen I75

ol the endneardinni on the ventricular l.u:i- nl thi- valves,
lilirinniil and oedema nl thi- ?huh ?t'fL'
thickened liy lilirnlilastn: prulileratiun. Focal li-smns lncatud
deep in the were also ?list-reed, with a dis-
appearance of myolilirillae and interstitial in-
liltratinn. the same time, ultra-arterial lilirinu-leukucyti-
thrombi, periarterinlar lilirohlastic reactions, and loci of
alveolitis were seen in the lungs.

These target organs sullered qualitatively the same damage
when a considerably smaller dose. ting/kg, at was
administered.

The pathological deterioration observed in the liver and heart,
and, to a lesser degree, the lungs, of rats, is Similar to that
reported by Allen and Carstens in an experiment with
monkeys on a long?term diet which included a toxic lat con-
taminated with a "chick edema factor" was later found
to contain chloro derivatives ol including (1)
not included in their study;

vain-s

"Ne thank Dr. s. s. Epstein, E's helpful dis- 


cussions and advice.



Received january 13. 1972

 

N. P.. at CR. Acad. Sci Sf-r 272,
1447 (1971). Buu-lloi, N. 13., c: .25.: Naturwissenschaltcn
59, 173 (1972). Allen, J. L. Amer. 
Vet. Res. 28. 1513 (won.- Lab. ant'SL 15, 970 (1900). 
Higginbotham, G. lt., .11.: Nature 221. 702 (1903);
Cantrell, j. 5.. :1 51.: Acta 25 B. (1969).

 

.-

Otto Hahn lEine Bildd-Jkumentation. Von Ernst Berninger.
Manchen: Heinz Moos 1969. ?165.. I37 Abb.. 

Das Each is: ein dokumentarischer lien'cht ?ber einen auBerge-
wohnliehen Jlensehcn und sehr erloiereichen Forscher. Die
bebilderte Biographie ist in sich rollstandig und 
es auch zu beldazen ist. daB historische, wissenschaltlieh
interessante Doitumente und ein Teil des Schriltwechsels ini
Arbeitszimmcr Otto llahns durch liriegseinwirkung ver-
nichtet ?'orden sind. Hahns Charakter, scin Charme. seine
Klugkeit \\'eisheit sprechen aus dent Buch, wie wir,
seine Schiller. ihn kannten. Ein \X'issenschaltler hohen Ranges,
54':th im chea and seiner Umwelt Stehcnd. kontakttreudig
uod heitcr cr uns als cine die in
ihrer .\rt anneal-ti: uar unit lileilnen wird. liin interessantes
Buch uber einen auch fiber cine
verganecne unil rielseitige :Lpoche der hohen Zeit der 
schalt in W. Seelmann-Eggchert (liarlsruhe)

elektro-
Konstanten und nichtlineare 
s'on iiristatlen. \'nn lt.l$echmann.

It. S. llearmnn und 5. K. linrtz Nelle
Serie. lid. 2. llrsu. und A.M.
llellwegi-l. Berlin-lleulellierg-New York: Springer 196?).
232 5.. wt. Abb., 37.4?.

Nor 3 jahre nach llt'm des 1. liandesder Gruch 
..Kristall- und (Ni-m- Berle) hat: sich ein
lirganZungshand als nutm'ndlt: erwit's?en. lir Neillie-
sliinnuinuen und lireamnneen zu den elastischt'n. piezo-
llt'll. Ill?lil eli-ktrimplischi-n 
Nlull' zn (la-ren 'lt-iulu'rnlur- unil Die
LiIi-ratur ht liis etua Inns erlatlt. lh-r 
Han-l Ist auth-rilem erin-itert tiller die
dritti-r (lrilnune, ulier die 
uptistlu-r in liristallt-Il (unit-r 
einzelner sI-hr eeuanrr Messengen t-un llrechuiiesinilin-s) und
hlx'r Hun-ztanten zwt-iler Urdnung. Die
liapilrl werden gi-niacht durch klar alIeelaUte
liinluhruneen. die aus der ersten Ausgabu


sind. Der Zusammenhang zwischen
den kristallogranhischen Eigenschalten wird (lurch \?iellache
Querveru'eise hertz-?stein. Ein eeineinsarnes. nach ISruttolormeln
alphabetisch geordnetes Sumtanzverzeichnis und ver-
bintiet den inhalt belrler Bande. Leider sind hierin keine Hin-
weise and die der nachge .wesenen Daren enthalten. lieidc
Bands ge'noreu zusammen und :ollten in keinem Institut lehlen.
das sich nut oder Kristallographic 
sei es grunrllagema??ig, sei es hinsichtlich der Entwicklung ncuer
Bauageregate. Cerade die neu zugergten lx'apitel
sind in diesem Articitsbereich von bcsonderer 

I A. Neuhaus (Bonn)

Tlteorie unit praktisehe Anwendunp, von 
nern. Analyse ir'. Ciienue, lid. 9). \'on
F. Unilaud. Jansscu. D. 'ihlerie und 0. Frankfurt]
Main: Altademische s'erlagsgeseliscialt 1971. 76o 5., ISU.

This treatise has 14-)7 rciL?a'Cl'lCCS and offers an extensive
discussion of complexes. mainly with inultiilentale organic
ligands, in ernviinetric, volumetric, photometric and llnoro-
metric quantitative analysis of virtually all elements. The
theoretical introduction is rather weak and rum-5 on the ar.
chaic hybridization theory; thus. on page 13, the possibility
is left open that square-planar (this redundant worn ought to
be "quadratic") copper (ll) complexes contain one 
tron. On the other hand, the general description of what
ligating atoms are pretercntially hound to a given central
atom is quite instructive. The text contains detailed prescrip-
tions for a large number ol speculic methods, and highly useful
tables. The Sources of errors and interference are carelully
discussed. This book. a tremendous amount
of critical study of the literature, can he recommended to
everybody interested in inorganic analytical A
specilic aspect is the discussion of derivatives.

C. K. jnrgensen (Gent)

Colour Chemistry. By R. L. M. Allen. London: Nelson 1971.
330 5., f. 

Uneeachtet der lawinenartit: zunehmenden liedcutune der
bleiben die technischen l-?arbstolle ein wichtiges.

memo lill?i??

16528

-.-- cue.-. sue-w

ivl. F. Hoiick. J. E. Frommer. S. C. McNe' .
M. Richtand. J. W. Henley.J. T. Potts. ..
chem. Biophys. Res. Commun. 76. IO U977).

2. Holtck. N. M. Richtand. 5. C. cNeiU. S.

. Holick. J. E. Frommer. J. W.
Potts. Jr.. Biochemistry 18. 1003 979).

J. M. F. Holick. J. A. Maanughli
J. T. Potts. Jr.. R. R. Anderson?. I. H. Blank. J.
A. Parrish. P. Elias. Science 10. 203 (I980).

4. T. Okano. K. Mizuno. . Matsuyama. N.
Nobuhara. T. Kobayashi. Reel. Trev. Chim.
Pays-Bar. 98. 253 (I979).

5.. J. W. Melski. L. Tanen urn. J. A. Parrish. 
B. Fitzpatrick. H. L. ich..i. invest. Derma-
tol. 68. 328 (1977). criteria for patients
with skin types I. and IV. generally of
European extraction are based on the history of
the usual reaction I the that hour of full sun ex-
posure in early su met. Skin type generally
includes patients of Asiatic. Anterican Indian.
Mexican. and crto Ricatt extraction. Skin

  
 

    

type V) gene ly includes patients of African
extraction."
6. l. H. Blank. . D. Griesemer. E. GouldJ. in-

rest. Denna oi. 29. 292 (I957).

 
   

'l R. Belsey. . B. Clark. M. Bernat.J. Glovvacki.
M. F. DeLuca. J. Potts Jr
Am .57.50ll97d).

8. D. cter. D. V. Kimbcr. l-I. 
J. Pit rial. 200. l263 Male weanling
Hol an rats were fed a diet de?cient in vita?
min but containing 0.47 percent calcium and

0.3 percent phosphonts for 4 weeks.SIM refers -

. M..B. Clark.?

to the ratio at "La on me inside ut
(scrosal side) to the incubation me um on the
outside of the gut sac lmucosal dide). There
were six animals in each group: values are
reported as the mean standar error.

9. W. F. Loomis.$cienre (I967).

F. Daniels. Jr.. P. W. Post. . E. Johnson. in
Pigmentatian: its Genesis a Biological Con-
trol. V. Riley. Ed. (Appleton-Century-Crofts.
New York. I972). p. 13. 

H. Al the time that Loomis proposed this theory. it
was not known that during exposure to sunlight.
vitamin D, is not directly formed in the skin ll-
4) and that vitamin DglmuSl undergo a series of
regulated metabolic hydroxylations to 1.25-
before it is biologically ae~
live (12).

l2. E. Havinga. Erpe entia 29. ?81 (1973).

I3. M. F. Holick and?J. T. Potts. Jr.. in Harrison's
Principles of Infernal Medicine. K. J. Issel-
bacher. R. D. Adams. E. Btaunwald. R. G.
Petersdorf. J. . Wilson. Eds. (McGtaW-Hill.

New York. I 80).
l4. 5. A. Levi ohn. Am. J. Dis. Child. 34. 955
(1927).

IS. Internatio al Commission on Illumination. CIE
Publicari '11 Na. 1972 (Paris. I972).

16. We th R. R. Anderson. J. A. Parrish. and J.
T. Pot . Jr.. for helpful advice and assIStance.
and 3. Fred. J. S. Adams. and S. A. Holick
for ttorial assistance. Supported in art by a
NA A grant (NAS9-15205) and NI grants
(A 25395-0] and 

8 July I980: revised I OCtober I980

2,4,S-Trichlorophenoxyacetic Acid Causes Behavioral
Effects in Chickens at Environmentally Relevant Doses

Abstract. Administration of the herbicide 2.4 .5-trich10rophenoxyacetic acid to in-
cubating chicken eggs alters behavior after hatching. Single doses, with no morpho-
logical e?'ects. retard learning (lowest dose, 7 milligrams per kilogram of body
weight) and increase general acrivity (27 milligrams per kilogram) and jumping (13
milligrams per kilogram). Day 15 ofincubation is the most susceptible srage ofdevel-

opment.

As far as we know. there has been on-
ly one earlier study reporting tcratologi-
cal effects of 2.4.S?I?chlorophenoxyace-
tic acid on behavior (1). Admin-
istration of. (100 mg/kg) to-
pregnant rats produced no morphologi-
cal del'ormities in the but it did
cause them to have learning de?cits and
to exhibit increased activity in the open
?eld. We have shown that even lower
doses can produce behavioral abnormal-
ities in the chicken. A single dose of
2.4 .5-T [Sigmaz containing 0.03 part per
million of 2.3.7.8-tetrachlorodibenzo-p-
dioxin (TCDD). according to the Austra-
lian GovernmentAnalytical Laboratory]
was administered to the chicken egg on
either day 8 or day 15 of incubation (2).
A third group received 2.4.5-T by intra-
peritoneal injection on day 2 after hatch-
ing. The was suspended in a 0.5
percent solution of gum tragacanth made
with Sterilized. distilled water. Control
animals (matched for each Separate
hatching) received the vehicle alone.

?Doses were calculated according to the

weight of the chicken or eg at the time
of injection and ranged from 7 to 53 mg/
kg for eggs and 75 to 225 mg/kg for
chicks after hatching.

SCIENCE. VOL. 21!. 6 FEBRUARY l98l

'The median lethal dose on day
2 after-hatching was 200 mg?kg. and on
day 15 of incubation it was 53 mg-?kg.
There was 70 percent hatching in eggs
given on day 15 of incubation at
doses up to 27 mg/kg. Some 5 to 10 per-
cent of the hatched chicks, irrespective
of dose. showed abnormal leg develop-
ment: they either dragged one leg or held
it off the ground. Approximately 5 per-
cent of the treated chicks showed depig-
mentation of feathers and dovm. Both
of these effects have been reported (3).
No di??ercnces in body weight gain were
observed in chicks receiving 2.4 .5-T dur-
ing incubation or at doses less than 150
on day 2 after hatching.

The chicks without apparent morpho-
logical dcforrnities (after doses shown in
Fig. I) were given behavioral tests in
week 2 after hatching. The tester was un-
aware of the treatment that each chick
had received. There were 8 to 20 chicks
in each group tcSLed. We measured the
activity in an unfamiliar environment on
day 7 of life by placing the chick in a
gray-walled box (walls). Cumulative activity for a 3-
minute interval (?general activity") was
scored by an Animex aetivity meter. and

oorssorssHome-0593500500 Copyright 193) 

gut 54C

number ofjumps made was counted

by eye. We scored ambulat' tubs-,5 
groups by dividing the ?oq?o?D?-?ox
into quadrants and counting the number
of times the chick crossed into a new
quadrant. We tested visual discrimina-
tion learning on day 9 oflife by using the
?pebble ?oor" test (4), which consists of
a search for grains of food scattered over
a background of small pebbles adhering
to the ?oor. Within a 60-peck trial. a

measure of visual learning is generated 3:
by recording the number of incorrect re- 2
Sponses (peeks at pebbles) within con- 0

secutive 20-peck intervals. 

- U1
General activity, jumping, and visual 
learning rate were altered by 2,4,5-Tu

treatment (Fig. I). Since the same doses
were administered incubation, these data could be exam-
ined by a two-way analysis of variance
(that is, dose, age. and their interaction).
A one-way analysis of variance was used
for the data from day 2 after hatching.
General activity scores conformed to a
parametric distribution, but jumping and
learning scores required logarithmic
transformation.

Jumping behavior proved to be the
most sensitive parameter a?'ected by
For those treated on days 8 and
15 of incubation, jumping showed both a
signi?cant dose e??ect (F 2.78: d.f. 
3, 90;? .05) and signi?cant interaction
between age and dose (F 3.33;
d.f. 3, 90: .05). Comparisons of
the groups given the two highest doses
with their control groups (t-test) showed
a signi?cant increase in jumping in the
groups treated with 13 (P .05)
and 27 (P .01) on day 15 of in-
cubation. but there were no signi?cant
differences in those treated on day 8 (see
Fig. 1). There was also a signi?cant dose
effect for jumping in the groups treated
on day 2 after hatching (F 4.06;
d.f. 2, 20: .05), and a Nest of
data for the control group and the group
receiving a dose of 150 revealed a
signi?cant increase in jumping after
treatment (P .025), this dose
being tenfold higher than the lowest dose
found to be e?'ective on day 15 of in-
cubation.

General activity, which includes mea-
sures ofjumping, ambulation. and other
movements. shows a similar tendency to
increase after treatment on day
15 of incubation and day 2 after hatching
(Fig. 1). Analyses of variance just failed
to reveal signi?cant effects. but of
data for the control groups and those
groups that had received the highest
doses at each age revealed a signi?cant
elevation of general activity in the
groups treated with 27 mg/kg on day 15

18539



.3



-. museum. \r .uu and those recetv-'

ing i150 mg/kg on day 2 after hatching
(P .025). There were no signi?cant ef-
fects after treatment on day 8.
'A'mbulation. the other component of
- general activity that we measured. did
not show any dependence on dose and

10 signi?cant di?'erences were found as a?

result of examination of the data by t-
tests,? even though there was a tendency
for ambulatibn to increase with increas:
ing close after treatment on day 15 of in-
cubation and day 2 after hatching (Fig.
1). Thus. as found in rats, has
long-lasting e?'ects on' activity, but in
chicks the most sensitive parameter con-
tributing to. the increased activity is
jumping rather than ambulation. which
may indicate increased fear responsive~
ness to novelty.

There was a signi?cant dose effect for
visual discrimination learning in those
groups treated on days 8 and 15 of in-
cubation (F 4.19; d.f. 3, 86', .005
.01) 'but not for those treated
on day 2 after hatching. Comparison of
treated animals with their controls' at
each age revealed a signi?cant slowing of
learning in those treated on day 15 of in-
cubation with 27 mgi?kg (P .001) and 7
(.01 .025). but no signi?-



General activity

Day 
Incubation

it


200 Day 15
incubation

?00

?iit?ill-?Wai?Day?!
alter hatching

,l

100

 

Dose (mg/k0

        
 

cant e??ects were present after treatment

at the other two ages.

Day )5 of incubation appears to be a
stage of maximum sensitivity to the be-
havioral effects of Neuronal cell
division is at a peak on day 8 of in-
cubation and is largely completed by day

15 when synaptic proliferation is occur--

ring (5). Synaptic proliferation is com-
pleted before day 2 after hatching (4).
The brain therefore seems to be most
sensitive to 2.4.5-T at a time of maxi?
mum synaptic formation. The fact that
caused adverse effects at lower
doses than reported previously may in-
dicate an extreme susceptibility of the
developing nervous system and hence
behavior. This is consistent with one re-
port of the possible adverse e?'ects of
in the human Species (6).

The extent to which our results are
due to or to very small amounts
of its highly toxic contaminant. TCDD,
remains unresolved. However, it is im-
portant to remember that in this study
we used of greater purity. partic-
ularly with respect to TCDD content,
than that available commercially and
sprayed in the environment. Based on
the concentration recommended in Aus-
tralia for Spraying blackberries (7).

Ambulalion Visual learning

-

nitrate: ?t

       

{mittwhich is 2.8 liter/ha of commercial 80
percent 2.4.5-T (weight to volume). m1
would receive 224 mg of 2.4.5-T. Since
the surface area of an egg is approxi-
mately 50 cm?. the dose of 2.4.5-T re-
ceived by one egg after spraying would
be 1.1 mg or 22 mg/kg, which is greater
than the lowest dose found to be behav-
iorally teratogenic. As 2.4.5-T has been
found to pass through the egg shell (8). it
may therefore present a real risk to bird
species. It is questionable whether there
is a dose below which Can be said
to have no effect since some individuals
appear to be highly susceptible. At all
the doses we used. morphological defor-
mations were seen in some of the treated
chicks, and. as the raw data for jumping
in Fig. 2 illustrate. some individuals with
no morphological deformations were
highly susceptible to the behavioral ef-
fects of 2,4 even at the lowest dose
(7 mg/kg) that we administered.

There is considerable Species variation
in sensitivity to the toxic effects of2.4.5-
T. At present one of the best indicators
of interspecies toxicity is the rate at
which 2.4.5-T is metabolized and ex-
creted (9). Based on such measures.
chicks and rats appear to be of roughly
similar sensitivity whereas humans have

 

52 a I
mom
day 15 hwbation .
0-1 00-0.. 000.04 00.. I
0 7 13 27

Dose (mg/kg)

Fig. (left). Plots of means and standard er-
rors for four behavioral measures after the ad-
ministration of 2.4.5-T at the doses indicated
On day or 15 of incubation or day 2 after
hatching. Behavior has been tested in week 2
ct" life. The ordinate scales are as follows; for
general activity. counts on the electronic ac-

tivity meter in the total 3 minutes of scoring: for jumping. the number of jumps in 3 minutes: for ambulaticn. the number of times :he chick

c" "-cd with- both feet into a new quadrant of the box: and for visual learning. the number of errors (peeks at pebbles) in the last 20 pecks of the
.e-?oor learning task. In the latter test. a higher en'or score indicates slewer teaming. Th: asterisks indicate signi?cant di?'erences from the
control groups as determined by .05: .0): and .DOI.
obtained during 3 minutes in a novel environment aftertreatment with 2.4.5-T on day )5 of incubation. Ea..h point represents the score for a single 

individuaL The fact that some individuals are highly SUSCeptible to the behavioral teratological e?ects of 2.4.5-T is demonstrated by the  
that. even after treatment with the lowest dose (7 some individuals show a marked Increase in Jumping.

59-1

Fig. 2 (right). The raw data forjumping scores

SCIENCE. v01; 

.l

00

1

1

SESEZIZ

 at mm ?a threetotd greater sensitivity
Our ?ndings therefore imply a risk

to the human Species.
C. A. Smoensou
L. 1. ROGERS
Pharmacology Department,
Monash University,
Clayton. 3168, Victoria, Australia

I References and Notes

1. P.-O. Sjod'en and U. Soderberg. Pin-rial. Psy-
chol. 3. 175 (1972): in Chlorinated Phenoxy
Acids and Their Dioxins. C. Ramel. Ed. (Swed-
ish Natural Science Research Council. Stock-
holm. 1978). p. 149.

2. J. McLaughlin et al. . Toxicol. Appl. Pharmacol.
5. 760 (I963). 

3. H. Lutz and Y. Lutz-Ostertag. Adv. Exp. Med.
Biol. 27. 127 (1972); J. F. Dunachie and W.

w. Helcher. Nature (Landon) 2?15.  avail. 

4. L. J. Rogers. H. D. Drennen. R. F. Mark. Bram
Res. 79. ?974).

.5. B. M. Freeman and M. A. Vince. Development
of the Avian Embryo (Chapman and Hall. Lon-
don. l974): J. Sedlaeek. in Advances in 
biology. G. Newton and A. Riesen. Eds. (Wiley,
New York. I972). vol. I. pp. l29-l70.

6. 8. Field and C. Kerr. Lancet 1979?1. 
(I979).

Vermin and Noxious Weed: Destruction Board.
Keith Turnbull Research Institute Bulletin 35
(Department of Crew-n Lands and Survey. Vic.
toria. I977).

8. N. Gym-Hansen and S. V. Dalgaard-Mikkelsen.
Acta Pharmacol. Toxicol. 35. 300 (1974).

9. F. J. Koschier and M. Acara. J. Pharmacol.
Exp. Titer. 208. 287 (I978).

10. W. N. Piper. J. Q. Rose. M. L. Leng. P. Geh-
rinz. Toxicol. Appl. Pharmacol. 26. 339 
K. Erne. Acta Vet. Scand. 240 (1966).

Neurosecretory Granules: Evidence for an A ing Process

Within?the Neurohypophysis

 

Abstract. When cysteine labeled with sul?tr-35 is in eCted into the third ventricle of

the rat brain, it is ?rst incorporated into only one

 

secretary granules that can be isolated on an isost otic gradient. The second popu-
lation of granules is labeled much later. Stimulati of hormone release fr0m isolat-
ed labeled neural lobes and subsequent isolatio of neuroseCretory granules at dif-

ferent times after the injection of labeled cy

 

decreases in only one population of granules. One of the fractions of the gradient
represents the granules found near the relea site; the second population is prob-

ably located deeper in the nerve endings 
rophysins are found in both populations.

   

one. Thus it appears that neurosecremry armies undergo an aging process and that
isosmotic gradients can separate the ag granules from those newly formed.

Neurosecretory granules (NSC) in 
hyporhalamo?neurohypOphysial tract re

  
     
 

20.000 daltons), which pre umably is
packaged into granules. Th contents of
the granules mature durin transport to-
ward the neurosecretory nerve termi-
nals. When the granules ach the neural
lobe. almost all the pr ursor has been
converted into proteins of 12.000 2
2.000 daltons, neurophysins, and pep-
tides that may include the hormones
oxytocin and facopressin (2).

The neurosecretory axons in the
neurohypophysis can be subdivided into
three compartments, all of which contain
NSG L3): undilated axons COntaining few
NSG. nerve endings characterized by
the presence of microvesicles. and nerve
swellings containing secretory granules
but few if any microvesicles. We pre-
?Wiously 'showed that the mean diameter
Jf NSC: in the endings differs from that of
N86 in the swellings (4). Furthermore.
we showed that fractionation of N50
from the neural lobe of rats on an isos-

SCIENCE. VOL. Ill. 6 FEBRUARY [98]

the nerve endings. whereas the mean di-
ameter of the osmotically sensniv NSC.
resembles that of the granules in the
swellings.

In this report we present evi
the osmoticale insensitive gr
newly formed NSG (NF-NS that have
recently an'ived at the neu lobe and
that the osmotically sensit' NSG are
probably derived from th former. Fur-
thermore. we show that NF-NSG are
located to the nerve en ings. whereas
most of the aged granul (A-NSG) can-

 
  
 
 
 

  

 

 

we ?link (1 Bentley and cuacn for not immediately releas their contents
valuable advice. and are thus likely to be found in the
23 July 1980; revised 29 September 1980 swellings. Protein 5pc ms of a molecular
weight louver than th of neurophysins
can be demonstrated the A-NSG. This
suggests possibl% postmaturational
cleavage of intragr ular proteins.
Albino (Wistar) Inale rats Weighing 250
to 300 were one hetized with ketamine
chlorohydrate (1 mg per 100 of body
weight) and giv intracisternal injec-
the two populations of neuro- tions of [?S]cys cine that was obtained
from the suppli (New England Nucle-
ar, 500 Ci/mmgle) or by reduction of
[3?S)cy51ine (47 Cilmtuole) with 10 mM
teine shows that the radioacn'vity dithiomreitol i 0.1M phosphate bu?'er
(pH 7.0). Eac rat was injected with 10
,u.l of the ?nal olution 1.50 ,uCi of radio?
in the nerve sit-ellings. Whereas nEtt~- active cystein The rats were decapi-
naller proteins can only be detecwd in tated at differ 1 times after the injection
and their neu lobes were dissected out
.in less than minute. Immediately after
1 their isolatidn. ten of the radioactive
motic gradient yields two populations of lobes were/ homogenized in 1.0 ml of
N80. each of which contains ncumphy- 0.3M sucrpse bu?'ercd atpH 6.8 with 10
sins, oxytocin, and One [Tl/9'1 Hep-s, and mixed with 20 non-
population has an isopycnic density of radioacu neural lobes homogenized
1.13 glcm? and is insensitive to osmotic under 1 same conditions. The N86
changes in the surrounding medium, were is ated as described by Nordmann
whereas the other sediments ata density et al. The gradient fraCtions were
of 1.1] g/cm3 and is much affected by the collect with a Buchler device, and a
osmotic pressure of the surrounding rne- portio of each fraction was kept for a
diutn. Furthermore, the mean diameter detcmtination of density. Scintillation
of osmotically insensitive NSG after ?uid was added to the fractions and their
ation is very similar to that ofthe NSG in total radioactivity was measured with a
Fig. 1. Time course
of the appearance of ?5
[35S]cysteine in NSG 
fractions isolated on 2
a sucrose-mentamide :3
isosmotic gradient at 
different times after 3
the injection of the 3;
isompe. The radioac- 
tivit_y in NF-NSG (0) 
and A-NSG (O) was 
measured and the re- 1;
sults are given as the .
percentage of the total j,
radioactivity found in . . .
both fracu'nns_ Hour: Day:

ooseaovstsuozoeos-tssoosoo Copyright 

9898212 tint]

Time alter intecuon ol 3 

.595 



 

"gm-L. - 

 

New Scientist 12 February 1981

MK. 

 

 

 

 

1

2



l?

Herbicide is ?real-risk? to birds and people C:

Human beings are at risk from the con-
troversial herbicide according to
two Australian scientists writing in the
current issue of Science (vol 211, 593).
The Scientists are seriously worn'ed by
the results of a recent series of experi-
ments in which they found that eggs
exposed even to small doses of the
chemical often developed into chicks
with impaired learning abilities. As
expected, the chemical killed some'of the
embryos outright. Humans are approxi-

mately three times more sensitive to

than chicks.

Previous research on 
trichlorophenoxy acetic acid) had shown
that rates in the uterus given fairly big
doses?100 milligrams of the chemical
for every kilogram of the mother?s body
weight?were more active and learnt
more slowly than other rats. These doses
are higher than those that are likely to
occur when farmers spray crops or
pastures with the herbicide. 50 C. A.
Sanderson and J. Rogers of Monash
University in Australia set out to dis?
cover what happens to chicks given
smaller doses before they hatch.

The scientists gave single doses of
(containing 0-03 parts per million
of the highly taxic contaminant 2.3.7.8-
tetradilorodibenzo-para-dio?n) to chick
eggs at two points during incubation:
the 8th or the 15th days. The doses
ranged from 7 to 53 mg/kg body weight.

These (105% were high enough to kill
some of the embryos: for example, half
the chicks given 53 mg/kg on day 15 of
incubation died. The herbicide caused

birth defects?such as depigmentation of 

the feathers or abnormal development of

the legs?in another 10 per cent. Sander-
son and Rogers tested the chicks that
were not obviously damaged to discover
any effects the herbicide may have had
on their behaviour. They found that
a?'ected three measures of the
chicks? behaviour: their general activity
in an unfamiliar environment; the num-
ber of times they jump; and how well
they learnt to discriminate visually
between food grains and small pebbles.

 

The herbicide hits chick embryos
hardest when their nerve cells are
beginning to link up

However, the exerted these
terat?ogenic e??ects on behaviour only
when the chick embryos received the dose
on the 15th day of incuhan'on?giving
on the 8th day of incubation had
no statistically signi?cant eflect. On the
8th day of incubation, the chick embryo
is rapidly forming its nerve cells. By
the 15th day, this process is nearly com-
pleted and the nerve cells are making
connections?synapses?with other nerve
cells. Sanderson and Rogers say ?the
brain therefore seems to be most sensi-
tive to at a time of maximum
synaptic formation". 

The scientists admit that they do not

know whether the il?a or it;
dioxin contaminant causes the amage_
However, the they administened
contained less dioxin than commercial
Australian (But not British 
which cannot. by law, contain more than
0-01 of dioxin.) Spraying Australian
on blackberries at the recom
mended rates of application, the re
searchers say, could expose the eggs 0:
wild birds to 22 mg/kg?higher than tbi
smallest close they found to damage the
unborn chick. As the chemical passe:
through the shells of eggs, Sanderson
and Rogers conclude that ?it may there
fore present a real risk to bird species";

Because the formulation of 2,4,5-1
sprays in Bn'tain is different, doses to the
eggs of British wild birds from 
would be smaller?about 2 mg/kg. But
the Australian researchers point out that
Some individual chicks were eSpeciall)
susceptible to the herbicide.

They believe that humans may be at
risk too. They cite previous, inconclusive.
research by Barbara Field and Charles
Kerr of Sydney University which showed
that the yearly birth rate of babies with
defem in their neural tubes in New
South Wales correlated well with how?
much farmers used in the previ-
ous year. What is more, the highest rates
of these abnormalities occurred nine
months after the peak spraying periods
in the antipodean summer. This evidence
?together with the rough estimate that
humans are three. time: more sensitive
to than chicks and rats?leads
Sanderson and Rogers to warn that ?our
?ndings therefore imply a risk to th
human species". 

 

A plant cell that can store electricity

Biochemists at Ohio State University
have combined "living" cell components
with biochemicals to make what
is e?ectively a giant specialised leaf cell
that makes and stores electricity with
an ef?ciency approaching what would be
practically useful. Chemical and biologi-
cal systems cooperate to make the new
cell work this way.

Elizabeth Gross and her colleagues at
Ohio ?rst made successful plant-based
photocells from particles carefully isola-
ted from chloroplasts?the organelles re-
sponsible for inside liv-
ing cells. New Cross, Rong Pan, and
Ravindra Bhardwaj have used the same
chemical support system to drive cells
based on whole chloroplasts, whichare
easier to separate from the rest of the
plant tissue (Nature, vol 289, 396).

The researchers deposited the chloro-

plasts onto a micro?lter membrane sep- -

arating two solutions. One solution con~
tained an electron-donating chemical,
and the other contained an electron ac.
ceptor. When light is shone through the
acceptor solution onto the chloroplasts,
both become excited, and electrons flow



2 Cell

plasts into the acceptor solution. Because
the acceptor molecule that they used is
sensitive to light, the scientists found
that their cell produced a voltage even
when they used dead chloroplasts. Taking
into account the proportion of the total
membrane area covered by chloroplasts,
the researchers calculate that a maxi-
mum of about 3 per cent of the total
light power is converted instantaneously
into electric power; the chloroplasts

supply about two-thirds.

There are two important factors which
make these ?gures a rather poor measure
of the new cell's performance. First, it
is dif?cult to measure exactly how much

 

the chloroplasts contribute to the cell
ef?ciency, because the biological and
chemical systems in the cell probably
act synergisticalIy?that is, the whole is
greater than the simple sum of the in-
dividual contributions. Second and
much more important the cell also
stores large amounts of the light energy,
so that instantaneous power-conversion
ef?ciency measurements are misleading.

Gross and her colleagues found that
after about half an hour's illumination,
the cell went on producing much the
same voltage in the dark for another
hour. Cells that can store solar energy in
this way are obviously much more usaful
than conventional photocells which only
produce electricity when illuminated. 

Cells based on
whole chloroplasts
have been used to
'convert the Sun's
enefgy into
electricity. Port of
a chloroplast,
72 000, is shown.
The plant?s use of
light energy begms
in the thin

1.33m v.
l2-March 1982 

Volume 215, No. 4538 i-

. - I
. . -.-. . u: 

 

 

 

LETTERS Nuclear Weapons Materials: T. B. Cochran. M. M. Hoenig, W. M. Arkin:
Assessmg Agncultural Research: W. W. Kan/tie: Transamazon
Highway: Impact on Indians: D. C. P. Grossman; X-ray Holography:
D. Sayre; The Cray-l: W. R. Smith; Vaccines: 
H. Langbehetm; Damaging Stereotype: B. Hansen . . . . . . . . . . . . . . . . . . . . . 1344 
0
U1
EDITORIAL Energy and Chemicals from Trees . . . . . . . . . . . . .. 
. . . . . . . . . . . . . . . . . . . . . ?w 1349 I
?13?
- ?we  -- 'r 
AMERICAN ASSOCIATION FOR THE ADVANCEMENT  OF. SCIENCE 
- -- - - - n-?-I-hle . .-. . - am?.

(A)
REPORTS Recent Volcanic Activity at Glacier Peak: J. E. Beget . . . . . . . . . . . . . . . . . . . . . . . . . . 1389 
Kinetics of Deligni?cation: A Molecular Approach1390
Dating of a Fault by Electron Spin Resonance on lntrafault Materials:
M. Ikeya. T. Miki, K. Tanaka . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 1392
ag-Adrenoceptor-Mediated Hyperpolarization of Locus Coeruleus Neurons:
Intracellular Studies in vivo: G. K. Agitajanian and C. P. VanderMaelen .
Oxytocin Receptors and Human Parturition: A Dual Role for 
Initiation of Labor: A.-R. Fuchs Rapid Electronic Auto?uorography ofI
Dimensional Gels: J. 3. Dr" 
Human Liver 
with . . . . . . . . . . . . .. 1413
-uect on Life-Span
and R. L. Walford . . . . . .. 1415
-cu Retinotectal Projection During Chick
_.L00n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 1418
steroid Regulation of Muscle Degeneration in an Insect:
M. Schwart: and J. W. Truman . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 1420
Technical Comments: Jumping Chickens: Relevance to Hazard in Humans:
M. L. Leng: L. J. Rogers and C. A. Sanderson . . . . . . . . . . . . . . . . . . . . . . . . . .. 1421
PRODUCTS AND Personal Computer: Cell Culture Chamber: Refrigerators: Sample Preparation: 
MATERIALS Human Cell Growth Factor: Infrared Spectrometer; Literature . . . . . . . . . .. 1424

Jumping Chickens: Relevance to Hazard in Humans

SCIENCE. VOL. 215. 12 MARCH l982

Sanderson and Rogers have reported
the results of a study on the behavioral
effects in chickens given "environmen-
tally relevant doses" of 
phenoxyacetic acid and they
postulated that humans would be three
times more sensitth to this herbicide
than chicks (I). The conclusions drawn
from this study do not re?ect a va.id

Copyn'ght 0 1982 -

evaluation of the data because the
doses were not environmentally relevant
for bird eggs. much less for humans. and
chick embryo data are not a suitable
model for safety assessment in humans
and are not used by any government
agencies for this purpose.

Recent studies by Lavy et ul. (2) have
shown that the maximum exposure to

18533

2.4.5-T likely to be encountered in hu-
mans, is {About mg per kilogramor
body weight. which is orders of magni-
tudes less than in the chicken Studies.
Furthermore. such exposure would be
likely to occur only in applicators who
are using knapsack sprayers and wearing
contaminated clothing or in mixers or
loaders who are not careful to avoid
contact with the concentrate and spray
solutions. Measurable exposure to 2,4,5-
is extremely unlikely in bystanders or
the general population (3).

1n the study of Sanderson and Rogers,
the doses ranged from 7 to 53 mg of
2.4.5-T per kilogram of body weight in-
jected into the yolk sac offenile chicken
eggs incubation.
and from 75 to 225 mg/kg injected intra-
peritoneally into chicks on day 2 after
hatching. The 2.4.5-T contained 0.03
part per million of the highly toxic con-
taminant 2.3.7.3-tetrachlorodibenzo-p-
dioxin (TCDD). which is representative
of the level in 2.4.5-T produced world-
wide today.

"Environmental relevance" was
based on an estimated dose of 22 mg/kg
for an egg contaminated with 1.1 mg of
2.4.5-T over the entire shell surface
(area, 50 cmz). presumed to be equiva-
lent to Spraying an 80 percent 2.4.5-T
product (derivative n0t speci?ed) at a
rate of 2.8 liter/ha. The latter is equiva-
'ent to 224 mg of (or derivative)
per square meter of horizontal surface
area. not the entire egg surface. and
2.4.5-T is generally applied as a 2 to 5
percent solution in oil, oil-water. or wa-
ter emulsion. Injection of2.4.5-T into the
egg was also assumed to be equivalent to
contamination of the shell surface. How-
ever. only a small fraction of the herbi-
cide applied to the surface of an egg
penetrates the shell and reaches the
chick embryo (4).

Sanderson and Rogers concluded that
humans would be at least three times as
sensitive to 2.4.5-T exposure as rats or
chickens. This conclusion was based on
Studies by Piper er al. (5) and Erne (6),
although neither study implied such a
relationship. What happens as a result of
the injection of a large dose of a chemical
into an isolated egg is not representative
of what might happen in a human fetus
connected to its mother?s ef?cient detox-

i?cation system. particularly for a chem- 

ical such as which is rapidly
excreted in the urine of humans (7).
Even more questionable is their extrapo-
?ion of effects from the injection of a

I422

nearly lethal dose of 2.4.5-T into newly
hatched chicks to the potential effects
from low-level exposure in adult hu-
mans. Unfortunately. reports of such
studies done under highly exaggerated
conditions continue to add to the confu-
sion surrounding the use of bene?cial
agricultural chemicals such as 
MARGUERITE L. LENG
Health and Environmental Sciences.
Dow Chemical ontpany.
Midland. Michigan 48640

References and Notes

I. C. A. Sanderscm and L. J. Rogers. Science 21!.
593 (I981).

. T. L. Lavy. J. S. Shepard. J. D. Maltice. J.
Agric. Food Chem. 28. 626 (I980): T. L. Lavy.
S. Shepard. D. C. Bouchard. Bull. Environ.
Contam. Toxicol. 24. 90 ?980): T. L. Lavy.
paper presented at the meeting of the Weed
Science Society of America. Toronto. Canada.
February I980.

3. M. L. Leng. T. Lavy. J. C. Ramsey. W. H.
Braun. in Pesticide Residues and Erposure
(Symposium Series l82. American Chemical So-
ciety. Washington. D.C.. 1982). pp. 133-l56.

4. J. D. Somers. E. T. Moran. Jr.. B. S. Reinhan.
Bull. Environ. Contam. Toxicol. ll. 5? H974).

5. W. N. Piper. J. 0. Rose. M. L. Leng. P. J.
Gagging. Toxicol. Appl. Pharmacol. 26. 339
(I 

. K. Erne. Am: Vet. Scand. 7. 240(1966).

. P. I. Gehring. C. G. Kramer. B. A. Schwetz. J.
O. Rose. V. K. Rowe. Toxicol. Appl. Pharma-
col. 26. 352 (1973).

30 March l98l: revised June I981

IJ



Several aspects of Leng?s technical
comment require correction. We found
that. for chicken eggs. the sensitive
period to 
acid was day IS of incubation.
when the injection of as little as 7 mg per
kilogram of body weight caused subse-
quent behavioral abnormalities (1). This
amount does not cause marked morpho-
logical abnormalities and is well below
the LDSO of 53 (the dose lethal to
50 percent of the animals teStedi [see
(I ll.

The question of the amount ol? 
absorbed through the eggshell is. of
course. controversial. If we accept that
only 30 to 50 percent of an egg may be
covered by the spray and if we use the
concentration recommended by the gov-
ernment of Victoria. Australia. for
spraying blackberries, an egg would re-
ceive 7 to II mg/kg. Even ifonly some of
this is absorbed. it is still uncomfortably
close to the amounts that caused behav-
ioral abnormalities in our sample. Leng
does not allow for a safety factor. At-
tempts to extrapolate from these results
to the calculation of the ?no-effect" dose
in humans are complicated by individual
variations in sensitivity to 2.4.5-T [see
ourjumping data in 

We used the acid form of 2.4.5-T,

which is less toxic than its esters. usually
used for Spraying. Moreover. we gave
only one dose to each egg. Repeated
dosing may well be toxic at lower daily
rates of administration. Leng quotes a
maximum exposure to 2.4.5-T of mg}
kg for knapsack Sprayers but fails to say
that this value was calculated for one
Operation lasting 180 minutes. Most
Sprayers are repeatedly exposed and for
longer periods. The maximum exposure
measured by Lavy et al. (2) was actually
L85 mg/kg per Spraying operation. and
this did not include spray inhalation.
which is probably a more important
route of exposure. particularly in the
vicinity of aerial Spraying.

We agree that other species must be
tested with 2.4.5-T. We have in fact
shown that the orai dosing of pregnant
rats with as little as 3 or 6 mg/kg on day
8 of pregnancy causes behavioral ab-
normalities in the pups tested several
months later (3). This dose is one-hun-
dredth of the L059 in this species and
one-tenth of the lowest dose reported to
cause physical deformities (4). Phenoxy-
acetic acids themselves appear to be able
to cause these eti'ects; we have found
similar e?'ects with 2.4?dichlorophenoxy-
acetic acid which does not con-
tain 
(TCDD) (3).

Our statement that humans are likely
to be more Sensitive than rats and chicks
is based on deductions from the plasma
which tor an oral dose oil-1.5-
of5 tag/kg is 23.] hours in humans and
4.7 hours in rats (.5). The results for
chicks appear to be similar to those for
rats (5). We therefore maintain that
2.4.5-T (and 2.4-0) may present risks to
brain development and function in hu-
mans and other species.

L. J. ROGERS
A. SANDERSON
Pharmacology Department.
Monash University.
lay-ton. Victoria 3168. Australia

References

l. C. A. Sanderson and L. J. Rogers. Science 2l 1.
593 (I981).

2. T. L. Lavy. S. Shepard. D. C. Bouchard.
Bull. Environ. Contam. Toxicol. 24. 90 I930).

3. At. A. Crampton and L. J. Rogers. in prepara-
tion: P. J. Booth and L. J. Rogers. in prepara-
tton.

4. K. S. Khera and W. P. McKinley. Toxicol.
Appl. Pharmut'ol. 22. I4 (I972). 

5. P, J. Gehring. C. G. Kramer. B. A. Schwetz. I.
Q. Rose. V. K. Rawe. ibid. 26. 352 (1973): W.
N. Piper. J. 0. Rose. M. L. Lang. P. l. 
ibid.. p. 339'. M. W. Sauero?'. W. Bruun. G.
E. Blau. l. E. LeBeau. ibid. 37. Iin (1976); K.
Erne. Aria Vet. Scand. 7. 240(1966).

23 Noyembet 

16534

SCIENCE. VOL. 2l5

sou

 

2898' lZ

 




EPIDEMIOLOGY OF SOFT-TISSUE SARCOMA
AND RELATED HUMAN RESEARCH

(as related to herbicide exposure)

LAWRENCE B. HOBSON, M.D., 

Deputy Director
Agent Orange Projects Office
Veterans Administration
March 1983

?mmeu



?org
?Aerknude
SOHI h?si

S?mrcam

999azo?n

BVQUZIZ ?00?

16535 

EPIDEMIOLOGY OF SOFT-TISSUE SARCOMA
AND RELATED HUMAN RESEARCH

Initiation of Swedish Studies of Herbicides and Cancer

In 1972 Swedish newspapers published rumors that rail- -
road workers were dying from lung cancer as a result of
exposure to herbicides used in their work. The National
Board of Occupational Safety, as a result, requested
Professor Olav Axelson, a specialist in occupational medi-
cine, to undertake an epidemiological investigation of the
matter.

The results of this investigation have been reported in
a series of four papers, 174. Another series, 5-7, was
prompted by criticism of the epidemiological and statistical
methods employed in this and related studies. Attention in
the United States has focussed on the pre-publication manu-
script of the 1980 paper by Axelson, Sundell, Andersson,
.Edling, Hogstedt, and Kling, i.

This paper dealt with two aspects of the study of rail-
road workers. The initial phase was a cohort study of 348
men who had been exposed, individually rather than as a
group, to herbicides for more than 45 days during 1957 to
1972 and who were followed through October 1978. Exposure
information was incomplete but the workers were divided into
subcohorts with exposure to phenoxy acids (which include the
ingredients of Agent Orange), amitrol, or to both herbicides.
The mortality rates for these exposed subcohorts were com-
pared to the age-specific national death rates for Swedish
men, the latter serving as the control cohort.

Overall 49 deaths were expected in the exposed cohort;
45 occurred, a result attributed to the "healthy worker
effect." There were, however, 17 tumors found where 11.85
were expected. Among the deaths occurring at least ten years
after the first exposure, 6 cancers were found although only
1.78 were expected. Dr. Axelson, 8, later increased this to
7 tumor cases. Each subcohort had an excessive number of
tumor deaths, the greatest being in the group exposed to both
phenoxy herbicides and amitrol.

Although initially, 2, amitrol was associated with an
increased tumor mortality: somewhat different results were
found in a second phase of the examination, described as a
case-referent study (identical to a case-control study).
data indicated a "statistical association" of phenoxy
herbicides and excess tumor mortality, 3. Suspicion was
increased by finding that workers exposEd to phenoxy acids

The

0990212 H00

16536'

alone had a "statistically significant excess of stomach
cancer", specifically 2 cases compared to 0.33 expected when
this type of herbicide was used alone and 3 cases as compared
to 5.1 expected Jincreased from 4.1, 8) for all workers
exposed to phenoxy herbicides, alone 6: with amitrol.

The series of papers on railroad workers has been criti-
cized on several methodological grounds, 9, and Axelson has
replied to these criticisms, 6. Richard 5} Remington, Dean
of the School of Public Health, University of Michigan, re-
viewed this and other Swedish studies for the Office of Tech-
nology Assessment. His evaluation, 10, was that the Axelson
investigations had been "carefully and "well re-
ported." He pointed out the limitations of the statistical
methods used and found that "the numbers available are
inadequate to permit definite conclusions" although "the
results are suggestive."

Of interest in connection with the question of soft-
tissue sarcomas and phenoxy herbicides is the type of tumors
found by Axelson's group, 1, One case of Hodgkin's 
occurred among the eight tumors in men exposed to phenoxy
herbicides alone and no soft-tissue sarcomas or non-Hodgkin's
was diagnosed among the eight tumors appearing in
workers exposed to both amitrol and phenoxy herbicides. In
other words, no sarcomas were reported for the total of 207
men exposed to phenoxy compounds, 2. A reticulum cell sar-
coma and a Hodgkin's were?found among the 7 tumors
of workers exposed to amitrol alone. Thus, there was one
soft?tissue sarcoma reported for 152 men exposed to amitrol,
3. Another 28 persons described as exposed to "other herbi-
cides and combinations" cannot be idenzified as to exposure
to specific herbicides but apparently none developed a tumor.

1990212 H00

Axelson's work is directly related to the later work on
soft-tissue sarcomas and by Lennart Hardell. In-
deed, Axelson suggested to Hardell in 1976, that he conduct a
case-control study of soft-tissue sarcomas and has actively
assisted in Hardell?s work since then.

Swedish Investigation of Soft-Tissue Sarcoma

That work began when Hardell admizted for treatment 3
patients in the autumn of 1976, each with a soft-tissue sar-
coma, and a history of exposure to phenoxy herbicides. He
then found a total of 7 patients with 'malignant 
tumors" (soft-tissue sarcomas) who gave a history of having
worked with phenoxy herbicides 10 to 23 years earlier. The
cases were among 87 patients with soft-tissue sarcomas, 55 of

2 - 16537

whom were men. Of these men, 9 were forestry workers, 6
worked in forestry and on farms, and 6 were employed in saw
mills or pulp plants. Another two tumors appeared in men
whose connection with forestry was less direct. The malig-
nancies found were two leiomyosarcomas, two rhabdomyosar-
comas, two neurofibrosarcomas, with one each of fibroid 
liposarcoma, myxofibrosarcoma, and polymorphocellular


Following Axelson's advice, Hardell began a case?control
study that was published in two journals, 13, 14, a common
practice of reporting in Swedish with an almost?identical
paper in English. The second paper in English aroused much
interest in the United States.

Hardell and found 21 living and 31 dead men
who were diagnosed as having soft-tissue sarcomas in
Hardell's oncology department in northern Sweden. They were
matched for age and place of residence, as well as date of
death for the deceased, with other men selected from the
Swedish National Population Registry or from the National
Registry for Causes of Death. Each living patient had 4
living controls; each dead man had 4 deceased controls. Ex-
posure information was sought by the use of a mailed ques-
tionnaire that has never been published. It contained 130
questions, including 16 about the use of organic solvents, 4
about plastics, 3 about glues, 4 about drugs, "several" about
smoking habits and an unstated number about exposure to phe?
noxy herbicides and chlorophenol as used in the lumber and
paper mills. This questionnaire was mailed to the patient or
to his next of kin if he were dead, 12:

When the answers to the questionnaire were less than
clear, a supplementary interview was obtained, usually by
telephone, with the interviewer unaware of the health status
of the person in question. Employers, neighbors, and others
were consulted "if necessary to verify and monitor the accu-
racy of the exposure information," 

2990le H00

Using the criteria for exposure established for the
study, 36.5% of the 52 patients and 9.2% of the 208 controls
had been exposed to phenoxy herbicides and/or chlorophenols.
The "relative risk" of developing soft?tissue sarcomas was
calculated as 5.7, i.e. men exposed to the chemicals had
almost six times as great a change of developing a sarcoma as
did those who were not exposed. The relative risk was 5.3
for the 46 men exposed to phenoxy herbicides alone, and 6.6
for the 40 men exposed only to chlorophenols. It was thought
that confounding factors had an insignificant effect.

16538.

The authors concluded that "the investigation showed an
increased risk for soft-tissue sarcomas? but ?a Specific
evaluation of the effect of separate chemical substances was

not possible," 13:

The study's methods have been criticized and doubts have
been expressed about the 100% response rate to the question-
naire approach, 9. (Actually, 2 of 208 controls did not
answer, 15:) The statistical approach was described as
misrepresented and a major criticism was leveled
because of the possibility of "selective recall," the greater
tendency for an ill person to remember a supposed "cause" for
the illness than a well person would have to remember the
same "causal" event.

The criticisms evoked several replies. Axelson defended
the case?control design, the objectivity of obtaining expo-
sure data retrospectively, and the statistical techniques, 5.
He concluded that the use of interviews for determining ex-_
posure is justified, 1, and defended in principle the treat-
ment of confounding factors, 16. Hardell recalculated the
1979 results and his subsequent papers to substantiate his
earlier findings and performed a separate investigation in
support of his confidence that "no substantial observational
bias could exist in the studies," lg,

Remington, 123 expressed the view that "the findings of
this particular investigation are suggestive'I and that "a
relative risk of 5.3 for exposure to phenoxyacetic acids must
be taken seriously." HOwever, "case-control studies are
uniquely susceptible to hidden sources of bias" even when the
investigators are "unusually careful" as they are in this
"excellent investigation."

EQQOZIZ MUG

Hardell's group also undertook a second case-control
study of identical design in southern Sweden which is more
devoted to agriculture than to forestry, 11! lg. In this
investigation each of 72 living and 38 dead patients was
matched with two controls. Among the 110 cases, 22.7%
reported exposure to phenoxy herbicides or chlorophenols and,
among the 219 controls, 5.9% were so exposed. -This gave a
relative risk of 5.1 with matching and 4.7 when the matching
was dissolved, i.e. when sorting by age was ignored during
statistical calculations. The relative risk from exposure to
phenoxy herbicides was calculated to be 6.8, and that from
chlorophenols to be 3.3. Exposure to more than a dozen other
noxious materials, e.g. asbestos, smoking, DDT, and lindane,
were considered as possible confounding factors although none
was found to be clearly associated with an increased risk by

itself.

18539

The reports list the diagnoses of all 110 cases of soft~
tissue sarcoma as: leiomyosarcoma, 33; malignant fibrous
histiocytoma, 19; liposarcoma, neurogenic sarcoma, 11;
angiosarcoma, 9; myxofibrosarcoma, 7; fibrosarcoma, 5; derma-
tofibrosarcoma, 3; atypical fibroxanthoma, synovial sarcoma,
sarcoma NOS, 2 each; Ewings's sarcoma (extraskeletal) and
rhabdomyosarcoma, 1 each. No statement is made as to which
of these tumors was found in the 25 cases with identified
exposures and no histological diagnoses are reported for the
northern Swedish series, 11, L1.

The authors of the southern Swedish study conclude that
"exposure to phenoxy acids and chlorophenols might constitute
a risk factor in the development of soft tissue sarcomas,"
lg. The investigation has been the subject of the same
criticisms and refutations as the earlier study.

Remington concludes that "the results are consistent
with the hypothesis that phenoxy acid exposure increases the
risk of tumors of this type" but adds that "case?control
methodology is intrinsically susceptible to subtle and un?
measurable biases."

Swedish Investigation of 

 

In May, 1978, Hardell was prompted to a new study by a
patient with a malignant histiocytic and a history
of "massive exposure to phenoxyacetic acids." All men admit-
ted to the oncology department with this type of tumor during
the first nine months of 1978 were asked about their occupa-
tion and possible chemical exposure. Of 17 patients, 14 re?
ported an occupation consistent with exposure and 11 of them
had had contact with phenoxy herbicides or chlorophenols ten
or more years earlier, 12:

11990212140?

These observations led to a case?control study, the
report of which in 1981, 31, differs considerably from that
in 1980, 20. The earlier report was commented upon in manu-
script de?'by various experts but.the later version will be
used here.-

The investigation, in collaboration with Axelson, 
included both Hodgkin's disease and non-Hodgkin 
The 169 cases consisted of 60 Hodgkin's disease patients
predominance, 20; nodular sclerosis, 3: mixed
cellularity, 27; depletion, 10), 105 men with non?
Hodgkin's (follicular center cell (FCC) type, 53;
non?FCC type, 52), and 4 individuals with unclassifiable
'Each case had two matched controls, 338 in all.
Of the cases, 62 had died as had 124 of the controls. 

16540~

Questionnaires and interviews were used to determine

I exposure to phenoxy herbicides, chlorophenols, organic
solvents, or medicines and to characterize jobs, hobbies, and
smoking as they were determined in the soft-tissue sarcoma'
investigations,_gg. All cases and controls were from
northern Sweden.

Cases in which exposure was reported to chlorophenol, or
to "mutagenic" solvents (benzene, trichloroethylene, perchlo-
roethylene and styrene) were divided into high-grade and low~
grade exposure groups. Continuous exposure for a week or
less or repeated exposures totaling less than a month were
considered low?grade. Analyses also divided cases into two
groups depending on whether 5 years had elapsed as a latency
period between the first exposure to the chemical and the
tumor diagnosis.

Of the cases, 36.1% had been exposed to phenoxy herbi-
cides or chlorophenols: 9.6% of the controls had been so
exposed. The relative risk for these exposures was 6.0 with
matching and 5.3 without it. Phenoxy herbicides gave a rela-
tive risk of 4.8 although it was greater if exposure was for
90 days or more. Chlorophenols gave relative risks of 8.4
for high?grade exposure, 2.9 for low?grade. High- and low-
grade exposure to organic solvents gave relative risks of 2.8
and 1.2 respectively. On the other hand the few cases with
both phenoxy herbicide and high?grade organic solvent expo-
sure was calculated to have a relative risk of 11.2 and some
other combinations also gave large relative risks. The
length of the latency period, however, seemed to have no

effect.

The authors conclude that "this investigation suggests
that exposure to organic solvents, chlorophenols, and/or
phenoxy acids constitutes a risk factor for malignant lym?
phoma," 21. Dr. Remington commented that "a substantial and
statistically significant relative risk is found for this
group of tumors. And again, pnenoxy acid exposure is
specifically incriminated." He continues, however, that the
limitations of case-control methods have to be considered as

well.

9990212 mm

Swedish Investigation of Carcinoma of the Colon

Hardell undertook to answer doubts that his question-
naire and interview methods allowed observational bias in
assessing exposure by conducting a case-control study of
"colon cancer." The condition is not suspected of having any
association with phenoxy herbicides or chlorophenols. In
consequence, if the previously used exposure determination

6 16541

resulted in a relative risk of 1.0 or near it, there had been

I no observational bias in the questionnaire?interview
procedure used in the earlier studies of soft-tissue sarcomas

and 

Of the 157 men with colon cancer all but 3 answered the
questionnaire. The controls consisted of the control groups
from the soft-tissue sarcoma study (206 men) and the malig-
nant study (335 men). In all, 41% of the cases and
45% of the controls were dead. Of the cases and controls,
11.0 and 10.4% reSpectively had been exposed to phenoxy
herbicides or chlorOphenols. For phenoxy herbicides, the
relative risk was calculated to be 1.3 and for chlorOphenol
it was 1.8. Neither was significantly above 1.0. The con-
clusion was that "the previously reported associations be-
tween exposure to phenoxy acids or chlorophenols and soft-
tissue sarcoma and malignant cannot to any essential
degree be explained by observational bias," 

Later Criticism of Swedish Studies

There remain, however, doubts about the practical
significance of the Swedish epidemiological studies stem-
ming from several of their characteristics. The main
criticism is the reliance on recall of the men or their
relatives, employers and associates for undramatic events
years earlier as well as the possibility of unconscious
bias on the part of the interviewer, the "observational
bias" discussed above. Coggan and Acheson point out that
the positive association between exposure to phenoxy
herbicides and the development of several or many soft~
tissue sarcomas, Hodgkin's disease and non-Hodgkin's
may indicate "a serious undetected bias" even
though the explanation has been offered that all these
tumors are embryologically related, 32, These authors
conclude that "it is as yet impossible to estimate with
any precision the risk of soft-tissue sarcoma due to
phenoxy.herbicides" but add that "there'is suggestive
evidence of a biological association between phenoxy
herbicides (or their contaminants) and soft?tissue sar-
comas." They feel that there is weaker evidence for an
association between herbicides and 

9590212 M00

Hardell and Axelson reject the idea of observational
bias, citing the colon cancer study as evidence, 31.
They also defend the aggregation of tumors because of the
"so?called addition theorum for chi-square and Poisson 
distributions" as well as the embryological relationship -.

of the neoplastic tissues.

16542

American Support for Swedish Conclusions

Support for the connection between soft-tissue sarcomas
and exposure to phenoxy compounds has been reported in
several papers from outside Sweden. The data most often
cited as favoring the relationship are derived from observa- -
tions in the American chemical industry.* The first was a
note by Honchar and Halperin in which they pointed out that
of 105 deaths in four exposed industrial "cohorts" 3 
were due to soft-tissue sarcoma, whereas only 0.07% of deaths
among adult American men are so caused. The three cases were
malignant fibrous histiocytoma, fibrosarcoma, and liposar-
coma. The authors felt that these ?suggest a common pat~
tern," 33, Cook added a fourth case, another malignant
fibrous histiocytoma and noted that all four were smokers and
two had 

Moses and Selikoff reported a fifth case, a non-smoker,
with neurogenic sarcoma (malignant schwannoma). They give
the total annual incidence of soft-tissue sarcomas as 4500
(less than 1% of newly diagnosed cancers) in the U.S. and
quote 4.9% of soft-tissue sarcomas as malignant schwannoma,
26.

Johnson and his co~workers briefly described a young
man who died of fibrosarcomatous mesothelioma some four
years after first being exposed to phenol. His father
had a liposarcoma after "prolonged exposure" in a plant
manufacturing chlorinated phenols among other chemicals,
27.

Hardell and Ericksson accepted the two additional cases
to total 7 deaths from soft-tissue sarcoma among 105 deaths
among American industrial workers, the expected number being
0.07%. This would "fit in with" the Swedish investigations,
they believe, 3g.

ZQQOZIZ mm

To date no critical review has been made of the cases
and the industrial population in which they were detected.
The reports have been brief "Letters to the Editor" and each
discusses one to three cases. The total of 105 deaths used
as the number of dead workers has not been kept current as
new soft-tissue sarcoma cases were added and the total number

 

*Data given by Honchar and Halperin, Cook, Moses and
Selikoff, and Johnson et al pertain to workers at Monsanto
Company and Dow Chmical Company. For details -of studies
of these workers see 24a and 25a.

18543

of exposed workers has not been given. No use has been made
of controls, even in the form of a retrospective cohort
comparison.

A case report without statistical data briefly described
three soft?tissue sarcomas among Vietnam veterans who re?
ported exposure to phenoxy herbicides in that country. One
man had an inflammatory histiocytoma, another suffered from a
fibrosarcoma, and the third had a leiomyosarcoma,_gg.

European Suoport for Swedish Conclusions -

Barthel determined the frequency of malignant neoplasms
among 1791 pesticide sprayers and agricultural technicians in
East Germany during 1976 to 1979. He states the retrospec-
tive cohort study used police as controls but gives no data
for them. After eliminating "on statistical grounds" 133
cases who died before 1970, be compared the mortality rate
and cancer incidence with corresponding figures from the
death statistics and the cancer registry of the Health and
Social Welfare. The "case" group had multiple exposures ove:
the years to fungicides, insecticides, and herbicides in-
cluding phenocyacetic acids. Among 169 malignant neoplasms
in 1658 exposed men were 1 3 plasmacytomas, 1
described as a malignancy of tissue, and 1 of soft-
tissue, not otherwise characterized. Bronchogenic carcinoma
was the most common malignancy with 59 cases, double the
expected occurrence, although the cases had smoking habits
like those of the general population, 30. A brief report
describes a case of non-Hodgkin?s and a second of
malignant among 158 workers with pentachlorophenol.
This type of neoplasm would have an expected occurance of

0.28, E.

Studies Not Supporting Swedish Conclusions

In contrast to the reports of an association between
phenoxy herbicides or related compounds and soft?tissue sar-
comas and malignant some investigators have found
no association. Some of these investigated a possible rela-
tion, others were "follow?up" studies of industrial workers
in whom no sarcomas or were found.

Dr. Riikimaki and his collaborators have completed nine
years of mortality study following 1,926 persons who worked
with phenoxy herbicides in Finland during the 1955-1971
period. All had at least
of the men totalled eight
mortality rates among the
national death rates. As

weeks or more as of 1971. The
workers were compared with the
of 1980, there had been 82 deaths

two weeks of exposure and a quarter-

85902121400

167544 .

of exposed men as compared to 91 expected and, of these, 17
were cancer deaths with 18.4 expected. There were no cases
.of soft-tissue sarcoma nor of although 0.1 and 0.8
would have been expected. The authors believe that "the
investigation cannot be regarded as a conclusive negative
study" but point out that the "results do not confirm the 
association between mixed herbicide exposures? and cancer
risk, 23:

Hogstedt and Westerlund compared the mortality rate of
Swedish supervisors and workers in forestry. The supervisors
were fewer in number (142) than the workers (244) but the
former were judged to have been more heavily exposed. The
relative risk of death was about the expected but, after a
10-year latent period, the relative risk for cancer was about
4 for the supervisors and only about 0.4 for the workers.

The fatal tumors were of various types but there was no soft-
tissue sarcoma or -

Two case-control studies in New Zealand have been initi-
ated by Smith et al to examine the association suggested by
Swedish studies of phenoxy herbicides with soft-tissue sar-
comas and malignant In the first investigation,
102 cases of soft-tissue sarcoma have been identified in men
from the New Zealand Cancer Registry between 1976 anmd 1980.
An equal number of matched controls with other forms of
cancer were selected for comparison. The sarcomas are fibro-
sarcomas, 25; liposarcomas, 20: rhabdomyosarcomas, 9; leio?
myosarcomas, 7; malignant histiocytomas, 6; other types, 22;
and unspecified, 13. The preliminary report compares cases
and controls as to the occupation shown on the Registry en-
rollment.: There was no significant difference betwen the
groups as to the number of men working in agriculture, fores-
try, and fishing, the occupations with the greatest likeli-
hood of exposure to phenoxy herbicides and chlorOphenols.

The only occupations associated with soft-tissue sarcomas
exclusively are blacksmiths,_machine tool operators, electri-
cal fitters, and electrical workers. The investigators are
now obtaining work histories for cases and controls by tele-
phone interviews and warn that later results may change their
conclusions. The data at present "do not give evidence for a
relationship (of soft?tissue sarcoma) with occupational expo-
sure to phenoxy herbicides and chlorophenols" but "should not
be taken as substantive evidence against the hypothesis", 33,

A second report by Smith et al includes the results of
the telephone interviews regarding 80 cases and 92 controls
already completed. Probable or definite exposure to phenoxy
herbicides for more than one day earlier than five years
before cancer registration was found in 17 cases and.13 con-

10

5590212 H00

16545

trols, giving an odds ratio of 1.6. This would be expected
I to increase when the exposure criteria were more stringent
but, when exposure was at least five days and more than ten
years before registration, there were 13 cases and 12 -
controls included reducing odds ratio of 1.3. Neither ratio
is statistically significant and there have been no
soft?tissue sarcomas reported among the most highly exposed
group of 2000 aerial and ground sprayers. The results, the
authors believe, "do not generally support the hypothesis
that exposure to phenoxy acid herbicides cause softntissue
sarcoma," 22.

A brief initial report by Edling and Granstam compared
the causes of death for 375 Swedish forestry workers, aged 25
to 69 years, who died during 1968 to 1977, with the mortality
figures from the Swedish national statistics. There were 75
deaths from all malignant tumors, as compared to 86 expected.
Renal tumors killed 8 with 3.84 expected and "tumors of lym-
phatic and hematopoetic systems" were responsible for 14
deaths with 7.5 expected. No deaths were attributed to soft-
tissue 

In addition to these studies, several small industrial
groups have been followed well into the latent period for
solid tumors. None has been reported to include cases of
soft?tissue sarcoma or malignant Hay examined 41
of 79 workers who developed chloracne following accidental
exposure to trichlorophenol in 1968 at the Coalite Company in
Great Britain. Another 54 employees were possibly exposed.
None of the workers had significant changes ten years after
the accident and neither death from nor evidence of neoplasm
was found, 37. Jirasek's group has closely followed 55 men
who were intensely exposed during the manufacture of 2,4,5-
trichlorophenoxyacetate from 1965 to 1968 in Spolana, Czecho-
slovakia, and who developed evidence of acute intoxication.
Two workers died of bronchogenic carcinoma 5 to 5.5 years
after the first exposure. There was no other evidence of
malignant neoplasms during a ten?year 



In 1963 an explosion at Philips?Duphar, Amsterdam, ex-
posed 106 workers involved in manufacturing 2,4,5-tetrachlo-
rophenyoxy acewtate. Among the 93 workers followed to 1977,
only one death 14 months after the accident was due to cancer
and the pancreatic carcinoma involved was apparently symto-
matic before the explosion. No case of soft-tissue sarcoma or
malignant was reported, 22:

One study is often cited with the Swedish studies al-
though it did not deal with soft?tissue sarcomas and malig?
nant 40. A more recent review by Thiess et al
reports that elf?74 exposed persons are still being followed

116545

after 26 years. There have been 21 deaths, about equal to
the 18 to 20 deaths expected from major comparative popula-
tions and 18 and 19 deaths expected among matched unexposed
controls. Cancer was responsible for 7 deaths as compared.to
4.1 expected from the comparative populations and 5 in each
internal control group. Gastric carcinoma in 3 exposed
persons exceeds the expected 0.61 to 0.70 expected cases.
There were, however, no soft-tissue sarcomas or malignant
among these chemical workers at BASF, 41.

A number of other industrial exposures to phenoxy herbi?
cides, their precursors or contaminants were reported before
1973, 42. The pOpulations were small but generally heavily
exposedT Unfortunately it has not been possible to locate
late reports on the exposed populations although ten years or
more have elapsed since exposure.

The accident at the ICMESA factory in Seveso, Italy, in
July 1976 exposed many people to trichlorphenol; more than
5400 adults and children of,both sexes are known to have been
in contact with the chemicals for several days, 43. Although
only about six years have elapsed since the exposure, the
population has been under surveillance and the rate and
causes of death are being followed. To date no soft-tissue
sarcomas or malignant have been reported.

Another less systematic observation bears on the situa-
tion. The phenoxy herbicides have been used frequently and
extensively in agriculture and forestry in the United States
since the late 1940's. They were used on lawns in cities, as
well, for most of that period. If the relative risk of de-
veloping so distinctive a group of tumors as the soft-tissue
sarcomas and the malignant had increased by 5 of 6
fold over that before 1945 as the Swedish studies would pre-
dict, it almost certainly would have been evident to clini?
cians and pathologists, especially in the rural areas, even
without systematic studies. No such increase was noted.

Critical Evaluations

 

The Swedish investigators have been cautious in inter-
preting their results. In his medical dissertation based on
his epidemiological studies, Hardell judges that the similar
results in the two case~control investigations ii,
16) "seem to increase the confidence that the observed asso?
Eqation of exposure to phenoxy acids and soft-tissue sarcoma
was not spurious" and did not believe that confounding fac-
tors "could acc0unt for the observed relation." In summary,
he concluded that "it is suggested that exposure to phenoxy

12

IQQUZIZ HUG

16547

acids should be looked upon as an occupational cancer
hazard,"lii.

Other reviewers have been more skeptical as to the sig-
nificance of the work. Remington's overall opinion was that
"in toto, the Swedish work is'credible if not fully conclu-
siVe. Certainly this work would seem to justify further
investigation," 15% 'Coggan and Acheson, after reviewing
other work as well as the Swedish studies, state that "on
the present evidence it seems possible that soft-tissue sar?
comas have arisen in association with exposure to phenoxy
herbicides" but continue that Fit is as yet impossible to
estimate with any precision the risk of soft-tissue sarcoma
due to phenoxy herbicides." They conclude that "there is
suggestive evidence of a biological association between
phenoxy herbicides (or their contaminants) and soft-tissue
sarcoma. The evidence relating these products to the
occurance of is weaker," 22. An unsigned editorial
in Lancet commenting on the Opinion? of Coggan and Acheson
seems to agree with their conclusions with regard to
soft-tissue 

 

Hardell and Axelson disagreed with both the Coggan
and Acheson's opinions and the Lance: editorial, 23.
They have been at some pains to counter charges of-"ob-
servational bias," lg! but have not convinced everyone
that faulty memories do not result in significant errors
in evaluating 

The causal connection between phenoxy herbicides and
soft?tissue sarcomas would be much more likely if there
were a unique preponderance of one type or even of a few
types in the exposed men. The Swedish reports never
compare the morphological types or location of the malig-
nant tumors in cases with those in controls, 52. Their
only justification for aggregating the types, and presum-
ably for omitting the data from their reports is "the
uncertainty of relations between the various histological
groups in terms of causal mechanisms" and "the so-called
addition theorem for chi-square and Poisson distribu?
tion," 23. The uncertainty of causal relations is
precisely the reason for reporting the groups and the
addition theorem cannot justify the aggregation of unlike

 

has been suggested
as the principal carcinogen in the phenoxy herbicide
and trichlorophenols but this has been disputed.
See 22, 23, 46. The controversy is not considered

in this discussion.

13

3990212 mm

1-6 5.138

entities unless significant common factors have been
demonstrated.

Scientific results are strengthened greatly when
independent investigators substantiate them. The Swedish
studies have been said to be independent and confirma- 
tory. The two soft?tissue sarcoma investigations do
support one another L12, 13, 14, 16) but they are the
work of the same group of_ihvestigators. The investiga-
tion of malignant histiocytic was also conducted
by the same group but was a case-control study of a
separate entity, Axelson's work on herbicide
exposure and cancer was not truly independent from
Hardell's efforts since Hardell has recognized his in-
debtedness to Axelson for his assistance in the first
case-control study, (13, ii). More important Axelson did
not associate phenoxy herbicides or chlorophenolic com-
pounds with soft-tissue sarcomas nor with malignant lym?
phomas among railroad workers, 1-3,

The reports of soft-tissue sarcomas among chlorphenol
workers in the United States (24-27) have been cited as
supporting Hardell's conclusioEEZ?2l, 28, 44, 46. The data
have been reported piecemeal without.a?Elearly?Ehumerated
total population from which they were drawn. The comparison-
was made to mortality data for the general population of the
appropriate age and sex. The type of soft-tissue sarcoma is
known for each case; among the 7 men were 2 malignant fibrous
histiocytomas, 2 liposarcomas, as well as one each of fibro-
sarcoma, malignant schwannoma, and fibrosarcomatous mesothe-
lioma. As before, the tumors are not of a uniform type.

8990212 H00

Coggan and Acheson comment that the Swedish studies and
the American reports taken separately do not "provide con-
vincing evidence that the incidence of soft?tissue sarcomas
is increased after exposure to phenoxy acids and chloro?
phenols, -- Considered together the whole becomes more
persuasive." They add that "it is surprising that the asso-
ciation should apply to tumors of such a variety of tissues,?I
22. The Lancet editorial finds only that "the number of
deaths due to soft-tissue sarcomas [in the American data] is
disturbing," ii:

In addition to the American experience, the British
(37), EurOpean (30, 32, 33, 36, 38-41) and New Zealand (25?
337 medical and scientific-writers'haye studied populations
fiye years or longer after exposure to phenoxy herbicides
and/or chlorophenols in a variety of situations, some intense 
and acute, others prolonged. Only one observer (39) reported
a case described as a soft-tissue malignant neoplasm without

16549

14

further characterization. The same report included a
a malignant neoplasm of tissue and 3
-plasmocytomas. No other study found a soft-tissue tumor.

In summary, the'Swedish studies of soft-tissue sarcomas
cannot be considered to have proved that exposure to phenoxy
herbicides is the cause of one or more types of this varied
group of malignant tumors. There are no fully reported
systematic studies to confirm what the Swedish investigators
describe as an association. There are an epidemiological
study gig) and observations of exposed populations that do
not support the finding as opposed to uncorrelated American
observations and an East German study ?19) that do strengthen

the case for such an association.

At best, the Scottish verdict of "Not proven" seems most
realistic at this time. The Advisory Panel on Toxic Sub-
stances of the American Medical Association says that "while
and pesticides (phenoxy herbicides in Agent
Orange) have been used in agriculture, forest management and
residential landscaping for over 30 years, there is still no
conclusive evidence that they and/or TCDD (a contaminant of
Agent Orange) are mutagenic, carcinogenic, or teratogenic in
man, nor that they have caused reproductive difficulties in

the human," 32:

3/5/83

L. B. HOBSON

1:



- 16550 

10.

11.

BIBLIOGRAPHY

Sundell, L., Rehn, M., and Axelson, 0. Exposure to?
herbicides - mortality and tumor incidence. An
epidemiological investigation in Swedish railway
workers. Lakartidningen 71:2466?2470. 1974

Axelson, O. and Sundell, L. Herbicide exposure,
mortality, and tumor incidence. An epidemiological
investigation on Swedish railroad workers. Work
Environ. Health 11:21-28. 1974

Axelson, O. and Sundell, L.
cancer. Lakartidningen 74:2887-88.

Phenoxy acids and
1977

Axelson, 0., Sundell, L., Andersson, K., Edling, C.,
Hogstedt, C., and Kling, H. Herbicide exposure and
tumor mortality. An updated epidemiological
investigation on Swedish railroad workers.
Work Envirn. Health 6:73-79. 1980

Scand. G.

Aspects on confounding in occupational
Scand. J. Work

Axelson, 0.
health epidemiology (Letter.)
Environ. Health 4:85-89. 1978

Views on criticism of pesticide studies
Lakartidningen

Axelson, 0.
(Reply to note by Jannerfeldt, E.).
1980

Axelson, O. A note on observational bias in
case-referent studies in occupational health
epidemiology. Scand. J. Work Environ. Health
6:80-82. 1980

59903?2 H00

Axelsdn, O. Testimony before EPA Hearing, EPA
Exhibit No. 587. 1980

Epidemiologic methodology and
1096. 1980

Jannerfeldt, E.
pesticide studies. Lakartidningen 77(12):

Remington, R. D. Specific summary critique of five
investigations related to concerns about Agent
Orange. Congressional Record, pp. 5-10911, 10912,
August 6, 1980.

Verbal testimony at EPA Hearing.
1980.

Hardell, L.
September 29,

16551

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

Hardell, L. Malignant tumors and
exposure to phenoxy acids - A clinical observation.
Lakartidningen 1977 

Hardell, L. and A. Malignant 
soft tissue tumors and exposure to phenoxy acids or
chloramphenacol. Lakartidningen 75:3535?3536. 1978

Hardell, L. and A. Case?control study:
Soft tissue sarcomas and exposure to phenoxyacetic
acids or chlorophenols. Brit. G. Cancer 39:711. 1979

Hardell, L. Relation of soft?tissue sarcoma,
malignant and colon cancer to phenoxy acids,
chlorophenols and other agents. Scand. G. Work
Environ. Health 1981

Axelson, O. Aspects of confounding and effect
modification in the assessment of occupational cancer
risk. J. Toxicol. Environ. Health 
1980

Eriksson, M., Hardell, L., Berg, N. 0., Holler, T.
and Alexson, O. Case-control study of malignant
tumors of the soft tissue and exposure to
chemical substances. Lakartidningen 76:3872-3875.
1979

Eriksson, M., Hardell, L., Berg, N. 0., Moller, T.,
and Axelson, O. Soft?:issue sarcomas and exposure to
chemical substances: a case?referant study. Brit. J.
Industr. Med. 38:27-33. 1981

9999212 Hog

Hardell, L. Malignant of histiocytic type
and exposure to phenoxyacetic acids or chlorophenols.
Lancet 1979

Hardell, L., Erikson, H., Lenner, P. Malignant
and exposure to chemical substances,
especially organic solvents, chlorophenols and
phenoxy acids. Lakaricningen 1980

Hardell, L., Brikson, H., Lenner P., and Lundgren, E.
Malignant and exposure to chemicals,
especially organic solvents, chlorophenols and
phenoxy acids: a case-control study. Brit. J.
43:169. 1981

Cancer

16552

22.

23.

24.

24a.

24b.

25.

25a.

25b.

26.

27.

28.

29.

Do phenoxy herbicides
1982

and Acheson, E. D.


Coggon, D.
cause cancer in man? Lancet

Hardell, L. and Axelson, O. Soft-tissue sarcoma,
malignant and exposure to phenoxy acids on
chlorophenols._Lancet 1982

Honchar, P. A. and Halperin, W. E. 

trichlorophenol, and soft-tissue sarcoma. Lancet
1981
Ott, M. G., Holder, 8. 8., and Olson, R. D. A

mortality analysis of employees engaged in the
manufacture of 2,4,S-trichlorophenoxyacetic acid.
J. Occup. Med. 22:47-50. 1980

Cook, kR. R., Townsend, J. C., Ott, M. G., and
Silverstein, L. G. Mortality experience of employees
exposed to 
(TCDD). J. Occup. Med. 22:530-532. 1980

and soft?tissue
1981

Cook, R. R. Dioxin, chloracne,
sarcoma. Lancet 

Zack, J. A. and Suskind, R. R. The mortality
experience of workers exposed to tetrachlorodibenzo?
dioxin in a trichlorophenol process accident.

J. Occup. Med. 22:11-44. 1980



A mortality study of workers employed at
Unpublished.

Zack, J. A.
the Monsant Plant in Nitro,
Soft?tissue sarcomas,

Moses, M. and Selikoff, I. J.

phenoxy herbicides, and chlorinated phenols. Lancet
1981

Johnson, P. 3., Kugler, M. A., and Brown, S. M. Soft
tissue sarcomas and chlorinated phenols. Lancet

1981

Hardell, L. and Erikson, M. Soft-tissue sarcomas,
phenoxy herbicides and chlorinated phenols. Lancet
1981

Sarma, P. R. and Jacobs, G. Thoracic soft-tissue
sarcoma in Vietnam veterans exposed to Agent Orange.
New Engl. J. Med. 1981

-165538

'30.

31.

32.

33.

34.

35.

36.

37.

38.

Barthel, E. Cancer risk in agricultural workers
exposed to pesticides. Arch. Geschwulstforsch.
1981

Bishop, C. M. and Jones, A. H. Non-Hodgkin's
of the scalp in workers exposed to dioxin.
Lancet 1981

Riikimaki, V., Asp, 8., and Hernberg, 8. Mortality
of 2,4-dichlorophenoxyacetic acid and 2,4,5-tri?
chlorophenoxyacetic acid herbicide applicators in
Finland. Scand. J. Work Environ. Health 8:37?42.
1982

Hogstedt, C. and Westerlund, B. Cohort study of
mortality rates of forestry workers with and without
exposure to phenoxy compounds. (No translation
available) Lakartidningen 1980

Smith, A. D. 0., Pearce, N. and Teague,
C. A. Do agricultural chemicals cause soft-tissue
sarcoma? Initial findings of a case-control study in
New Zealand. Unpublished report at ANZSEARCH Annual
Conference, May, 1982

Smith, A. 8., Fisher, D. 0., Giles, H. J., and
Pearce, N. The New Zealand soft-tissue sarcoma
case?control study: interview findings concerning
phenoxyacetic acid exposure. Unpublished report at
Third International Symposium on Chlorinated Dioxins
and Related Compounds. October 1982

Pattern of causes of
Hygiea

Edling, C. and Granstam, 8.
death among forestry workers, a pilot study.
1979

Tetrachlorodibenzodioxin: a survey of
Industr.

May, G.
subjects ten years after exposure. Brit. J.
Med. 39:128-135. 1982

Pazderova-Vejlupkova, J., Nemcova, N., Pickova, J.,
Lukas, 8., and Jirasek, L. The development and
prognosis of chronic intoxication by
tetrachlorodibenzo?dioxin in man. Arch. Environ.
Health 1981

8990212 HOD

18558

39.

40.

41.

42.

43.

44.

45.

46.

47.

TomatiET'L.

Cited in Huff, J. 8., Moore, J. A., Saracci, R. and
Long-term hazards of 
dibenzodioxins and dibenzofurans.
Environ. Health Prespectives 36:221-240. 1980; and

Hay, A. Accidents in trichlorcphenol plants: a need

for realistic surveys to ascertain risks to health. 
Ann. N.Y. Acad. Sci. 320:324. 1979

Thiess, A. H. and Frentzel-Beyze, R. Mortality study
of persons exposed to dioxin following an accident
which occurred in the BASF on Hovember 13, 1953.
Presented at the Fifth Interna:ional Conference on
Hedichem. September 1977

Thiess, A. M., Frentzel-Beyme, R., and Link, R.
Mortality study of persons ex seed to dioxins in a
trichlorophenol process accident that occurred in the
BASF AG on November 17, 1953. Am. J. Industr. Med.
3:179?189. 1982

Young, A. L., Calcagni, J. A., Thalken, C. E., and
Tremblay, J. W. The toxicology, environmental fate,
and human risk of Herbicide Orange and its associated

dioxin. USAF OEHL Technical Report (Report OEHL TR E:
78?92) pp. V-7, :2
Caramaschi, F., del Corna, G., Favaretti, C., PQ
Giambelluca, S. 8., Hontesarchio, 8., and Fara, G. M. 
Chloracne following environmental contamination by no
TCDD in Seveso, Italy. Intern. J. Epidemiol. C3
1981 

cn

o:

Hardell, L. Epidemiological studies on soft-tissue
sarcoma and malignant and their relation to
phenoxy acid or chlorophenol exposure. Umea
University Medical Dissertations, New Series No. 65.
1981

Anonymous. Phenoxy herbicides, trichlorophenols, and
soft-tissue sarcomas (Editorial). Lancet
1982

Phenoxy acids,
Lakartidningen

Hardell, L. and Axelson, O.
chlorophenols, and cancer.
1981

Council on Scientific Affairs' Advisory Panel on
Toxic Substances (American Medical Association). The
health effects of "Agent Orange' and 
dioxin contaminants. October 1, 1981

8 16355

2614-

-1-

AND HUMAN HEALTH

During the past decade, considerable controversy has developed over the
human health effects of exposure to TCDD, an unintentional contaminant
from the production of some herbicides and antibacterial agents and a
substance created by the process of combustion.

This controversy has been fueled by reports on the effects of Agent Orange
exposure on returning Vietnam veterans, the Love Canal situation, the
explosion of a 2,4,5-trichlorphenol plant in Seveso, Italy, and, most
recently, the buy-out by the U.S. government of Times Beach, Missouri, a
town contaminated with dioxin, 


During this time period, much speculation and misinformation regarding
this issue have been generated, with resulting confusion and apprehen-
sion in the public mind. It is the purpose of this paper to state the
facts regarding TCDD and human health and to place the issue in its
proper perspective.

This paper will document the following key points: 

C3

Certainly is a potent and toxic compound that can i?C
cause a variety of ill health effects at relatively low doses

in laboratory animals. We should be concerned with reducing 

any unreasonable risk of human TCDD exposure in our environ- 

ment. Based on the preponderance of scientific studies, the 

trace environmental presence of TCDD does not pose a health co

hazard to people. - a:

FQ

There is a wealth of epidemiological data supporting the
position that humans are far less susceptible to TCDD than
several of the animal species which have been studied.

The mere presence of TCDD in the environment (soil, water,
fish, etc.) is not proof of contamination from industrial
chemical sources.

- The scientific evidence does not demonstrate that TCDD
exposure causes cancer in humans.

There is no valid epidemiologic evidence linking TCDD exposure
in humans with a higher-than-normal rate of birth defects.

 



16556

-la-

TABLE OF CONTENTS

    0 I I I I I 
The American Medical Association Report . .
 Epidemiological Studies . . . . . . . . . .

Soft Sarcoma . . . . . . . . . . . . .

The Dow Mortality Study . . . . . . . . . .

Birth Defects . . . . . . . . . . . . . . . . .

The Toxicity of TCDD in Animal Species. . .
Environmental Hazard Evaluation (Soil/Fish)

ReferencesAUG

EBBEIIZ

1655?,



-2-

What is Dioxin?

The tenn "dioxins" (or more scientifically, "chlorinated dibenzo-p-

dioxins?) refers to 75 similar, but different substances.1

Each of the 75 dioxins that has been studied has its own unique identity

and toxicological properties.

0n the basis of animal studies, 
(TCDD) is considered to be the most toxic of the dioxins, and it has
been comprehensively examined for its toxicological effects. This

report will generally address the health impact of TCDD exposure.

TCDD is a highly toxic compound that can cause a variety of ill health

effects at relatively low doses in laboratory animals. We must be

concerned with establishing safe levels of human TCDD exposure in our

environment.

TCDD is formed as a trace unwanted contaminant in the production of

2,4,5-trichlorophenol from tetrachlorobenzene. The herbicide 2,4,5-

trichlorOphenoxyacetic acid other trichlorophenoxyacids, and
the germicide hexachlorOphene are from 2,4,5-trichlorophenol

and may contain trace amounts of TCDD and other dioxins.2

Dow has not produced the herbicide in the United States since

1979 due to reduced market demand from an Environmental Protection

usesnnz Koo

16558

Agency (EPA) emergency suspension which restricted some applications of
the product. There are no manufacturers currently producing in
the United States. A Dow joint venture company produces in New
Zealand. Dow currently supplies from inventories for the rice
and rangeland applications presently approved by EPA. The trace amount
of TCDD present in Dow-inventoried is less than 10 parts per

billion.

TCDD can also be created by combustion. Data indicate that chlorinated
dioxins may arise from the combustion of most types of organic material,
and suggest that chlorinated dioxins result from trace chemical reac-

tions occurring in fire.3

Many chemical reactions occur during combustion at very low concentra?
tions -- parts per billion and lower. With more advanced analytical
capabilities, scientists have discovered numerous sources where various

dioxins have been formed, including TCDD.

SB8EIIZ K00

Some of these sources include refuse incinerators, wood?burning stoves,
gasoline- and diesel-powered automobiles and trucks, fireplaces, char-
coal grills and cigarettes. TCDD and the other dioxin compounds have
been entering the atmosphere via airborne particulates from these

various sources and settling on soil and in bodies of water.

There have been a number of industrial incidents throughout the world

where people have been exposed to TCDD. Epidemiological studies have

16559-

-4-

evaluated these incidents and all of the evidence so far tends to reduce
the initial concern about the health effects of TCDD on humans. An
overall look at the mounting data has been undertaken by the American
Medical Association. These findings and conclusions are of interest and

bear mention.

The American Medical Association Report

 

In 1981 the American Medical Association (AMA) published a technical
report reviewing the medical evidence regarding the toxicity and long-

term health effects of the TCDD contaminant in Agent Orange*.



(D

The report's preface stated: 


"In spite of the voluminous is still very little Lu
Substantive evidence for many of the alleged claims that have co

been made against these compounds (dioxins). The most serious as

of these allegations assert that Agent Orange, or compounds of cm

a like nature, have caused malignant tumors, spontaneous abor-
tions and birth defects. Although data from studies on experi-
mental animals tend to support some of these claims, it is not
certain that the animal data are extrapolatable to man. No
laboratory animal can fully substitute for man; we must,
therefore, depend on the results of ongoing epidemiological
studies on persons who are known to have been exposed."

 

*(Agent Orange was a 50/50 herbicide mixture of and pro-
duced according to Specifications set by the U.S. government for use as
a defoliant in Vietnam to protect American troops from enemy ambush.)

16580

-5-

A close look at the summary of the report gives a clearer insight into

what the AMA's Advisory Panel on Toxic Substances found.

The AMA Panel reported that, in addition to the marked variations in the
sensitivity and susceptibility of animal species to all toxic substances,
there are significant differences between some of the toxic effects of
TCDD in experimental animals and the human experience. People appear to
be less sensitive to the health effects of TCDD than some of the animal

species studied.

One of the more pronounced biological effects of heavy exposure to TCDD,

as well as a number of other chlorinated aromatic compounds, is a tendency

to cause chloracne in certain animals and humans, the AMA Panel reported. Egg
This particular skin condition is regarded as the clinical marker for hd
TCDD over-exposure in people. The Panel noted that human systemic dis- 
orders from exposures to TCDD are unlikely to occur in the absence of z:
chloracne. Such findings as impaired liver and kidney function, gastro- ff:

intestinal irritation, muscle and nerve disorders, and depression and
irritation of the central nervous system have been reported after expo-
sure to relatively large amounts of TCDD. However, these disorders have
not been progressive, and they have disappeared with time, the AMA
report stated. In short, if there is no chloracne, there are unlikely

to be other persistent toxic effects from exposure to TCDD.

The AMA Panel also reported that other toxic effects of TCDD, when
experimentally produced in test animals, appear as pathological changes

in the liver, and epithelial tissue.

18581

-5-

TCDD is a powerful enzyme inducer. In addition to altering normal
enzyme activity, it may potentiate or inhibit the action of other

toxins.

The AMA report stated that TCDD may act as a promoter of carcinogenesis
in some strains of rats and mice. For cancer to start, it must go
through several distinct steps. The first stage, called initiation, is
produced by a carcinogen (an initiator) and probably involves irrevers-
ible mutational changes. Subsequent growth and development of an actual
cancer may require promotion, which may be the result of additional

applications of a carcinogen, or of other, non-carcinogenic materials.

CD

Promotion may involve additional unknown processes which could aid the C3
growth of the cancer or reduce resistance to the developing cancer.



TCDD is among the group of chemicals that may secondarily influence the formation of cancer by promotion. The carCinogeniCity associated with CD

TCDD in laboratory animals is typically accompanied by other signs of
systemic toxicity in contrast to some other chemical carcinogens for

which cancer is the only observable toxic effect.

TCDD has reportedly induced genetic changes in some forms of bacteria,
but evaluations in rodents and humans do not indicate that these

mutational changes will occur in higher animals, let alone people.

The AMA Panel made clear in its summary that the extensive information

collected for more than 30 years provides no conclusive evidence that

1658.2



and/or TCDD are mutagenic, carcinogenic or teratogenic in

humans, nor that they have caused reproductive difficulties in people.

Furthermore, the AMA report stated that undergoes relatively
rapid decomposition in the soil. However, TCDD does persist in soil

longer than but, in general, the half-life of TCDD in soil may

be no longer than one year. TCDD exposed to ultraviolet light is broken
down rapidly when present as a thin film on plants, water and the

surface of soil.4

Seveso

The most well?known and widely studied incident of community exposure to
TCDD followed the July 10, 1976, explosion at the ICMESA trichlorophenol
plant at Seveso, Italy. As a reSUlt of that accident, approximately
seven square miles of the were contaminated by chemicals,

including TCDD, exposing as many as 30,000 people.5?6 The heaviest con-

Beasllz'xoa

tamination involved an area of 180 acres (Zone A), occupied by 736 people.

TCDD was reportedly detected on soil at levels in excess of 5.5 parts per

million (ppm).7a5 In general, little effect was noted on plant life, but

wild animals did die, particularly in Zone A (nearest the chemical plant).

About four percent of the domestic animals in contaminated zones died,

and virtually all of these were small animals.5 Some of these deaths

may have been due to other chemicals released at the same time. ..

10563

-8-

Chloracne, which has generally been regarded as the hallmark or sentinel
sign of TCDD exposures, was observed in this population, although at
generally low rates and primarily in children.9:10s5a5 The highest
incidence of chloracne reported was about 13-14 percent among elementary
school children in Seveso, and this was generally mild and rapidly
resolved in most affected people.5=5a11 Other than chloracne (of which
there were ultimately 100-200 cases), nausea, vomiting, itching and head-
ache were the most commonly observed readily attributable to
the exposure which may have involved chemicals other than TCDD. Head-
ache, stomach and intestinal upset were more commonly noted in

individuals with chloracne.9a6

CD
Bisanti12 concluded that no "major event? had taken place with respect ii:
to birth defects, miscarriages (spontaneous abortions), births and no
deaths. While this seems to be the consensus, changes in birth defect gig
reporting and in the reporting of "spontaneous" (sometimes "thera- 
:3

peutic") abortions have obscured any small changes which might have
occurred.13:5.5.14:11 Data on reproductive outcome are limited in
number, but Reggiani11 reports that examination of fetuses in 34 cases
of medically-induced abortion revealed no evidence of injury attribu-
table to TCDD. Pregnancy outcome in the years following 1976 appears to

be comparable with Western experience.5

Of particular interest is the fact that no birth defects were observed
0

during 1977 among the 70 births in Zones A and 8. Only those pregnan-

cies coming to term after January 1977 would have been relevant, 

165364

-9-

those in which July 1976 maternal exposure would have taken place during

the critical period for the developing fetus.

Many factors are known to cause birth defects in humans, which occur in
three to six percent of live births: viral infections, genetic predis?

position, smoking, alcohol, and even medications, including excess

quantities of certain vitamins.

Epidemiological Studies

In recent years a number of investigations have been conducted concern-

ing the human health effects associated with exposures to TCDD.

The following brief review of the human exposure experience with TCDD
and the epidemiological studies of those pepple are taken from a recent
report by the Advisory Committee on Pesticides, which is charged with

making recommendations concerning the safety and safe use of pesticides

in the United Kingdom.

121 workers were

- Monsanto's plant at Nitro, West Virginia:
No apparent

followed up for 30 years after exposure to TCDD.

excess in total mortality or in deaths from cancer or diseases

of the circulatory system was observed.

- Coalite plant (1968) in the United Kingdom: Chloracne was 
initially seen in 79 workers and some had initial abnormali-
ties in the liver but these returned to normal limits within
10 days. No clinically recognizable disease has been demon-
strated. There is no evidence of liver or cardiovascular

disease.1

?00



18535

-10..

- Dow Chemical Company workers exposed to No adverse

effects have been noted in an epidemiological study in these
workers (mortality was actually less than expected).1

Vietnam: Exposure to Agent Orange is blamed for a host of
illnesses. The U.S. Veterans Administration found that no
cause-effect relationship was established between TCDD
exposure and alleged medical complications.15

Finnish railroad workers using and other phenoxyacetic
acid herbicides in forestry and on roads and railways: 1,960
men were Sf?died_an??no increased mortality from cancer was
demonstrated. There were no deaths from cancer in forestry
workers although six deaths would have been statistically

predicted.15

Soft Tissue Sarcoma

The most persistent hypothesis deals with the possible causal associa-

tion between TCDD and soft tissue sarcomas. This hypothesis is derived 3

from the epidemiological work done by Dr. Hardell and his associates in 

no

Sweden.16 

(A)

cc:
Soft tissue sarcoma is a generic tenn for a group of lesions that 

rs:

include more than 100 different types of tumors.17 These tumors are
relatively rare; thus few pathologists have been able to achieve a high
degree of consistency in their diagnosis.18 Most experts believe that
if TCDD were acting as a carcinogen it would cause an increase in one I
type of soft tissue sarcoma, but would be unlikely to raise the risk all
across the board. One of the reasons vinyl chloride is accepted as a.
human carcinogen is the specific way it acts. At high levels, it

produces a single type of cancer: angiosarcoma of the liver.

18566_

-11-

The question which arises is: If there is a common exposure link
between various reports of soft tissue sarcomas in occupational groups,

is TCDD the link?

The exposure documentation differs from study to study. In some, based
on detailed technical knowledge of the chemical process, analyses of
reaction contaminants, and clinical evidence of excessive group exposure
'(such as chloracne), there is evidence that TCDD is at least one of the

common exposure links. In other reports, the evidence is more presump-

tive, and in some cases it is totally lacking.

Epidemiologists from Dow Chemical and Monsanto have reported on the soft


tissue sarcoma cases occurring in their occupational groups presumably 
exposed to high levels of 

I ax:

In 1964, a group of Dow trichlorophenol production workers experienced (L:
c:

Cx)

an outbreak of chloracne. Sixty-one workers were identified as being
involved with the trichlorophenol process. Forty-nine of these workers
developed a rash diagnosed as chloracne. Epidemiologists followed up on
the status of all 61 men through 1978, 14 years after the incident.
Statistically, 7.8 deaths would have been expected in this group, but
only four had occurred. Among the causes of death, one was from cardio-
vascular disease when 3.8 would have been expected. Cancer was idehti;
fied as the cause of three deaths when 1.6 were expected. None of these
findings represent a statistically significant deviation from the

expected experience.20 One of the cancer deaths in the Dow group was

1658?

-12..

due to a type of soft tissue sarcoma called fibrosarcoma. In 1981
another employee in the Dow trichlorophenol worker group developed
another type of soft tissue sarcoma, malignant fibrous histiocytoma, one

of a multitude of tumors which comprise the soft tissue sarcoma category.

The employee died in 1983.

In the 1978 Monsanto study, no link between exposure to TCDD and excess
deaths due to cancer or other causes was found among 121 current and
fonmer workers employed at a plant in Nitro, West Virginia. The employees
who participated in the study included workers who developed chloracne

as a result of over-exposure to TCDD in a manufacturing accident in 1949.

Among the Monsanto workers studied, 32 deaths were reported versus 46
which would be expected. Cancer accounted for nine deaths among the
Monsanto group compared with more than nine deaths expected.

One of these cancer deaths was attributed to soft tissue sarcoma, a

malignant fibrous histiocytoma.

11068112 #500

A second Monsanto study evaluated the mortality experience of male

hourly workers employed at the Nitro plant during or after 1955 and
followed through 1977.22 When adjustments were made for prior exposures
to a known bladder carcinogen, the observed deaths due to all causes and
due to cancer were less than expected. Among the subset of employees I
who had worked in production, the preportion of deaths due to
cancer was lower than that found in the rest of the plant population.

In this subset one individual died as a result of a soft tissue sarcoma,

a liposarcoma. 153538

-13..

These four soft tissue sarcoma cases were summarized in the literature
in 1981.23:24 All four had occupational exposures to trichlorophenol.
Only one, the second Monsanto case, had also been exposed to 
Three had frank chloracne and the fourth, a Dow employee, a facial rash
during his assignment in the trichlorophenol plant. All four had a
history of cigarette smoking. It was suggested that the hypothesis of
soft tissue sarcomas being caused by heavy TCDD exposure among cigarette

smokers needed to be tested.

Three other cases of soft tissue sarcoma were also reported in 1981.25:25
Exposure to trichlorophenol or TCDD was not well documented.

In one case, the individual was considered unexposed until his disease
The other two occurred in a father and son.

was discovered.

In a group of Dow employees (none of whom had chloracne)27 there

have been no cases of soft tissue sarcoma.

At Dow, the mortality experience of 204 persons exposed to 

during its manufacture from 1950 to 1971 was studied through 1978. No
adverse effects were observed with respect to occupational exposure to
either or its feedstock, 2,4,5-trichlorophenol. Mortality of

the studied workers was favorable compared to that of U.S. white males.

There have been no cases of soft tissue sarcoma reported in a DASF

worker group, many of whom experienced severe chloracne. Seventy-four

HUG

SDSEIIZ

-14-

members of the BASF group were exposed to TCDD during a 1953 accident in
a trichlorophenol production unit or during cleanup and repair following
the accident. Exposures were heavy. Twenty-seven years after the
accident a mortality study of those exposed to TCDD in the accident was
undertaken. Overall mortality did not differ in this group from the
rate expected in three external reference populations or from that

observed in two internal comparison groups.28

Dow is cooperating with the National Institute for Occupational Safety
and Health (NIOSH) to prepare a study of more than 4,000 industrial

workers potentially exposed to dioxins.

 

CD
CD

Swedish Epidemiological Studies 
ex:
Dr. Hardell's work, which has lead to the question about soft tissue cg:
c:
sarcomas, uses a different methodology than in the foregoing studies -- CD

the case control approach -- and reports on different patterns and levels
of exposure.15s29 In his first study, he reported a five- to six-fold
increased risk of soft tissue sarcomas for those presumably exposed to
phenoxyacetic acids, predominantly and chlorophenols (?pre-
sumably" because the exposure information was subjectively reported by
the study participants and/or next of kin). Objective records were
incomplete and were difficult to interpret. In the second study, he
also estimated the risk associated with exposure to phenoxyacetic acids

not contaminated with TCDD and found the same magnitude of risk.

-15-

Some epidemiologists reviewing the information find this confusing. The
justification for combining chlorophenols and was the presumption
of a common contaminant, TCDD.39 If that were the case, increased risk

would not be expected for the phenoxyacetic acids without this contaminant.

In both of his studies, Dr. Hardell first administered mail questionnaires
and then followed up with telephone interviews on a selected subset of
subjects. While the questionnaire was designed with the idea of masking
the intent of the investigation, the telephone inquiry Specifically

focused on the exposures of pertinent interest: phenoxyacetic acids and

chlorophenols.

Interestingly, in his second study, Dr. Hardell reported the frequency

non

of the various exposures among both cases and controls and most of the
risk estimates were elevated. The risk from only one out of 15 chemi-

cals, sodium chlorate, was not elevated. Nicotine had a high risk.

[058112

Unfortunately, the risk estimates derived from the questionnaire alone
are not known, nor is it known what the risk estimates would have been.
for other agents, if they had been probed in the same depth as were the

phenoxyacetic acids and chlorophenols.

There are a number of other concerns relating to Hardell's work. The

accuracy with which exposure to the chemicals is determined is of vital
importance, and the memory recall techniques Dr. Hardell used are subject

to error. Workers are unlikely to remember with accuracy the chemicals

they used some years in the past. It is also extremely difficult to  

estimate the extent or duration of the exposures. Furthermore, the

-15-

identification and classification of the various soft tissue sarcomas
have not been consistent. The diversity of tumors is particularly
difficult to explain. If all the tumors had been of a single type, the

data would be much more convincing.

Furthermore, research currently underway does not support Hardell's

hypothesis concerning soft tissue sarcoma:

- The U.K. Government Advisory Committee on Pesticides noted in
February 1983: "It is our view that the procedures used for
establishing the Swedish study groups, and some aSpects of the
collection of the exposure data, were not reliable."

Smith and associates30 are conducting a case-control study in New

Zealand. Herbicide application is a registered profession in New 
Zealand, and phenoxyacetic acids have been used in bulk since the 53
late 19405. In a preliminary report presented at the 1982 Inter- ?45
national Symposium on Chlorinated Dioxins and Related Compounds,
Smith noted he was unable to establish an association between the :3
use of phenoxyacetic acids and soft tissue sarcoma. In fact, not 
one of the soft tissue sarcoma patients had ever worked as a (J)
licensed phenoxyacetic acid herbicide applicator. a:
- Milham31 explored this issue in the state of Washington using death 

certificates. While there has been considerable exposure over the
years to phenoxyacetic acids and chlorophenols in Washington state,
occupations related to their use did not show consistent patterns
of death due to soft tissue sarcomas. Surprisingly, the two occu-
pations with the highest proportional mortality ratios were marine
engineers and bankers.

- hnong the deaths in the U.S. Air Force "Ranch Hand" Study, none
were due to soft tissue sarcomas.32

Riihimaki studied 1,960 Finnish herbicide applicators and found no

difference in death rate from any natural cause, including cancer,
conpared with the total Finnish male population. There were no 

soft tissue sarcomas reported in this group of workers.3

F.

- The Michigan Department of Public Health (MDPH) is investigating
the possible relationship between dioxin and a small number of soft
and connective tissue cancer deaths among women in Midland County

(where Dow has facilities).34 In a recent report (May 1983) MDPH

.16572

-17..

did not identify a specific association between any environmental
factor and the reported increase of soft tissue cancer in the
county. While overall age-adjusted cancer mortality rates in
Midland County were below the state of Michigan average, soft and
connective tissue cancers among white females in the county were
elevated. As part of the initial study, MDPH reviewed data for 28

other U.S. counties in which TCDD or other dioxins were likely
produced as a chemical manufacturing contaminant. They found no

increase of soft tissue cancer in these counties verSus counties
that did not have industrial manufacturing sources suspected of

generating such dioxins.

Various animal species did not develop soft tissue sarcomas when
fed TCDD during laboratory experiments.

Additional research is under way in the United States and elsewhere.
The results of these studies should offer a better perspective on this
issue. But right now, the preponderance of evidence does not demon-

strate a link between TCDD exposure and soft tissue sarcomas.

The Dow Mortality Study

6068112 tog

In 1976 a research study was published that summarized the mortality
experience of over 8,000 men employed by The Dow Chemical Company. The
population of interest was defined based on a March 1, 1954, employee

census of the Midland, Michigan, manufacturing location of the Company.35

The study originally developed background mortality data on the Dow
worker population so that findings obtained from smaller groups directed
at more specific questions, questions regarding particular chemi-
cal exposures or processes), could be placed in perspective. The 

observation period for mortality follow-up was 1954 through 1972, and

"18573

-18-

comparisons were made to the mortality experience of the corresponding
United States white male population. At that time, mortality among Dow

employees was found to compare favorably with that of the general U.S.

population.

The objectives of the most recent update of this group of 8,181
employees were to extend the observation period through 1978, and to
include comparisons with a community-based group of employed men not
engaged in chemical manufacturing activities. In addition, the effects

of cigarette smoking on mortality were investigated.

The Dow group consisted of personnel from research departments, the
corporate headquarters, and a major manufacturing division of the

Company. It has been estimated that as many as 500 distinct chemical

woo

processes ranging from small batch operations to large continuous and
highly-automated production units have been onstream at the Midland

manufacturing site, including production of trichlorophenol and related

products.36

For the most part, the men in the study represent long-term Company

employees. Some were hired prior to the 19405.

Cause of death was obtained fran death certificates for all but nine of

the 1,932 deceased members of the study population. .

118574,

-19-

Overall mortality in the Dow group was 19 percent less than that
expected for the corresponding United States population. Expressed in
another way, a Dow worker of age 25 in 1972 could expect to live

2.2 years longer than a non-Dow worker of the same age.

There were no statistical findings of excess mortality among Dow
employees for any cause of death category. For total malignant
neoplasms, there were five percent fewer deaths observed than expected
based on the United States white male p0pulation. Significantly fewer
deaths were observed than were expected for all categories. Presumably,
if low-level TCDD exposure caused a variety of increased illnesses, the

opposite from what is stated above would have been found.

Comparing the Dow workers with a comparable employee population in
Michigan, the overall mortality risk was about 10 percent lower in the

Dow group than the corresponding non-Dow group.37

IIBEHZ M00

Birth Defects

Concerns that TCDD may cause birth defects are not substantiated by
epidemiological science. A brief review of some areas of concern and

the corresponding results, and a more specific look at the Dow situation'

in Midland, Michigan, are presented:

a Aerial Spraying of in Oregon: Claims of a link
between incidence of miscarriages and herbicide could
not be established when all the data were available.15

-20-

A six-year study of agriculture and forestry workers in
Hungary exposed to Data do not suggest that there
has been any significant increase of birth defects.1

New Zealand workers exposed to An alleged abnormally
high incidence of neuraT tobe defects (such as spina bifida)

were chance occurrences, and there is "a very high assurance
of safety in normal use" of 

- Australian worker exposure to and Claims of
high incidence of birth defects were investigated, and
available information revealed no evidence that the birth
defects were caused by exposure to these two compounds.15

Facial cleft defects in Arkansas: Study of exposure to
from 1948-74 reveals these defects to have no
association with 

About five years ago it was reported that the birth defect rates in
Midland County (Michigan) were three times above those for the state of
Michigan. At that time, the Michigan Department of Public Health (MDPH)
examined this issue. It found an increase had occurred between 1971 and

1974 due to additional reporting of minor congenital malformations.

ZIEEHZ M00

Before 1971 and after 1974, the birth defect rates in Midland County
were either at or below state rates. If wideSpread chemical contamina-
tion of the environment was responsible for birth defects, the rates
should have gone up and stayed up. They did not. The changes may have

been due to reporting techniques, not additional cases of birth defects.

It is very important to note that the numbers of county births and birth
defects were small, and thus, rates calculated from them tended to be

more variable from year to year than the comparable rates for the entire
state. The MDPH recently announced (May 1983) an updated report which I
concludes that the birth defect elevations noted in the early 19705 were

not unique and that no additional studies are warranted at this time.38 



In 1982 Dow scientists published a report of a survey of reproductive
events among 370 wives of Dow Michigan Division employees exposed to
dioxins.39 They analyzed the associations between exposure and spon-
taneous abortion (miscarriage), stillbirths, infant deaths, and several
categories of congenital malformations before and after controlling for
multiple confounders sources of error or confusion) independently
and in various combinations. They did trend analysis to determine if
duration of paternal exposure was related to adverse pregnancy outcome.
There were no associations between any exposure and adverse pregnancy
outcomes. These findings have recently been confirmed in another study
conducted by Australian scientists40 who examined the impact on birth

rates involving Vietnam veterans potentially exposed to Agent Orange.

The Toxicity of TCDD in Animal Species

There are wide variations in how different animal species are affected

man

by TCDD. For example, the single oral dose L050 (the amount that it
would take to kill 50 percent of the test animals) in guinea pigs is
0.6-2.0 ug/kg (micrograms per kilogram or every 2.2 pounds of body
weight [a microgram is one millionth of a gram; a kilogram is 1,000
grans, or 2.2 In hamsters, however, TCDD is much less

toxic, with an oral L050 of 1157-5051 

Therefore, the guinea pig is approximately 5,000 times more sensitive
than the hamster, although both species are closely related biologi-

cally. The direct relevance of any single animal experiment to humans

must be evaluated with respect to all other available data.

16577

-22-

In acute and subchronic studies, liver toxicity is a prominent component
of TCDD toxicity in rats, mice and rabbits, but not in monkeys, where

effects on the bone marrow and epithelial tissue are more prominent.

Thymic atrophy in early toxicity studies suggested that TCDD might
decrease the immune response. In laboratory animals, scientists can
induce immunotoxic effects with high doses of TCDD. However, as the

Seveso incident shows, these effects have not been demonstrated in

exposed humans.45.45

Teratogenicity/Animal Species

0f the various toxic responses of animals to TCDD, the potential for

nag

teratogenicity (birth defects) has perhaps received the most attention.
Teratogenic effects resulting from TCDD have been realized only in mice,

which show an increased frequency of cleft palate, along with an abnor?

mality of the central collection system of the kidney. It is well known
that many factors can cause birth defects in mice, including the stress
of being transported by air during pregnancy, or being deprived of
drinking water overnight. In both rats and mice, dose levels of TCDD

have been identified at which these effects do not occur.

In rats, TCDD does not cause a teratogenic effect but sufficiently high?

doses can cause embryo- and fetotoxicity.

18578

-23..

Many of the studies with TCDD in monkeys have been conducted at the
University of Wisconsin. In experiments where the high dose level of
about 0.011 ug/kg/day TCDD in the diet was given to the monkeys for up
to 9.3 months, there were substantial toxic effects.47,48 A preliminary
abstract of a follow-up study49 showed that monkeys given diets contain-
ing approximately 0.0017 ug/kg/day of TCDD had only slight toxicity.

There are studies currently in progress at the University of Wisconsin

at lower doses.

In a three-generation reproduction study50 of rats at incremental dose
levels of TCDD in the diet, high doses caused decreased fertility and
neonatal survival. The intermediate dose level caused decreased fertil-
ity and other effects in the first two generations, but not in the
third. At the low?dose level, there was no impairment of reproductive

capacity through the three consecutive generations, indicating that

SIBEHZ 3mg

0.001 ug/kg/day was a no-adverse?effect level over multiple generations.

Mutagenicity/Animal Species

A number of mutagenic studies have been conducted with TCDD. The major-
ity of the tests have been negative or uninterpretable. Only two of the

tests were positive in one strain of Salmonella bacteria, showing that

there is low possibility for mutagenesis with TCDD in bacteria.

In studies45,51:52:53:54,55,55,57 with rats or human cells, there is

little indication that TCDD elicits a mutagenic response.

165? 9.

-24-

Thus, while a few positive or questionable mutagenic responses have been
observed in certain plant or microbial test systems, there appear to be

no definitive correlates of mutagenicity in higher animals or humans.
Carcinogenicity/Animal Species

Carcinogenic studies following ingestion of TCDD have been conducted in
rats and mice.58a59:50 Review of these data indicates good correlation
of the results in rats and mice, with a carcinogenic response associated

only with lifetime ingestion of higher dose levels that also induce

other toxicity. The liver was the primary target for cancer in both E:
rats and mice. No cancer response occurred at dose levels of 0.001- 
0.0014 ug/kg/day in the rats and 0.001-0.03 Ug/kg/day in mice. ?2
cxa

cc:

TCDD does cause cancer in animals. However, it is only at very high 


dose levels dose levels that are higher than those that elicit other
kinds of toxicity. Cancer induced by exposure to TCDD would be preceded
by substantial signs of toxicity. This toxicity (which is reversible
when exposure st0ps) would act as a sentinel, or warning at dose levels

below those that might cause cancer.

Environmental Hazard Evaluation

Soil

 

Potential human exposure to TCDD from environmental contamination
concerns many people. The following factors should be considered when

evaluating potential human health hazards due to TCDD in soil: 

-25-

TCDD is tenaciously bound by soils and other materials (carbon,
charcoal), and studies have shown that plants grown in soils

containing TCDD do not accumulate or translocate the material.

The amount of material absorbed depends on contact with the skin,

or ingestion.

Soil binding of TCDD reduces the amount that can be absorbed

through the skin or internally from ingestion.

Animal studies show that only a small amount of TCDD is absorbed
from contaminated soil. In animal studies, when such soil con-
tained about 500 parts per billion (ppb) TCDD and when the soil
was held in contact with skin for 24 hours (covered with aluminum

foil), about 1/1000 of the TCDD in soil was absorbed.

llBEllZ you

Estimates of absorption from the Seveso incident suggest that

people would be expected to absorb about 1/2000 of the average
amount of material present in 1 square meter (approximately

1.2 square yards) of their environment each day. If one uses the
laboratory or Seveso data, one can estimate that 100-1000 TCDD
in soil could result in human absorption of as much as a dose
equivalent to that which produced no effect when fed to laboratory
rats for a lifetime. If one assumes that soil is eaten in signifi?'

cant amounts by children, the permissible level is less, perhaps

10 ppb.

10581

-26..

Based on the assumptions and calculations presented here, (particularly
those based on the work of Dr. Kingsley Stevens),61 it appears that
general environmental soil contamination in the range of low parts per
billion would not pose a health hazard to the general population,
including the unborn, children, pregnant women or the old and infirm.
For the general population, excluding children with intimate soil con-
tact or those who ingest soil, a level of 100 would still pose no
hazard. In occupational settings, where workers are protected by
standard industrial hygiene practices, higher levels of contamination

would not be accompanied by greater exposure or hazard.

Dioxin and Fish

 

Rats consuming a diet containing 22 parts per trillion (ppt) TCDD

BIBEHZ H00

(1 nanogram-TCDD/kg of body weight per day) for a lifetime showed no
adverse health effects due to TCDD. Rats consume about 10-20 percent of
their body weight in food each day, while people daily consume about

2-3 percent of their body weight. Thus, for a given TCDD concentration,
rats will ingest more of the compound than people. A person eating
about 2-3 pounds of food per day uniformly contaminated with 25 TCDD
would ingest about 0.5 nanogram/kg body weight/day or about one-half the
dose (weight of TCDD per unit of body weight) shown to produce no
adverse health effects when fed to rats for a lifetime. Again, this is

true because people eat less food in proportion to their body weight

compared to laboratory rats.

18582

-27-

The dose received by eating contaminated fish is much less. Most people

in the United States (99 percent according to U.S. government figures)
eat less than 0.6 pound of fish per week. Nine out of ten people in the
United States eat less than 0.3 pound of fish per week, while the
"average" person eats still less. Since fish is such a small part of
the Anerican diet, 99 percent of the American public would receive less
than 1/70 of the amount of TCDD shown to produce no effect in laboratory
rats fed TCDD daily for their lifetimes, if those people ate fish con-
taining 25 of TCDD. 0r said another way, a person could eat nearly

one ton of fish per year containing 25 of TCDD and not exceed the

no-effect level established by laboratory animal experiments.

The fish consumption guideline of 25 established by the Food and

Drug Administration assures an adequate margin of safety to protect

people from overexposure to TCDD.52

5159112 H00

August/1983 (6)

?Wl658&

10.

11.

12.

13.

-28-

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-30-

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Tumors and Exposure to Chemical Substances.
Lakartidningen 76:3872-75, (EPA Translation).

Smith, A. H., et al., (1982): The New Zealand Soft Tissue Sarcoma
Case-Control Study: Interview Findings Concerning Phenoxyacetic
Acid Exposure, Third International Symposium on Chlorinated Dioxins
and Related Compounds, Salzburg, Austria, October 12-14, 1982.
Milham, 5., (1982): Lancet,
1:1464-1465, 1982.

Herbicides, Occupation and Cancer.

Status of the Ranch Hand Study Concerning

Agent Orange, Presentation to the House Committee on Veteran's
Affairs Subcommittee on Oversight and Investigations.

Riihimaki, V., et al., (1982): Mortality of 2,4-Dichlorophenoxy-
acetic Acid and 2,4,S-Trichlorophenoxyacetic Acid Herbicide
Applicators in Finland. Scand. J. Mark. Environ. Health, 8:37-42.

ZZBEIIZ H00

"Evaluation of Soft and Connective Tissue Cancer Mortality Rates for
Midland and Other Selected Michigan Counties Compared Nationally and
Statewide," Michigan Department of Public Health, May 4, 1983.

Ott, M. G., et al., (1976): Determinants of Mortality in an
Industrial Population. J. Occup. Med., 18:171-77.

Ott, M. G., et al., (1975): Linking Industrial Hygiene and Health
Records. Am. Ind. Hyg. Assoc. J., 36:760-766.

Ott, M. G., (1982): Effects of Selection and Confounding on
Mortality in an Occupational Cohort (Doctoral Dissertation)
(Mortality Among Men In A Chemical Manufacturing Company --
Executive Summary), University Microfilms, Ann Arbor (In Press).

?Evaluation of Congenital Malformation Rates for Midiand and Other-
Selected Michigan Counties Compared Nationally and Statewide",

1979-1981, Michigan Department of Public Health, May 4, 1983.

Survey of Reproductive Events of

Townsend, J. C., et al., (1982):
An. J. Epidem.,

Wives of Enployees Exposed to Chlorinated Dioxins.
115:695-713.

18586

40.

41.

42.
43.
44.

45.

46.
47.
48.
49.
50.
51.

52.
53.
54.

55.
56.

57.
58.
59.

-31-

Australia Reports No Link Between Service in
The Epidm. Monitor,

8., (1983):
Vietnam and Birth Defects Among 



Schwetz, B. A., et al., (1973): Environ. Health Perspect. Exp..
Issue No. 5:87.

McConnel, E. E., et al., (1978): TOxicol. Appl. Pharmacol., 44:335.
Olson, J. R., et al., (1980): Toxicol. Appl. Pharmacol., 55:67.
Henck, J. H., et al., (1981): Tbxicol. Appl. Pharmacol., 59:405.

Homberger, E., et al., (1979): The Seveso Accident: Its Nature,
Extent and Consequences, Report of the Givaudan Research Company.
Reggiani, G., (1980): J. Toxicol. Environ. Health, 6:27.
Allen, J. R., et al., (1977): Toxicol. Appl. Pharmacol., 41:177.
Allen, J. R., et al., (1977): Food Cosmet. Toxicol., 15:401.
Schantz, S. L., (1979): Toxicol. Appl. Pharmacol., 
Murray, F. J., et al., (1979): Toxicol. Appl. Pharmacol., 50:241.

Green, S., Moreland, F. S., (1975): Toxicol. Appl. Pharmacol.,

33:161.
Green, S., et al., (1977): DA By-Lines, 6:292.

82581 13 H00

Hassom, J. S., et al., (1977-78): Hut. Res., 47:141.
Khera, K. S., Ruddick, J. A., (1973): 
Perinatal Effects and the Dominant Lethal Test in Histar Rats?, in
Chlorodioxins - Origin and Fate. Advances in Chemistry Series, No.
120 (Etcyl H. Blair, American Chemistry Society, Washington, DC.
Beatty, P. H., et al., (1975): Toxicol. Appl. Pharmacol., 31:309.
Tenchini, M. L., et al., (1977): Approaches to Examination of Gene-
tic Damage After a Major Hazard in Chemical Industry: Preliminary
Cytogenic Findings on TCDD-Exposed Subjects After Seveso Accident,
Special Project of Investigations on TCDD?Exposed Pregnancies (Prof.
G. B. Candiani and Prof. L. DeCarli), University of Milan, Italy.

Rehder, H., et al., (1978): Schweiz. Med. 108:1517.
Kociba, R. J., et al., (1978): Toxicol. Appl. Pharmacol., 46:279.

National Cancer Institute, (1980): DHHS Public. No. NIH80-1765.

18587

60.
61.

62.

-32-

Toth, K., et al., (1979): Nature, 278:548.

Stevens, K. M., (1981): Agent Orange Toxicity: The Quantitative

Perspective. Human Toxicology. 1. PP. 31-39.

Cordle, F., (1981): The Use of Epidemiology in the Regulation of
Dioxins in the Food Supply. Regulatory TOxicology and Pharmacology

1, Academic Press, pp. 379-387.

August/1983 (6)

Hug



a:






Environmental
Infomamn


I



2815

For Further Information Contact:
R. N. (Bob) Charlton
(517) 636-9303

Health 

EBszuNh

AND HUMAN HEALTH

Executive Summary

The issue of dioxin in the environment and the possible human health
effects of this compound has been the subject of much Speculation over

the past decade.

This paper discusses scientific research and makes the following major
points:

The word "dioxin" refers to a family of 75 compounds of which the
one called is the most toxic based on animal tests. Dioxins
can be produced as an unwanted contaminant of some manufacturing
processes and through some combustion sources.

An American Medical Association review of available research has
concluded that there is no convincing support for allegations that
TCDD causes cancer, birth defects or other reproductive difficul-
ties in humans.

Studies of some 30,000 people exposed to TCDD after an industrial
accident in Seveso, Italy, found a chemically-induced acne that
occurred in approximately 200 people and no persistent effects
which could be verified. There was no excess of birth defects,
abortions or deaths in the population.

A number of studies of workers accidentally exposed to large

amounts of TCDD as long as 30 years ago have identified cases of
chloracne in some of those workers. However, no long-term health

effects have been demonstrated among this worker group. The
studies have looked for cancer, heart disease, liver damage and
other ailments.

OBBETTZ hood.

The work of a Swedish scientist who purports to find a link between
TCDD exposure and soft tissue sarcoma (a relatively rare cancer) is
not consistent with other studies. This research is not supported
by the bulk of studies of exposed industrial employees, herbicide

applicators and others which show no excess of this type of cancer.

A study of 8,000 Dow employees in Midland, Michigan, where herbi-
cides were produced, found their death rate to be 19 percent less-
than the general U.S. pepulation. There was five percent less
cancer than predicted.

 



1 18563-

THE DOW CHEMICAL COMPANY 0 MIDLAND, MICHIGAN 48640

Investigations of herbicide applicators, forest workers and indus-

trial workers and their wives found no link between TCDD exposure
and birth defects.

- Studies of animals exposed to TCDD show a variety of adverse health
effects that vary widely from species to species. In most studies,
safe levels of dioxin exposure have been identified. Cancer

occurred only at levels where animals were suffering from obvious
TCDD poisoning.

A review of the scientific literature suggests that humans are less
susceptible to TCDD than various animal species.

As is the case with other toxins, TCDD exposure should be carefully con-
trolled. The scientific evidence to date does not warrant undue concern
about individual exposure to trace amounts of TCDD presently measured in
the environment. Government environmental guidelines currently provide

for more than adequate margins of safety.

1400

1888112

ii

185301

Analytical Studies
for the US.
Environmental Protection Agency

VOLUME Ila
Decision Making
in the Environmental

Protection Agency 
CASE 

.ZUSllt?lUleOU

Prepared by

Robert E. Burt, Nathan J. Karch,

Raphael G. Kasper, Richard W. Lowerre,
Lawrence E. McCray, and J. Roxanne Arno/d
for the

Committee on Environmental Decision Making

Commission on Natural Resources
National Research Council

 

NATIONAL ACADEMY OF SCIENCES .
Washington, D.C. 1977 - .  

165291

 



EXPLICIT CRITERIA AND PRINCIPLES FOR IDENTIFYING CARCINOGENS:
A FOCUS OF CONTROVERSY AT THE ENVIRONMENTAL PROTECTION AGENCY

Nathan J. Karchl

CONTENTS
I. DESCRIPTION OF THE DECISIONS
II. INTRODUCTION
A. The Nature of the Disease
B. Cost and Extent of Cancer
C. Environmental Origins of Cancer-
D. Strategy for Control: New Emphasis on





.

Prevention

CANCER PRINCIPLES

Identification of Carcinogenic Substances

Early Development of Principles

Statutory Framework

Use of Cancer Principles in the DDT
Cancellation Proceeding

Use of Cancer Principles in the Aldrin/Dieldrin
Suspension Proceeding

Use of Cancer Principles in the Heptachlor/
Chlordane Cancellation and Suspension
Proceedings

1. Magnitude of the Cancer Problem

2. Uncertainty in the Mechanism of Cancer
Induction 

3. Establishment of Carcinogenicity and
Assessment of Risk .

4. Problems with Experiments in Animals

Use of Cancer Principles in the Mirex Fact?
Finding Proceeding

Use of Cancer Principles in Rule.Making
Origins of the Cancer Principles 

1. Extrapolation from Rodents to Humans
2. Can All Chemicals Cause Cancer?
119



121162

164

IQESZ

 

IV.

VI.

3, Do Benign Tumors Characterize a
Carcinogenic Response? .

4. Can "No-Effect" Levels be Set for Chemical
Carcinogens?

J. Organized Opposition
K. Attempt at Peer Review

CONCLUSIONS

A. Nature of Controversy -
B. How the Controversy Might Have Been Averted

NOTES

APPENDICES

120

 

165
168

172
176

184

184
186

188

204




BBSUUOZHOU

?8583

 2 (f
many scientists in OPP had extensive toxicological
experience in industry, USDA, or FDA with substances that
induce reversible effects below a certain "threshold" of
action. Thus, they believed "noeeffect" levels should be
set for carcinogens as they are for substances posing acute
and certain other chronic hazards. Furthermore, many in OPP
believed rodents (mice eSpecially) were inappropriate animal
models for cancer testing.

Because cancer is such a poorly understood disease,
there are no unequivocal answers to a great many questions.
The principles were an attempt to summarize reasonable
presumptions concerning the design and interpretation of
studies to detect carcinogens. They sought to present in
simple terms the thinking of expert committees that had
addressed the problems of carcinogenicity testing for over
25 years. Not surprisingly, many scientists in OPP opposed
the use of the principles, and COO took the lead in
preparing the scientific and technical basis for the
Agency's position on carcinogenicity in cancellation
actions. Under other circumstances, OPP would have had a
large hand in the cancer issue during cancellation actions;
OPP and other Agency scientists testified on many issues
other than cancer.

U?snvoznou

I. Origins of the Cancer Principlele9

Although it is not the intention in this case study to
evaluate the scientific validity of the cancer principles,
it seems appropriate to examine in detail some of the
underlying documentation. For this reason, some of the
reports and articles that were entered into the record of
various fact-finding, cancellation, and suspension
proceedings, and that form the basis for the principles will
be discussed. The following discussion is irtended to
illustrate the surprising consensus--short of unanimity--on
the concepts embodied in the cancer principles and is
limited to four such concepts:

-- It is necessary to extrapolate from animal studies,
particularly in rodent species, to humans in order to
evaluate the risk of cancer.

Not all chemicals are capable of causing cancer, even
at maximum doses of administration.

Although a distinCtion can frequently be made between
benign and malignant tumors, for the purposes of
establishing the carcinogenicity of a substance, they should
be considered synonymous.

161

isas?

 



. -



There is nc practical method for determining a "no-

effect" or threshold exposure level in humans for
carcinogens.

1..

already been discussed earlier to some extent.? It should be
recalled that the various expert committees agree that
reliance on animal studies is necessary because of the
practical and ethical limitations in reliance on
epidemiological studies to identify carcinogenic agents and
because of the evidence of the validity of studies in
experimental animals as the basis for establishing the
carcinogenicity of a substance.

Extrapolation from Rodents to Humans

The first issue of reliance on studies in rodents has

159? WEI-IOU

Discussions by expert committees of the optimal

conditions for carcinogenicity testing has included
consideration of appropriate mammalian species for testing.
After a thorough review of carcinogenicity testing, the Mrak
Commission reported in 1969:

The species most practical for testing are rats,
nice, and--as more recently shown??hamsters.
Strains and colonies should be selected to provide
adequate sensitivity to tumor induction, as
revealed by positive control tests with known
The use of nonrodent Species,
recommended in the earlier reports, has now been
substantially dropped. A suitable, practical
nonrodent Species would be useful but it is not
available at this time..-. While dogs have been
employed for tests of carcinogenicity, with
noteworthy success in selected cases {bladder
carCinoqenicity of aromatic amines), the
requirement of lifetime feeding makes this Species
too expensive, in terms of time and funds, to be
employed routinely.11?

Again in 1975, an NRC committee confirmed the views

expressed in many of the earlier reports:

Rodents are the animal of choice for carcinogenesis
tests because of their convenience, comparatively
short life span and proven susceptibility to a
broad range of carcinogenic agents.?1

Another NRC report published in 1975 concludedi

carcinogenic risks, and other health
hazards, of pesticides require continuing 
evaluation by testing with laboratory mammals;
the problems involved in translating

162


.-. u. . -



. - . 
inif!? .

s!



Fr

 

 

 

 

 

-- .

 


103 lbid.

10? Federal Register, Vol. Q0, (July 3, 1975), pp. 
28296.

 

105 Ibid., p. 28256.

 

. :3

106 Ibid-, p. 28257. CD
107 Ibid., p. 28263. co
c:

108 Bob Wyrick, Officials Devised Cancer Tests on 4*
People," Washinthn Post, Thursday, June 23, 1977, p.4>

PrOposed Carcinogen TeSt on Mexicans," Ibid.E:
Wednesday, May 11, 1977, p. A1. 



10 I wish to acknowledge the contribution made to the
writing of this section by John E. Bonine, Office of
General Counsel, EPA. Bonine gave me a preliminary
draft of a paper tracing the term "principles" and
reviewing some of the reports of expert committees.

110 Mrak Commission (1969), supra note 37, p. ?65, text and
footnote 2.

111 National Research Council (1975) Evaluating Chemicals,
supra note 37, p. 150.

112 National Research Council (1975) Pest Control, supra
note an, p. 66.

113 Mrak Commission (1969), supra note 37, p. 11.

11M National Cancer Advisory Board (1977), supra note 31, p.
863. 

11S Tomatis, supp; note 37, p. 1.

116 0.8. Department of Health, Education and Welfare, ?urvey
of Compounds Which Have been Tested for Carcinggenic
Activity, Volumes (197d-1976), (Washington, D.C.:
Government Printing Office).

117 The study was performed by the Litton-Bionetics Research
Laboratories under contracts PH u3-6u?57 and PH u3?67-
735; see 0.8. Department of Health, EducatiOn, and
Welfare (1969), supra note 31, pp. ?61*506; J.R.M.
Innes, B.M. Ulland, M.G. Valerio, L. Petrucelli, L.
Fishbein, E.R. Hart, A-J. Pallotta, R.R- Bates, H.L.
Falk, J.J. Gart, M. Klein, I. Mitchell, and J. Peters,
"Bioassay of Pesticides and Industrial Chemicals for
Tumorigenicity in Mice: A Preliminary Note," J. National
Cancer Institute, Vol. Q2 (1969), pp. 1101-111U.

 

198

.16696

(W 2626

SUMMARY OF RESPONSE
TO RISK ANALYSIS ON

THE USE OF HERBICIDE

compiled by

surgeon?

Marguerite L. Leng

Product Registrations
Health 6 Environmental Sciences
THE DOW CHEMICAL COMPANY
Midland, Michigan USA

February 1979

Excerpts from Volume I of Dow risk rebuttal
and from Dow legal brief
submitted to U.S. Environmental Protection Agency

on August 4, 1978

-??z?212 H00

18537

$5231

 

 

 M. it Wm?, 23 zen/Ira 
. U. ALu?-dwei 5 ?104   am; 
7 ?lm 7/5/07 Wn?aq?r
APPENDIX II 7% 
- 'Ma, 
ggxuzsf 
o~014 .anmaqn?aum?lan 

 

Lutturu Submitted to EPA In ?ghponuu LO Nu Ill Effocte from Huqlgturcd Unuu.

 

EPA I Name of . Egan gaggpatlon I 'Yunrs of use
5 Jumus Hruln Husehnrg 0n Sucrutary, Stutu Sprayura 400 man yours
6 Mack L. Little Lufkln TX runchur/aprayur slncu 1958
7 C. Eddie Muuku Thumnatun an County ugunt coup uxtunulon

1] Donald H. Cuutullnu Huncuw ID pcut control consultant 20 years
10 Huwurd G. chouull I Hlsnoulu MT Exec. V.P. Inland Forest

Resource 

20 J. r. Howell Clnrundun TE rancher yearn

34 John N. Grunt County MN County extension agent yuaru

J7 Frcdurlck Drummuud Puwhunku 0K uprayer/ranohcr yearn

4] Hm. J. Lubbock TX t?uucarcll and tango manngor JO yuuru

17M A. Hu?ougul I National Cuttlomnn'? huaoc. . (280,000 profonslonnla}

D6 James L. Colo Leavenworth In small ranch and uuodu nvur 25 yunru
llnn?y Hahn: (2A numagur, uoudlmulu rnnuarch yearn

9l Jumun H. Floyd lenrnl KS nurlul 200.000 acres

9] Patrick J. Runny Huutborough MA New England Power Survlcu Co. 20 yearn
102 Hunnld A. chhlell D.C. Exec. Director Publlc Lands
Council

. KOO

IJOHMJ 


I

 

 

TRANSLATION
- --.I C:
GERMAN Publication 9/979 
nut-Il?
9 on Period April 29, 1981


z: 

0 Lu
mm
OD


of.the Federal Government c:

to U1

the inquiry of members of the parliament, Frau Dr. Hartenstein,
Schafer (Offenburg), Catenhusen, Duve, Jansen, Kiehm, Dr. Hub er,

1
Marschall, Frau Dr. Hartiny-Glotz, Reuter, Dr- Schwenk (Stade)
Frau Weyel, Hitek, Frau Zutt, Ibrugger, Bargerowski, Engel?
hard, Dr.Hirsch, Kleinert, Dr. Hendig, Wolfgramm (Gettingen) and
the parliamentary groves 3f SPD and EDP

Publication 9/292 gh/yl - 

/4x4 a%%2?mf j?amazlasu a254;?(

 



 



- I 



The Secretary of the Dept. of Food, Agriculture and Forestry
has.answered the inquiry on behalf of the Federal
Government by letter of April 14, 1981, as follows:

The Biologische Punoesanstalt Jar sand? und (BEA)
and the BuneesgesunuheitSam: CA) V?lch are responsible for the
registration of pesticides, have repeatedly xpressed their expert
Opinion on the herbicide 2,4,5?1. It was established in a joint
statement of the year 1975 that a hazard does not exist if it is
used properly and according to the regulations. Necessary restri?_
tions for the use are ruled by the decree about prohibitions and
restrictions of the use of pesticides, dated December 19, 1980
3031. p. 2355) or by requirements

2,b,5?t containing herbicides as registered in the Federal Republia
of Germany today, cannot be put in a relation to the "agent orange:
which was used as a defoliant for forests in the Vietnam war. Thesr
products were contaminated very much with the toxic TCDD (Dioxin),

which also reached the environment during the accident.ofjseveso,
The defoliant used in Vietnam contained up to 5000?times more TCDD
than the formulations registered in the Federal Republic of German;
today. The EBA has required from the manufacturers in the registrai
that a TCDD content of 0.01 mg/kg is not exceeded, i.e.

100 g/t In contrast to this, the guideline level of the
world Health Organization amounts to 0.5 mg/kg, i.e. SO?times highc
than the German guideline level. 1635 ?g
2 

 2832:}

  .- 



serge-er a re 

Jr.? 
.I-I

A symposium sponsored primarily by the American Chemical Society's 
Division of Pesticide Chemistry, with the cooperation of the Committee 
on Chemistry and Public Affairs, and the Divisions .of Agricultural and- a 
Food Chemistry, Chemical Health and Safety, and. Professional   .
Relations. Held on 1980, in Las Vegas, Nevada, during the 
180th national meeting of the Society.

 American Chemical Society

1155 16th St., NW.
Washington, DC. 20036

 

 



2644

AIEEHHIEX 
C.

PROPOSALS FOR EPIDEHIOLOGICAL INVESTIGATIONS OF 
HORBIDITY ASSOCIATED HITH HERBICIDES 

The assessment of the available data concerning a possible biological 
association between exposure to phenoxy acid herbicides and soft tissue .2
sarcoma has led to the conclusion that there is a need for further research 4:

to permit the confirmation or refutation of this suggestion. It has also
been noted that a number of studies are already in progress or are about

to be initiated in several countries around the world using data which

are available on both a national and international basis. These include
systematic epidemiological studies of both a retrospective and a prospective
nature which constitute the only appropriate methods available for this
purpose. In view of the known temporal and geographical yarintions in the
incidence of a number of malignant conditions and of the importance of the
phenoxy acid herbicides to agriculture and forestry in this country it is
considered advisable to investigate the situation as it exists in the

United Kingdom. Appropriate information is available to enable epidemio-
logical studies of both a retrospective and a prospective nature to be
carried out. In addition to being the only way of obtaining information

of direct relevance to the UK workforce it should be possible, from the
data available, to obtain evidence which is less open to the criticisms

that have been levelled against some of the reports that have been published
so far. There would also be an opportunity to extend the scope of the
investigations to include aspects other than the occurrence of soft

tissue sarcoma and the possible involvement of other chemicals such as

the chlorophenols.

Retrospective Study

This would examine whether patients suffering from soft tissue sarcomas (or
other specified malignant conditions) had an unusual history of exposure to
phenoxy acids during their manufacture or use as herbicides. The first

step would be to establish a group of sarcoma patients and an appropriate
control group. Since comprehensive cancer registration data are available
covering the whole country, basic information eg age, sex, type of
malignancy, when and where registered, for all identified cases in

specified categories of soft tissue sarcoma is readily accessible. However,
these are relatively rare tumours, and a decision would have to be taken
whether to study all of them in relevant age groups, or only those in

parts of the country where herbicide use?is likely to he more common.

The control group, which would have to be matched at least by age and

Sex, could be cancer registrations for body sites where there are no

reasons to believe that tumour occurrence could be associated with herbicides.

The major problem with retrospective studies is in obtaining the history of
herbicide exposure. For most patients the occupational exposure will

out have been documented precisely, if at all. Hemories are notoriously
fallible, even in the short term and recall concerning events many years

16601.

 

previously will probably be uncertain. For patients who have died recourse
might be had to their families to obtain the history, as in some of the
published reports, but this would be even.more unreliable. Furthermore,
whether obtained from the patient or a relative, there is the possibility
of a bias in the histories of herbicide use from cases compared with
controls because of the publicity on possible hazards.

A retrospective study would have to be conducted in the eXpectation that
only in a small proportion of cases and controls could the histories of
herbicide use be validated. In such an investigation particular attention
would have to be paid to ethical considerations. Information in cancer
registries is confidential, and a proper procedure would have to be?
established before any approach could be made to consultants, general
practitioners, to the patients and their families, or to any other source

of data.

Despite these drawbacks a retrospective study has certain advantages. It'
I can be mounted fairly quickly and is relatively cheap. It does not take
long, the duration being governed largely by the time required to obtain
occupational histories. 

QHZZIZ ma

Prospective Study

In this study a cohort with known exposure to herbicides would be identified
and monitored to determine whether they suhsequently.3uffered any excess

of soft tissue sarcomas or morbidity of other kinds. There would be a

need for a control cohort comprised of individuals not exposed to herbicides.
The subsequent morbidity of this group would be recorded in exactly the

same way as for the herbicide user cohort.

Since reliable information concerning exposure is of paraaOunt importance in
such a study an essential pre-requisite is that there should be good
documentation available concerning herbicide use. Organisations such as

the Forestry Commission or commercial sprayers who keep rOutine records
should therefore be selected in preference to farmers whose occasional use
of herbicides may go unrecorded. The documentation system would have to

be reviewed to assess the need for possible changes to ensure that

complete, uniform and appropriate data are obtained.

The essence of the prospective study would be the collection of morbidity
information in cohorts defined as either using or not using herbicides.
Leaving aside the choice of the control cohort, the non-users, there are
different problems depending on what aspect of morbidity is selected for
study. The extreme event, death, could be readily ascertained, even if
members of the cohort moved or changed their occupation, by tagging their
entries in the was Central Register. Cancer registrations, as well as
deaths from any cause, are reported to this Register so that the cancer
morbidity as well as mortality can be obtained for the defined cohort.

Other expressions of morbidity, leading-to consultation with a general
practitioner, out-patient referral or admission to hospital, are nut
obtainable from a central register. If there was a case for seeking
evidence on anything besides general mortality and cancer morbidity,

Lassa

10

11

12



ag -on liver disease, this would need a separate organisation such as

a questionnaire to the patient or his doctor to determine whether there
was any episode of illness worth further enquiry. It would add considerably
to the cost and complexity of the study to extend the_range of outcomes
in this way, and there would need to be a strong case for such addition.

HUG

It could be difficult in a prospective study to establish a satisfactory
control cohort. Ideally it should comprise individuals of similar age,
sex, and social class distribution, and probably from the same areas. In
occupational studies of this kind it is sometimes possible to find groups
within the same organisations, such as administrative and clerical staff,
who are not in contact with the process, but their numbers-may well he
too small in this instance. It is always possible to use national or
Regional data on mortality and cancer registration as a standard of
comparison for the cohort of users, and this compromise may be a reasonable
one to adopt. 

SHZZIZ

The main disadvantage of a prospective study is the length of time before 
sufficient events have occurred to warrant comparisons betw users and
non-users particularly where rare events such as the occurrence of soft

tissue sarcomas is concerned. Furthermore, the cohorts defined initially

will not remain 'pure' in the sense that additional exposures to similar

or different agents may occur. Nevertheless this type of study is more

likely to lead eventually to a valid measure of the risk, if any, of

exposure to specific herbicides and should, therefore, be given serious
consideration, although the difficulties which have been Outlined above should
not be underestimated.

A cohort, prospective, study set up now could provide evidence only on
phenoxy acid herbicides currently produced. However, there may be scope

for conducting a cohort study retrospectively. If in selected organisations,
such as the Forestry Commission, it was possible to prepare nominal lists of
all employees in some period in the past, say the decades {945-54, 1955-64,
and to categorise them by pesticide use, their subsequent experience up

to the-present day of mortality and cancer registration might be retrievable.
The aortality picture could be virtually complete, but cancer registration
was not fully established in the earlier years and would be reliable

only for the last 10-15 years. The study would not be easy, but might be
valuable because it would provide information on exposure to herbicides

with a relatively high'dioxin contamination. Unlike a new cohort study
established now, some measure of the outcome 20-30 years after exposure
could be obtained relatively quickly. 

166-03

SUMMARY OF EPIDEMIOLOGICAL AND CLINICAL STUDIES CONTRIBUTING ORIGINAL
OBSERVATIONS CONCERNING A POSSIBLE ASSOCIATION BETWEEN CHEMICAL

2645

EXPOSURE AND SOFT TISSUE SARCOMAS


if: . .Origin

CASE CONTROL STUDIES

Study Population



 

52 cases of soft tissue sarcoma
and 208 latched controls from
Northern Sweden

1 'Suedsn

 

A.
C:
CD
he
Summary of findings ?d

LO
CD

Exposure to phenoxy acids reported
by 252 of cases and 72 of controls.
Exposure to chlorophencls reported
by 132 of cases and 32 of controls.
Relative risk* of developing a soft
tissue sarcoma associated with
exposure to phenoxy acids - 5.3.
(Relative risk associated with 
exposure to chlo 'whenols - 6.6).
Chlorinated dibenzodioxins and
dibenzofurans possibly involved.

*Relative risk is the statistically derived expression of the ratio between the inciden
of exposure reported by cancer patients and that reported by the control group.

110 cases of soft tissue sarcoma
and 219 matched controls from
Southern Sweden.

2 Sweden

3 Sweden 169 cases of malignant 
(60 Hodgkin's disease and
109 non-Rodgkin's and
333 latched controls.

4 Sweden 15? cases of colon cancer and
5&1 unmatched controls.
(Supplementary study to
l, 2 and 3 above).

Exposure to phenoxy acids reported
by 131 of cases and 3.23 of controls
Exposure to chlorophenols reported
by lOZ.oi cases and 3.7: of controls
Relative risk of developing a soft
tissue sarcoma associated with
exposure to phenoxy acids - 6.8.

.(Relative risk associated with
"exposure to chlorophenols - 3.3).

Exposure to phenoxy acids reported
by 242 of cases and 73 of controls.
Exposure to chlorophenols reported
by 301 of cases and 10: of controls.
Relative risk of developing a
associated with exposure

to phonoxy acids - 

(Relative risk associated with
exposure to chlorOphsnols - 4.3).

A re-examination of the data
presented in l, 2 and 3 above
confirms relative risk factors of
4.2, 5.6 and h.l respectively for
an association between the develop-
ment of soft tissue sarcoma and
malignant and exposure to
phenoxy acids follOUlng a 5 year
latency period. No association was
established between expGSure to Such
chemicals and the development of
colon cancer, relative risk - 1.3.



Ref

Countrv of

Origin
5 New Zealand
6

7

8

New Zealand

COHORT STUDIES

West Germany

Finland

USA

Study Papulation

 

102 cases of soft tissue sarcoma_

first diagnosed beLvecn 1976
and 1980 and 306 other cancer
patients as matched controls.

80 cases of soft tissue

sarcoma and 92 other cancer
patients.as controls. (Further
data fro? the study reported

at 5 above).

75 chemical workers involved
in a trichlorophenol plant
accident in [953

L971 herbicide sprayers
(Railway, forestry, highway
and electric company

workers)

61?chemica1 workers involved
in trichlorophenol production
and exposed to 
in 1964 -

'6 due to cancer (4 expected).

PEPE-.1 A.

Summary of findings

 

Preliminary report. 0n the basis
of Cancer Registry Records there

is no evidence of an excess of soft
tissue sarcomas among the
occupational group involving
agriculture and forestry.

Further preliminary report.

Exposure to phenoxy acids reported
by 262 of soft tissue sarcOma

cases and 212 of controls. Estimat.
for the relative risk of

developing a soft tissue sarcoma
following exposure to phenoxy acids
varied from 1.3 for potential
exposure of more than 1 day at

least 5 years before cancer regis-
tration to 1.6 for probable expostg;
No soft tissue sarcoma patient had:;
worked full time on phenoxy acid 
herbicide spraying.

42 cases of severe chloracne
associated with exposure during
the accident or subsequent cleaanE
uork.? By 1978 17 deaths had
occurred (11 to 25 expected),

EHZIZ

The
incidence of stemach cancer (3 cases
was higher than expected. There
were no cases of soft tissue sarcoma

Retrospective study 1955-1971, 49 de
occurred (87.8 expected), 13 due to
cancer (16.4 expected). Prospective
follow-up, 1972-1976, 62 deaths occu
(92.9 expected) 9 due to center (18.
expected). Prospective follow-up,
1977-1980, 82 deaths occurred, (91.1
expected), 17 due to cancer, 18.4
expected. No cases of chlurucne. Hg
cases of soft tissue sarcoma.

49 cases of chlorecne were -rted
ranging from mild to severe. by
1978 4 deaths had octurred (7.8
expected), 1 due to cancer (1.6
expected). One case was a soft
tissue sarcoma (fibrusarCUma).


bL'l

 

11

12

13

14

15

16

222513.
Origin

USA

USA

Sweden

Sweden

Czecho-
slovakia

USA

USA

Study Population

20? chemical workers involved
in production between
1950 and 1971

121 chemical workers involved
13'! trichlorophenol plant
accident in 1949

142 forestry workers exposed
to phenoxyscetic acid
herbicides between 1954 and
1978 and 244 forestry
workers not so exposed

348 railway'workers (herbicide
sprayers between 1957 and 1972)

55 chemical workers exposed

to over a 10 year
period during the production

of 2.4.5-T from trichlorophenol

Review of 9. 10 and 11 above
plus additional data on
chemical workers involved in
the production of from
trichlorophenol

Supplementary data concerning
the cohort studied at 9 above

APPEIHIEX A

_Sumnary of findings

 

No case of chloracne. By 1976
11 deaths had occurred (20.3 expected)

-only l-due to cancer (3.6 expected).

This was not a case of soft tissue
sarcoma.

A11 menbers of the selected cohort
exhibited chloracne attributable 
exposure during the accident. By 
1978 32 deaths had occurred (46.6

expected), 9 due to cancer

(9 expected). One case was a soft
tissue sarcoma (malignant fibrous

histiocytome).



The pattern of-deaths among both
exposed and unexposed cohorts is in
accord with Swedish national
aortelity statistics. However
among foresen, the cost extensively
exposed workers, there is an excess
of eortality (8 observed/4.1 expected)
due to an excess of both cancer

(3 observed/1.0 expected) and
cardiovascular disease (4 observed/
1.9 exported). There were no soft
tissue sarcomas.

No cases of chloracne. By 1978

45 deaths had occorred (49 expected),
17 due to cancer (11.9 expected).

No specific type of tumour
predominates and no soft tissue
sarcoma was reported.

52 cases of chloracne. By 198?
6 deaths had occurred, 2 due to
cancer.. There were no cases of
soft tissue sarcoma.

An additional case of soft tissue

.sarcoma (generalised liposarcoma)

is reported in a worker involved in
the production of from
trichlorophenol. There was no prior
history of chloracne.

An additional case of soft tissue
sarcoma (malignant fibrous
histiocytoma) in a trichlorOphenol
production worker is reported.
There was a previous history of
chloracne.

.1668?

18

19

20

21

22

23

Oodntr of
Origin

United
Kingdom

USA

USA

Study Population

 

126 chemical workers involved
in a trichlorophenol plant
accident in 1968

Retrospective analysis of
mortality data - Hashington
State (1950?1979)

Retrospective analysis of 
mortality data - butchers end
slaughterhouse workers
(1965-1980)

CLINICAL REPORTS

Sweden

Sweden

USA

USA

7 cases of soft tissue sarcoma
(included in the case control
study reported at 1 above)

17 cases of malignant 
(included in the case control
study reported at 3 above)

1 case of soft tissue sarcoma
in a chemical worker_

2 cases of soft tissue sarcoma
in chemical workers (father
and son)'

.APTEIIDIXI.X

Summary of findings

 

79 cases of chloracne were reported

after the accident. By 1981 no
deaths had occurred nor had any
case of cancer been identified.

No abnormal biochemical or clinical
effects attributable to exposure.

In occupations with exPosure to
phenoxy acid herbicides or
chlorophenols ?9 soft tissue

sarcomas were observed (41 expected).

In those occupations with the

greatest excess of deaths from

soft tissue sarcoma there was 
obvious association with exposu
to phenoxy acid herbicides or
chlorophenols.

An analysis of cancer mortality
aoong butchers and slaughterhou
workers revessled an increased
risk relative for soft tissue

0
re

56

sarcoma of 5.6 (1 observed case/

0.13 expected).

Exposure to
reported in
5 cases the
and heavy.

phenoxy acids is
all 7 cases. In
exposure was direct

phennxy acids, is
8 cases.

Exposure to
reported in

A case of retroperitoneai
neurogenic sarcoma is reported

in a truck driver and maintenance

worker in a chemical plant
producing trichlorophenoi or
Ho history of prior
chloracne.

Two cases of soft tissue sarcoma
are reported (a flbro-sarcomatous
mesothelioma and a liposarcoma).

Both individuals had been
employed at a chemical plant
producing chlorOphenois. No
history of prior chloracne.


as?






IHZZIZ

l.

3.

l.

5.

6.

9.

10.

11.

REFERENCES

Hordell, L., (1979). British Journal of Cancer 32 711-717
"Case-control study: soft-tiesue sarcomas and exposure to phenoxy
acetic acids or chlorophenola".

Eriksson, H., et a1.. (1931). British Journal of Industrial Medicine
2??27-33. "Soft-tissue sarcomas and exposure to chemical substances:
a case-referent study".

Hardell, L., et al., (1981), British Journal of Cancer 32 169-176,
"Malignant and exposure to chemicals, especially organic
solvents, chlorophenols and phenoxy acids: a case-control study".

HUD

Hardell. L., (1981) Scandinavian Journal-of Work ?nvironment and
Health 1 119-130 "Relation of soft-tissue sarcoma, malignant 
and colon cancer to phenoxy acids. chlorOphcnols and other agents?.

Smith, ML. et al.. (1932) Community Health Studies 3 114-119;
?Do agricultural chemical cause soft tissue sarcoma? Initial findings
.of a case-control study in Neu Zealand". 

Smith. A.H., et a1., (1982) Proceedings of the 3rd International
Symposium on Chlorinated Dioxins and Related Compounds. In Press.
"The New Zealand soft tissue sarcoma case-control study: interview
findings concerning phenoxyacetic acid exposure".

Thiess, A.H., Frentzel-Beyne. R., (1978) working paper, quoted in
Internation Agency for Research on "Long-term hazards
of dihenzodioxins and dibnnzofurans?
Internal Technical Report No 78/001. Lyon. "Hortality study of persons
exposed to dioxin after an accident which occurred in the BASF on

13th November 1953".

Riihimaki. V., et al., (1982) Scandinavian Journal of Hork Environment
and Health 37-h2 "Mortality of 2,h-trichlorophenoxyacetic acid
herbicide applicators in Finland".

Cook, R., et al., (1980), Journal of Occupational Hedicine 33 530?532
"Hortality experience of employees exposed to 
dihenzo-p-dioxin 

Ott, HG., et al., (1980) Journal of Occupational Medicine 5; 

mortality analysis of employees engaged in the manufacture of

2,4,S-trichlorphenoxyacetic acid".
216k. Suskind. R.R., (1980) Jaurnnl of Occupational Medicine

3; ll-lh ?The mortality experience of workers exposed to Lutrachloro?
dibenzodioxin in a trichlorophenol process accident".

18608

12.

13.

16.

15.

16.-

17.

18.

19.

20.

21.

22.

23.

Hogstedt, 6.. Hesterland. 3.. (1980) L?kartidningen 12 1829-1831
?Cohort study of causes of death of forestry workers with and without
exposure to phenoxy acid preparations".

Axelson, 0., et 31., (1980) Scandinavian Journal of Bork Environment
and Health ?_73-79 "Herbicide exposure and tumour mortality. An
up-dated epidemiological investigation on Swedish railroad uorkers".

Pazderova?Vejlapkova. J., et al., (1980) Pracovni Lekanstri 204-209
?Chronic poisoning by 

Honchar, 9.1., Hnlperin, 3.8., (1931) Lancet 1 268-269 
trichlorophenol, and soft-tissue sarcoma".

Cook, R.R., (1981) Lancet 1 618-619 "Dioxin, chloracne, and aoft-tissuq:3
sarcoma". - '1

. 

May. 6., (1982) British Journal of Industrial Medicine 22 128-135
?Tetrachlorodibenzodioxin: a survey of subjects ten years after no
exposure". - 
. ho
Hilhau. 5.. (1982) Lancet i 1464-1545 "Herbicides; occupation. and 
cancer". 
4r
(.0

Johnson, 3.8.. Fischean. H.R., (1982) Lancet 1 913 "Cancer mortality
among butchers and slaughterhouse workers".

Hardell. L., (1977) L?kartidningen "Malignant 
tumours and exposure to phenoxy acids - a clinical observation".

Hardell. L., (1979) Lancet 1 55-56 "Malignant of histiocytic
type and exposure to phenoxyacetic acids or chlorphenols.

Hoses, H., Selikoff, I.J.. (1981) Lancet 1 1370 "Soft tissue sarcomas,
phenoxy herbicides and chlorinated phenols".

Johnson, al., (1981) Lancet ii 3&0 "Soft tissue sarcomas
and chlorinated phenols".

l-J

man-L?E' -

 13th Decenber, 1982



Minister of Agriculture, Fisheries and Food,
Whitehall Place, - 
London SW1A ZHH.

The Rt  Peter Walker, LEE, MP,  

Hm

Dear mister 

1. In your letter of 3 May 1982, you sought the Advisory Ccmnittee on
Pesticides' observations on new evidence concerning the safety of 
herbicides, with particular reference to a review article published

in the Lancet on that ?ne date. We have now reached a point in our
enquiries at which we think it would be helpful to report to you and

the other Departmental Ministers with an interest in the safety of

these herbicides. -

:400

Background to rStudy

2. The Committee decided that a panel of independent members should
be set up under my to consider the latest evidence,

and the advice in this letter reflects their detailed examination of
all the relevant studies known to us which are sunmarised in Appendix
A to this letter. The panel included all areas of expertise needed
for a study of a postulated association between use of a herbicide
and a particular typeof cancer, and consisted of:- 

11912212

Professor C.L. Berry
Dr R.L. Carter

Dr R. Goulding
Professor .3. Knowelden
Professor R.I. McCallun
Professor G.R. Sager

3. Before giving you our conclusions, I think that a. brief review of the
evidence we have considered since publication of our Deoen'uer 1980

report might. help you and your colleagues to assess our advice. You

will recall that much of our 1980 enquiry was concerned with the question
of whether the use of herbicides might be associated with
miscarriages and birth deformities among humans and animals, and the evidence
included a umber of specific cases in which such an association was
claimed. Each of these was investigated in detail by the appropriate
Governnent Departments and the Health and Safety Ebcecutive in consultation
with the general practitioners, hospital consultants and veterinary
sn'geons concerned, before the Cozmittee reached its conclusion. Our
examination revealed no evidence linking any of the cases with use of
herbicides: and we could find no other evidence which established
any association between use of these products and harmful effects on
humus, other living creatures or the environment. We were thus able

to conclude that: herbicides can safely be used in the UK in
the reconmended way and for the recomended pun-poses".

4. Our present enquiry arose out of a separate and specific question -
that there might be an association between exposure to phenoxy acid
herbicides, chlorophenols or the contaminants that may be present in
these products and substances, and an increased incidence of a group

of manure classified as soft time soreness. The term soft tissue
sarcanas includes some 50 varieties of tlmour. As a group they are

rare, in that they represent approximtely one in every 1500 indiViduals
who die each year, and three from every 1,000 people who die from

cancer each year. Obviously such a low overall incidence makes assessment
of changes related to possible causal agents a most difficult exercise in
statistical significance.

IT

@8510-

5. The evidence we have reviewed is based on studies of two quite
distinct types of populations. The first is of users of various herbicides
and wood preservative products, and the second is of factory workers
engaged in the production of chemicals. The ACP is concerned with
advising on all risks that may arise the use of pesticides, as
distinct fran any risks which may arise in the course of manufacture, and
which are strictly speaking outside our remit. We, therefore, concentrated
on the first type of population - pesticide users a- and our advice is
based on the weight of evidence emerging for this type. Nevertheless,

we also examined factory studies to ensure that no evidence which might
have a. bearing on our task had been overlooked.

Discussion of Evidence - Herbicides and Wood Preservatives

6. Studies of pesticide user groups have been undertarzen in various
countries and consist both of case control and mortality investigations.

The case control studies involved a canparison of individuals with soft tissue
saroanas whose exposure to herbicides and wood preservatives had been
assessed mainly by memory recall, with control groups who either had no
cancer or other types of cancer. In the nortality studies the incidence

of death among groups of workers largely identified on the basis of
occupational histories was canpared with national or regional unrtality
statistics. -

HOG

SEIZZIZ

7. For users of herbicides and wood preservatives, a positive statistical
association between reported exposure and soft tissue sarcums had been
sugested from two studies carried out in Sweden. Successive reports on

a similar case control study have also becane available frun New Zealand.
These showed no correlation between exposure to phenoxy acid herbicides and
'soft tisme sarcomas. It is our view that the procedures used for
establishing the Swedish study groups, and some aspects of the collection
of the exposure data, were not reliable. In that country, separate

case control studies were undertaken in Northern and Southern SXveden
following publication of a previous clinical report in which 7 individuals
with soft tissue sarcanns recalled being exposed occupationally to

nhenoxy acid herbicides at some time during the 1950's and 1960's. I
should explain that whenever clinical evidence of that kind becomes
available in any area, it is normal epidemiological practice to follow

up the findings by means of investigations anong totally different

groups, thus preventing the possibility of bias. In the follow up

study in Northern Sweden, however, the 13 cases in which exposure to
phenoxy acid herbicides was claimed a total of 52 cases of

soft tissue sarcanas) included the 7 cases identified during the earlier
clinical investigations. 'u'e consider this to be unorthodox epidemiological
practice; if these 7 cases are emitted from the study, the relative

risk is reduced to within error limits. r[his was not the only defect

in the study. For example, patients and controls who reported exposure
only to chlorophenols were excluded from the calculation of the relative
risk factors associated with phenoxy acid herbicides while the control
group excluded patients with any malignancies. The relative risk factor
for chlorophenols was similarly artificially raised by excluding patients
and controls who reported exposure only to phenoxy acid herbicides. lbreover,
that study, and the further one in Southern Sweden, were conducted at
least one year after a publication drawing attention to the possibility

of the suggested association from clinical experience. BeCause public 
awareness of alleged risks from herbicides had been acute for

sane years, there is a probability that a bias from exposure recall
occurred in both studies. .

2

?18811'

8. Although. they may be valuable, case control-studies are not a

precise tool for establishing a causal relationship. They can be very

unreliable and misleading unless supported by sound demented

infomtion on exposure. Such information was not available in either
oftheSuedishstudies, mere itwaslargelybasedonthemmry recall

of individual patients or their relatives. We have, therefore, looked

for corroborative evidence from mortality studies of cohorts of herbicide

users. We have found none. The such cohorts have been studied in

Shades and one in Finland. A total of 2,461 individuals were included

in them. There was no overall increase in deaths Iron all causes or

from cancer. No case of soft tissue was identified. Similarly, t:
- no correlation betveen soft tissue sarocnas and ocmpations involving
the use of phenoxy acid herbicides energed a study of deaths in 
Washington State between 1950?1979. We content further on this study in
paragraph 13 below. 

9. Che feature of our 1980 enquiry again became evident during the
current one and has already been referred to above. This is that much
of the evidence depends on personal recollection of what took place well
before the association was claimed, and that recollection may have been
in?uenced by later publicity. In our 1980 report?ue argued, in relation
to miscarriages and birth deformities, that it can be relatively easy

to select sane products, or other factor, and without enquiring very
closely, to assure sane connection with family unsfortxmes of that kind.
'Ihis tendency has again been illustrated by a recent publication in this
wintry ("Portrait of a Poison: The Story") in which two

deaths were attributed to soft tissae samcmas developing after alleged
exposure to herbicides. We have enquired carefully into these
cases, which have been given some publicity. In addition to the lack of
reliable exposure data, we have established beyond doubt that in neither
case was the tumor involved a soft tissue sarcma?.



Conclusions Herbicides and Wood Preservatives -

10. In summary, a umber of factors have emerged from our analysis

of studies of user groups that are relevant to the assessment of the
safety of herbicides in this country. none of the
reports we have examined is concerned only with use of formulated

herbicides, or indeed, only with production of These
studies relate to the phenoxy acid of herbicides as a whole, as
well as to seed preservatives. In this country, use of herbicides
constitutes only a fraction of the total use of all these

products. Secondly, deficiencies in those studies that gave positive
results are such as artificially to raise the calculated relative risk
factor. And thirdly, we-are not aware of any instance in this country
where an individual has used herbicides and deve10ped a soft tissue
scream. We conclude that there continue to be no grounds for changing
our previous advice that foundations of herbicides as

presently cleared, or, for that matter, other phenoxy acid herbicides

and related wood preservatives, do not pose a safety hazard, whether

used in agriculture, forestry, the bare and garden, or elseuhere.

11. Hmever, it has always been our nor-ml practice to keep the safety
of any pesticide under continuous review, and to take account of any
additional relevant evidence that may becane available. In the present
case, we believe it would be practicable and desirable to pursue scne
further enquiries, based on available UK cancer mortality data, in order
to establish whether there has been any change in the incidence of

soft tissue sarcomas over the past 20 years, and whether there has been
any correlation betw?en such sarcanas and particular occupations.

This will take'a little unre time, but without such an analysis the
summation we havemde of all known relevant studies would not be
conplete. We will, therefore, be reporting further.

3 1651

12. Separately from this we suggest that the Covenant should consider
arranging for systematic and canplernentary retrospective and prospective
case control investigations to be undertaken as a contribution to the

work being done internationally in this area. To this end we set out,

in Appendix B, details of the kind of enquiries which would be appropriate
and amnent on the problem involved in setting them up, and on their
potential value. . .

Discussion of Evidence - Production of Chenicals

13. The second type of evidence we have reviewed (paragraph 5 above)
comes from mrtality and disease data of cohorts of chemical workers in
the USA, West (lemony, Czechoslovakia and the UK. Whereas in the last
three of these countries the findings were negative, positive evidence
has emerged only ?rm the USA, 1where 7 individual cases of soft tissue
sarcmzas have been identified in various studies. Three of these cases
were the subject of individual clinical reports where neither the size
of the exposed population nor the nature and extent of exposure, if any,
of the patients was established. Another case occurred among an
undefined group of chenical production workers. ?the remining three
cases, each of which had a prior history of chloracne testifying to the
fact and intensity of toxicologically significant exposure to dioxins,
were drain: from two cohorts carprising a total of 182 chemical production
workers. Exposure to chlorophenols was ccmnon to the latter 4 cases.
Other cohort studies in the USA and elsewhere among workers similarly
encposed to chlorophenols have revealed no cases of soft tissue sarconns
amng a total of 460 workers. Two further American studies, involving

the retrospective analysis of mortality data, did not establish any
association betwen the incidence of soft tissue sarconas and likely
occupational exposure to either phemxy acid herbicides or chlorophenols.

Of]

ZEIZZIZ 

14. We do not consider these studies to be relevant to any assessment
of the safety in use of the phenoxy acid group of herbicides, because the
nature and extent of possible exposure differ greatly
between the factory and the field, the forest or the garden. The studies
of cohorts of chemical prediction workers which we have examined,

and which identified the occurrence of chloracne among sons of the
individual cases, indicated that exposure was to both a wider range

of substances and to higher levels than was the case with users of
fonmlated products, and that it included ezqosure to contaminants

which have now been virtually eliminated fonmlated products.

There was no such evidence amng the populations who had merely used the
formuated products.

Conclusion - Production ..of manicals

15. In this country, chlorophenols are produced and used in many
situations outside agriculture and wood preservation. A few have limited
use in their own right, but the unjority are used, or have been used,

in the production of a wide range of products such as adhesives, dyes,
paints, inks, disinfectants, wax emlsions, cutting oils,  extinguishers,
ropes, tents, pharmaceuticals, toothpastes and cosmetics, as well as in
the leather, textile and paper and board industries. The evidence we
have examined concerning chemical production workers leads us to the
conclusion that it would be prudent for the Government to undertake
further relevant investigations in factory situations in- this country.
Neither the factory processing of chlorophenols nor the safety of the
predicts and practices described above is within the remit of the -
Advisory Cannittee on Pesticides; but we would .wish to be involved

in any such wider investigations in so far as these might relate to the
specific area 2; the. production of herbicides and mod preseryatives.

Yours sincerely,
Robert Kilpatrick
Chairman

Advisory Committee on Pesticides 4

18813.

pm

2849
DOW CHEMICAL USA.

 

ROUTE 1. BOX 1313

August 28,1980 DAVIS. CALIFORNIA same

915-753-5500
J. B. SWEELEY .

W. E.

MASONITE CORPORATION . cc 3. H. 3:35:5an
WESTERN WOODLANDS DIV. D. D. Hunt
P.O. BOX 97 M. G. Norr'
CALPELLA CA 95-118 A. J. Watsc:
Dear Jack, 

Regarding your letter of August 12, I just returned from vacation hence
the belated reply.

Based on our knowledge of and the claim or insinuation that
these materials could cause any adverse effect on humans or animals at

the low exposure levels resulting from applications several years previously
is spurious. Hundreds of water samples have shown that the levels im-
mediately below the treated area are less than 10-15 ppb. Data from Norris
and others indicates that the levels are reduced to non-detectable ones in

)5 mile or so There is a tolerance for in potable water
for 100 ppb.

Man 

At the rate was applied, there would have been very little in the
water immediately after application and only if run-off .0ccurred in the
spring after treatment. If there was some TCDD in the it would
have been decomposed mostly by sunlight and if it did get to the soil or

duff it is so insoluble that it would be extremely unlikely to get to the water

by run-off.

it is interesting that workers in the Dow plant have not reported
chloracne during the 25 years of operation. Chloracne is the earliest ex-
pression of reaction to TCDD. Nor have any adverse effects been reported
by workers in the plants?. These groups have been observed very care-

fully.

You may want some references to support the above statements; a copy
of the CAST report on Phenoxy Herbicides and several others concerned
with phenoxy herbicides that may be of interest are enclosed.

Since it is very unlikely that the "itchy back" resulted from exposure to

the forest herbicides, we can look for other causes. If Mr. Johnson "swims"
in the creek that is too shallow to swim in, he may be exposed to poison
oak or a number of other plants that can cause skin problems. Of course,
many other things, like soap, cleaners, oils, etc., are known irritants.

ft
m:

AN OPERATING UNIT OF THE DOW CHEMICAL COMPANY

18814

J. B. Sweeley -2- August 28, 1930
It seems odd that the "itchy back" has only now become serious enough to
cause a complaint 7 years after the last application. You may recall

the Seveso incident where 24-4 lbs. of TCDD was distributed over 200 to
400 acres in Italy. Children and adults were not moved out for 2 weeks.
Many cases of serious chioracne were treated, but after 12?18 months all
cases had healed up; there have been no other observed adverse effects
above those in the surrounding populations. The may last for 2-3
months and the about 1: to 6 months. It's quite impossible for these
materials to last more than a year or 2 in the trees and it disappears in 2-
ll- weeks from soil. The spring treatments would have allowed these or faster
rates of decomposition because of warming conditions and good soil mois-
ture. The previous treatments should have enriched the microorganisms
and enhanced breakdown.

I will pass this on to other members of our staff who may have added comments
of value. I hope this analysis and the reports will help refute the allusion
of Mr. Johnson.

Yours very truly,



. E. Warren
Agricultural Products Department
Technical Service and Development

Enc.

cc: 3. Jones, DOW CHEMICAL U.S.A., Sacramento.
Matt Anderson, CFPA, Sacramento
Norman Parker, Western Helicopters, Newberg, OR.

kim

if??



18815

. Dowzosonnl 2550

I Dow Chemical Cd;

EPA Hearing
Submissions He: Cancellation - Volume 2

16. "Petition for the Establishment of Tolerances for the Pesticide
Chemical, on Raw Agricultural Commodities?, Sec. C:
Full Reports of Investigations re: 12/8/67:

(1) Summary of 11 tests on safety of below)
(2) Conclusion:

I ?more data on than is available, but indi-
cations are that two materials are comparable; acute oral
toxicity of both in lab animals is in same range, 300?

1000 mg/kg. 

a -long history of usefulness not produced evidence of

unusual toxicity.

A. "Acute Oral Toxicity of to Rats, Mice, Gu'?
I I Chicks", Rowe, McCollister and Spencer, (Dow, 19

(1) Single oral doses (administered by stomach tube) of 

S-T as a 10% suspension in olive oil, to mixed sex group

I of rats, mice and guinea pigs, 3% solution in olive oil
given to chicks. 

(2) of poisoning ataxia, myotonia; death at highest
dosage levels; doses ranged from 0.1 1.0 grams/kg.

(3) LD 50's: rats 0.500 grams/kg.

mice 0.389 

guinea pigs 0.381 

i chicks 0.310 . 

(U) significantly more toxic to chicks than to rats.

B. "Toxicity of 2.H-D and a report on their acute
and chronic toxicity in dogs", Drill and Hiratzka, 1953.
("There have been no the toxic prepertie:
of 

(1) Experimental Procedures: adult mongrel dogs of both sexes

 

1 18816

used, fed "Friskies'
used,

DOHZUBUQ

commcuwxial 2

?single oral dose administered in capsules.
autopsied after in days (acute studies).

Results of Acute Studies2,u_D:

12

I .

,n?D and 2,n,5_T

Survivors

?oral administration in capsules imbedded in gram piece
of commercial canned dog food, 5
(chronic studies).

days/wk for 13 weeks

mg/kg caused death in 2/2 dogs in 2?3 days fol?
lowing administration.

-250 mg/kg
?100 mg/kg

- 25 mg/kg

ai?i5-T=

-HOO mg/kg

-250 mg/kg
?100 mg/kg

mg/kg

Oral LD50

death in 3/3 in days

100 mg/kg or higher

- 2/u died in 9 days, 2 survived to luth da
- no deaths, all 3 survived to day in

1/1 died on day 2

- l/l died on day 3

l/H died on day 7, 3 survived to day 1?

no deaths, all survived to day 1?

for approx. 100 mg/kg; for 

Body weight decrease with larger doses of both 



and these animals developed anorexia, those
that died refused food towards end.



ataxia, stiffness in hind legs, myotonia

18817 

(3)





DOHZ

UBUQIB

gastrointestinal changes redness

in Email intestine, Some necrosis and inflammat'
hepatic necrosis, mild renal tubular degenerati-

deaths due

to nonspecific changes hepatic con?

many cases of death due to pneumonia.

Results of Chronic Studios:

 











m)

Mortality -

?2,5,10 mg or 
90?day test period

-20 mg/kg of compounds:

Body Height 
only in those animals that died

- survivbrs were free

all dogs survir

all dogs died during 3?

loss in weight occurred significe

of any 

both and dead, weakness, leg stiff.
difficulty in swallowing food, bleeding from gas

Blood Count - only change obser
in 3 animals that 

Organ Heights - no change in su

 

ved 

fall in c:
ied: 

rvivors; in 2 of

deaths, slight increase in heart and kidney
similar change in 2 of deaths.

Gross Pathology - dogs
ness in duodenum; 3
reddened duodenum and jejunum,
a generalized ieterus.

 

Microscopic Changes 5 cases 0
liver (not_related to dosages 
significance). Some duodenal 

infiltration of cells in the mucosa.
.serosa were normal.

Slight inc
in kidney sections in some dogs
and of doubtful significance).

showed areas of

one dog showed dif.

another demonstra

focal necrosis
or death, thus,
Yperemia, some 9'
Submucoce
rease in of ca;
(unrelated to c;

18518



- 

(H) Commeqt:

OF not as severe as those of in
acute studios.

?delayed dcath in chronic studies with may indicat
a cumulative effect of 

 

?signs observed in dying animals in acute study differs:
from those in chronic study.

(5) Summarv: acute oral LD in dogs 100 mg/kg, doses
50 .: -
f~ at this level or higher produced a definite myotonia with

anorexia and weight loss.

r. acute.oral LDSO in dogs 100 mg/kg or higher;
such doses produced only signs of a mild Spasticity.

[a Death during the repeated administration of 2,u-D or 2,b
was not related to pathological changes in the liver,
kidney, or other organs examined.

0.. "Summary of Toxicological Information on and 
Type Herbicides and an Evaluation of the Hazards to livestock
associated with their use?, (Rowe and Hymas, 195?).

(1) Early in 1950, Grigsby and Farwell published results of
experiments with and 2,u,5wT involving domestic

livestock, concluded "it seems that the farm use of these
materials for pasture weed control is a reasonably safe
procedure."

(2) Commercial Herbicid.s - formulations of active ingredients
appearing as a or an ester (soluble
in 011); most formulations also contain a dispersant, a
solvent, a wetting agent, perhaps a diluent (usually con?
sidered inert ingredients though they may affect herbicidal
effectiveness).

(3) 0ral_Administrati3n_of Sinfle Doses:

 

 

test materials administered primarily by intubation.

Or olive oil or corn oil, solutions used









(C) - 
rats 500 mg/kg 375 mg/kg
mice 389 mg/kg 368 mg/kg

guinea pigs 381 mg/kg H69 mg/kg
chicks 310 mg/kg 5H1 mg/kg
dogs 100 mg/kg 100 mg/kg

toxic of include loss of appetite,

weight loss, depression, roughness of coat, gereral
tenseness, muscular weakness; post mortem findings -
stomach irritation, minor liver and kidney injury,
lung congestion.

Oral Administration of Repeated Doses of 

 

Rats - fed 5 times/week by intubation, in olive
oil solutions. Dosages were 0.0, 3.0, 10.0, 30.0,
100.0, 300 mg/kg.

1n

Results no edverse effects ?t 1
mg/kg levels; l;g mg??g group - gastrointestinal
irritation, swelling in liver, depressed growth rate;
300 mg??g group failed.rapidly died, severe gastro-
irritation observed.

another Rat Study - fed for 113 days at levels
of 0, 100, 300, 1000, 3000 and 10,000 ppm.

1 ?n
J, .Lu, Ju

 

Results - no adverse effects in control, 100 and 300
groups; 1000 group suffered depressed growth
rate, excessive mortality, increased liver
weights, liver swelling; 3000 and 10,000 groups
were destroyed after 12 days since they were not
eating and were losing weight; examination revealed
increased liver and kidney weight, 1. and k. patho?
logical changes.

Chicks - fed for 7 days at levels of 0,500,
1,000 and 3000 ppm.

Results only adverse effects in 3000 group 

reduction in food intake and retarded growth_rate
(no histopathological exams made).

18.620-

U1

lI 

 

 



(5) Qg?l??dministration?te Steers (experiments performed
to see if extrapolation of data on small animals to larger
animals is Justified).

 

 

"Esteron Brusn_5iller? used (formulation of esters oi
and administrations made by intubatio'

 

?experiment 291 kg steer given single dose of
1000 mg/kg no toxic 

 

v-experiment 295 kg steer given 1000 mg/kg esters
for 3 days. 

results: general depresSion, decreased food and
water intake, decreased rumen motility; animal
died on 3rd day after last dose (death was undra
matic). Necropsy revealed dry rumen contents
with strong smell-of esteron, abomasum was im?
pacted, intestinal contents entirely fluid, con-
gested mcsenteric vessels, spleen dark and
shrunken.

?experiment 295 kg steer given 500 mg/kg for 2
days; 

 

results: on 3rd day, animal was off feed, rumen
motility ceased. On day, steer appeared
_perfectly normal, no discernible after?effects.

 

~experiment 336 kg steer given 500 mg/kg for 3
days. .

results: no toxic observed, animal
remained on full feed.

?experimcnt 250 kg steer given 100 mg/kg for
15 days. 

 

results: outward appearance normal. Killed
H8 after last dose for study. Duodenal
hemorrhage, abomasum irritation, hemorrhagic
necrosis of liver, fatty degeneration, slight
edema-and congestion in kidneys were found.

15821;

r? (6)









"u











"Karon" used in another experiment: 250 kg steer
given 100 mg/kg Karon for 15 days.

results: outward appearance normal. Killed NB
hours after last dose for examination. Very
slight edema in kidney, but this is frequently
observed in untreated animals; its significance
is questionable.

Discussion:

Dogs more susceptible to and than other
species studied; chicks appear tolerant.

 

"Inert" ingredients in commercial formulations have no
effect on toxicity of active materials.

and can be tolerated without adverse
effects in doses only smaller than those which
cause toxic effects when given only once - these mater-
ials have a low degree of chronicity, therefore.

There is a similarity in susceptibility of cattle and
small lab animals to these herbicides.

If esteron were applied at the normal rate of no more
than mg/sq. ft. (or 2 qts. per acre) a 350 kg

(770 lb.) animal would have to consume all forage

on sq. ft.to acquire a 100 mg/kg dose?of herbicide,
a level at which ill effects would not be eXpected.

. . there is little, if any, direct hazard to live?
stock or wild-life foraging areas treated with herbi-
cides of the type described herein. This has been con?
firmed by the extensive use of these materials over a
number of years without any proved cases of adverse
effects." -

Spraying of certain plants with can increase their
nitrite content to hazardous levels, but some of these
plants were sprayed accidentally, and others are not
ordinarily eaten by livestock. Toxic amounts of these
materials can be obtained if animals have access to
spray tanks or other containers of the material.

18822

DI

00H2060M8

"Results of Toxicological Tests on Dow, August

1950.



(2)

(3)



(5)

(data submitted to FDA Hearing on residue tolerances).

Because of similarities between and residue
tolerances for both should be same - 20 "would provide
a wide margin of safety.?

10% solution of in propylene glycol introduced
into rabbit caused pain and irritation immediately,
severe conjunctivitisand moderate corneal damage apparent
5 hours later, continued for 2 days before healing began.

10% solution of in butyl carbitol acetate applied
to rabbit ear and abdomen caused slight irritation.

is moderate in acute oral toxicity for the four
species, rat, mouse, cavie, and chick (rat more resistant,
chick most susceptible);

Handling hazards and precautions:

-swallowing, although unlikely, could cause serious ill
Effects; induce vomiting. 

-avoid contact with eyes and skin, don't wear clothing
or shoes contaminated with materiall

?no inhalation studies to date, but doubtful that toxic
concentrations would be inhaled due to irritation it
will produce in upper respiratory passages.

"Toxicity tests in NuAmine Report to Diamond Alkali
Company by Hazleton labs, February 1962.



(2)

Nufmlne 1,3 - propylene diamine esters of and
--

Acute Oral Administration NuAmine given to Sprague?
Dawicy rats (6 groups of 5 each) by stomach tube, either
in water emulsion or undiluted dosages of lO,3l.6, 100,
315, 1000 or 3160 micrograms/kg body weight. Concentra?
tions of material varied from it, 10%, or 100%. Observa-
tion period of-lN days following dosing.

 

C:
Pa
6%


$9
$0

 

4

'0be a

shortalitv: After 2? hrs., 2 in 1000 microgram/kg
group, and all in 3160 microgram/kg group died.
On day 12, another in 1000 microg?am group died, and
another on day These were only deaths recorded.

 

Other Effects:

appearance and behavior, increase in body weight, no
patholOgical findings.

and 100 micrograms/k: groups: normal

-316 seemed normal, 1 was depre.
on days lE?lH. Slight increase in average body weigh
2/5 showed congestion and hemorrhage of lungs.

~1000 normal at first, 1 showed
and labored reapiration after 
hrs. 2 died after 2? hrs., survivors showed depressio:
and labored resniration. After 48 hrs., all survivor:
appeared normal; at 2nd week, all showed unthriftines;
depression, labored respiration and ataxia. 2 more
died on days 
co.3e ion, kidney
and stomach hemorrhage, hematuria, intestinal inflame-
tion, discoloration and/or reduced size in kidneys
spleen and liver, stomach inflammation, congestion of
fat around tests.

- .- I, 1 
AutopSies shemed lurg ?drone- 

~3160 normal following
intubation; depression and ataxia after 1 hr.; pros?
tration, ptosis salivation, labored respiration, poor
reflexes after 5 hours; 1 death at 5 hours, survivors
showed hematuria. Total mortality by 2? hours.

 

Autopsios revealed congestion of lungs, adrenals,
liver and kidney and stomach mucosa hemorrhage
intestinal inflammation; bloody fluid in stomach and
urinary bladder.

(3) Acute Application NuAmine applied to eyes of



3 albino rabbits (left only, right served as control).

 

16524



(Ll)

UOHZUBUAZU

Single application of 0.05 ml of undiluted test material
made into conjunctival sac. Periodic observations for
1 week. Food (Purina rabbit pellets) and water ad lib.

Results: Animals seemed normal throughout study, slight
decrease in body weights in some. 

Eyes showed erythema, film over cornea, slight vas?
cularization of sclera and nictitating membrane, lacrima?
tion, edema of lids (obscuring and preventing thorougi
exam after a few hours), exudation, corneal opacity, and
corneal lesion (1 animal).

Acute Dermal Application - albino rabbits used; single
application of 50% volume/volume aqueous emulsion of
NuAmine at dose of 0.1 mg/kg body weight, and single
application of undiluted test material at doses of 0.1,
0.5, 2.0 ml/kg.- Test material applied to abdominal skin.
"Purina Rabbit Pellets? and water ad lib.

 

Results: 3 deaths at 2 ml/kg level, no deaths at any

 

other level. Estimated acute dermal L050 of NuAmine
for albino rabbits 1.2 ml/kg.

l. 50% emulsion: application(0.l ml/kg) produced no
signs of systemic toxicity, no pathological finding
2? hours after application - moderate erythema and
edema. At fourth and fifth days, erythema and eden
subsided, but animals showed slight or moderate
atonia and desquamation. By 12th day, almost all
Signs of irritation gone.

2. Undiluted:

-0.1 ml/kg - normal appearance and behavior.
Autopsy showed kidney congestion, mottled and
blanched?livers.

-0.5 ml/kg - normal appearance and behavior.

Autopsy showed kidney congestion, and granular appe;
ance of liver.

10

18825

(5)

 

~342mmjz?? - 3 deatls at this level, on Nth,
5th, and 7th days of observation. 2? hrs. 
application, 3 animals showed depression and laborc
respiration, seemed normal. Toxic signs gradu?
ally became more severe: depressed reflexes and
coordination, labored respiration, ptosis, ataxia,
coma phonation, salivation. Autopsies showed con?
gestion or hemorrhage of lungs and kidney; bloody
urine in bladder, bright yellow urine in another;
liver damage (blanching and discoloration) and
toughened liver surface; spongy brain tissue with
ekeessive fluid in skull; extremely congested or
hemorrhagic appearance of skull cap and bone marrow
One survivor was sacrificed; showed congestion of
kidneys. Subcutaneous edema, damage to all layers
of the skin.

ger??leffeots 2? hrs. after application,

skin at all dosage levels showed edema and blanched
areas surrounded by moderate erythema. In 3 animal
that died at highest dosage, dermal irritation was
relatively unchanged at time of death. Survivors
in all levels edema subsided after 3-5 days
scab formations or marked desquamation, atoni
coriaceousness, and fissuring developed to en
observation.



(1. I.) 

AP
UL

Repeated Dermal Applications - albino rabbits given 1 ml/k
NuAmine in varying (A, 12, 20 oz./gal.
aqueous emulsions) 5 days a week for weeks.

Results: each group showed moderate degree of dermal

irritation erythema, edema, atonia, desquamation.

Several in 20 oz./gal. group showed fissuring. One
animal in each of the test groups died, all showed
systemic toxicity depression, labored reSpiration,
Sprawling of limbs, weakness, weight loss. Horibund
animals also showed prostratlon, poor coordination,
depressed reflexes, clonic convulsions. Abnormal
blood and/or urine findings in each group, severe
liver, kidney and skin damage in and 20 oz./gal.
groups.

E-l ?MicrosCOpie examination of bone marrow sections" from
Nunmlne study, Hazleton Labs report to Diamond Alkali
Co., 12/16/65. - supplement to report of 2/23/62; tests on
bone marrow taken from rabbits receiving repeated dorms
applications of NuAmlne~D/T.

'll

. 

VF.



Results: No distinctive effect on bone marrow at

2 concentrations (H and 12 Higher con-
_centration (20 oz./gal.) group showed somewhat
atrophied adipose tissue. A possible effect - mega-
karyoeytes (in 20 oz./gal. group) underwent degen?
erative changes, more so than in controls.

"Results of 90?day Dietary Feeding Studies of Dowanol 978

(ester of in Rats", Dow, 11/27/61

(H
e)

(3)



Dowanol Q78 ester of is an agricultural herbicide
Tamper liquid)

experimental procedure: 5 groups of 20 rats were
maintained for 90 days on diets containing 
0.3, 0.1, 0.03, and 0.01% of the ester. Food (ground Fame
Laboratory Chow) and water ag_lib, -

Results;

 

 

0.03 and 0.01% levels: no evidence of adverse effects
increase in average weight.of spleen in males, increase
in body weight of females - believed due merely to chance.

 

0.3% level: growth retardation in males, liver and
kidney/body weight ratio increased in males. and live:
were large and light in color, some swelling and necrosis.
Some swelling in male kidneys; female kidneys appeared
greenish, some swelling and necrosis. Increase in serum
alkaline-phosphatase determinations in males.

0.1% level: livers (m and f) showed some swelling and
necrosis, females showed hypercellularity of glomerular
tuft, swelling of renal tubular_epithelium. Increase
in average male kidney weight:

 

Qg??l?ilg??: the Dowanol 97B ester of is moderate
in repeated oral toxicity in 90 day feedings. Levels of
0.03% and below were tolerated without evidence of adverse
effect. Male and female rats showed some injurious effects
(liver kidney-patholog\) at levels of 0.1% and above.

'Long?term dietary studies (2 years, with rats and
dogs) may be required ifester is found in significant
amounts as residue in food.

G. "The TexiCOIOgie Effects of Certain Fungicides and Herbicides
on Sheep and Cattle?, Palmer and Radeleff, 196?

(n

"In contrast to insecticides, organic fungicides and herbi?
cides (with only a few exceptions) have generally been
considered to provide comfortable margins of safety for
grazing livestock when the compounds are used according

to directions."

16827-.

 

 

(2)

(3)



Sheep and cattle maintained on grain concentrates with
hay supplements. 

Commercially available formulations used, as fluid .
dilutions or in gelatin capsules, number of doses varies.

Results:

(1)

(2)

a) 2

Fungicides 
a) lgantan?: sheep are regularly poisoned by doses of
250 mg./kg or higher; reduction to 50 or 100 mg/kg
permits animals to eliminate most of material without
being harmed. Cattle more tolerant of Captan than_'
sheep. Depression and anorexia,'tendency to stay away
from group were only toxic signs in sheep; in one
heifer, diarrhea and abortion observed. At necropsy,
hepatic and renal changes noted but not remarkable.

b) ?gingb": Single doses of 500 mg/kg. tolerated by sheep
and cattle. 90 daily doses of 100 or 250 mg./kg. did
not poison sheep. In single poisoned sheep (15 doses
of 500 anorexia and bright yellow diarrhea
observed. Necropsy showed congestion-of the lungs,
hepatitis, and nephritis.

c) "Ceresan Sheep fed 1, 1.5, or 2 lbs. of treated
grain per day. Lethal to all, at doses of lb./day 
33 doses, 1.5 lb./day 22 doses, or 2 lb./day 6
doses. Signs of poisoning included anorexia, diarrhea,
weight loss, nasal discharge, stilted gait and lameness.
Necropsy showed lesions characteristic for mercurial
poisoning. (Ceresan contains mercury).

Herbicides 

administered as alkanolamine salt and as prepyle:
glycol butyl ether ester to sheep and cattle.' Very
little difference in toxicity of 2 derivatives. ?Remark-
able tolerance" of ?81 daily doses of 100 mg./kg. by
sheep indicates little probability of acute poisoning

in normal applications of the compounds. Toxic signs

in sheep - anorexia, depr=ssion, weight loss; in cattle 



muscular weakness, ataxia, dry, cracked muzzle, ulcera? 
tion of nasal mucous membrane. Necropsy - lesions varies

usually liver and kidney degeneratiOn;rumen stasis with
bright, undig.nted ingesta, hemorrhages in heart and_
excess of pericardial fluid.

13
166328

 


I

b)

c)

(3)



??mm'ddwid?oeza

anhnir:lsia:rc<l as s:1lt amid aLs
propylene glycol hutyl ether enter to sheep and
cattle. Salt appeared least toxic (sheep tolerated
?Bl doses of 100 mg./kg.) while esters were lethal
at 369 doses of 100 mg./kg. Enters were lethal to
sheep and cattle after 7 doses of 250 mg./kg. 

2, H, somewhat more toxic to sheep and cattle
than 2, "although poisoning under ordinary
circumstances of use would be most unlikely."

(2 - (2, N, S?trichlorOphenoxy) prepionic
acid) administered as butyl ether esters of propionic
acid derivative to sheep and cattle. Markedly more
toxic than or 2, u, (sheep succumbed to ll
doses of 100 mg./hg., cattle to 29 such doses). Signs
of poisoning: anorexia progressive weight loss. 
Necropsy showed degeneration of liver and kidneys,
hemorrhages on hear surface, congestion of visceral
blood vessels, swelling of glands, rumen atony
with large quantities of ingesta.

 

Benzoic Acid Compounds - dimethylamine salts of 2, 3,
6?trichloro~ (iiysben) and 2-methoxy - 3,6-dichloro ensoic
acids (Banvel D) tested. Both compounds more
toxic than phenOxyacetic acids studies. at dosage
of 250 mg./kg. 10 doses was lethal to the one sheep,

but the cow was unaffected; at 500 mg./kg. 2 doses, the
compound produced mild poisoning in one other cow.

Eanvel in doses of 250 mg./kg. 10 doses did not affect
a Sheep, but killed another sheep at 500 mg./kg. 2
doses. A cow was not affected by 5 doses of 250mg/kg.
Signs of poisoning: salivation, trembling, depression,
tympanites. Necropsy showed congested lungs, hemorrhages
on heart, pericardium and larger blood vessels, undigested

feed in rumen, edema in respiratory system, inflamed kidnet

liver engorged with blood.

Triazines - (Propazine, Simazine, and Atrazine): admin?
istered to sheep_and cattle; Atrazine was lethal at all
doses tested (50 mg./kg. 199 doses, 100 mg. 16, 250

2) for sheep, and lethal to the cow tested at 250 mg./kg.

2 doses. At the 250 mg./kg. level in sheep, Simasine
was more toxic than Prepazine; at 50 mg. kg. Simasing


was more toxic than ?trlging.
anorexia and depression.
and Simazine): muscular spasms and fasciculation, stiff
gait, increased respiratory rates. Necropsy in sheep

and cattle given Atrasine showed degeneration and dis?
coloration of the adrenal glands, congestion of lungs,
liver and kidneys, presence of undigested food in rumens.

v: 16829

I.

Signs of poisoning (.ropasin



DOHZUSUHS

Liisec:lljnaeolu; 

 

 

le?pon (2, 2-dichlor0pr0pionic acid) sodium

-salt is of a low order of toxicity (500 mg./kg. 
10 doses not toxic to sheep or cattle, one sheep
tolerated H81 doses of 100 mg./kg.) No poisonous
signs. .

Hg?_?mi?p (2-methyl (H?chloro?o-toloxyacetic acid)
alkanolamine salts poisoned one sheep lethally at
383 doses of 100 mg./kg.; a cow was poisoned by 8
doses of 500 mg./kg. but survived. Signs of poison?
ing in cattle: anorexia, chronic tympanites; in
sheep, anorexia, depression and ataxia before death.

Fenurg? (3?phenol?l, l?dimethyl urea) - appeared more
toxic than most herbicides discussed, but high
dosages still required to produce poisoning 
250-500 mg./kg. 2?10 doses). Toxic signs: de?
pression, anorexia, complete incoordination (heifer 
abortion), slow recovery. Necropsy: lung congestion,
'heart hemorrhages.

Bandane pentadiene isomers) - sheep
poisoned by'3 doses of 250 mg./kg. but survived,

while same dosage was lethal for cattle. Toxic signs
in sheep: salivation, anorexia, tympanites, 
and depression. Toxic signs in cattle: tense muscles,
muscular spasms. (Even when dosing stopped, signs
progressed, duplicating tetanus). Death preceded

by convulsions. Necropsy: hemorrhages of cerebrum
'and medulla oblongata, hemorrhage of right psoas
muscle, skeletal fat replaced by gelatinous material,
lung congestion.

Discussion: studies offer no "reason to alter the
already generally accepted concept of the hazards pre?
sented by these herbicides." Further study needed Lg
toxic signs.

Conclusions: feeding Ceresan (13?38 mg./kg.) on
treated grain by mistake can easily poison sheep;
other materials studied are safe when used as recom?
mended.

"Commercial Fishery Investigations", Philip A. Butler Acute
and Chronic Toxicity Studies,

1961:

purpose of project: to learn how to protect and pre?
serve marine environment from possibly adverse_effects
of agricultural chemicals; discovery of pesticides
that may be useful in improving fish harvests

18830

 

(2)

(3)



(6)

(7)

(8)

(9)

(10)

  

"The need for this research does not imply that the
of pesticide formulations automatically
constitutes a serious threat to marine life."

the moat ha~mless of chemicals, including water,
is tox1c when present in sufficient quantity.?

different forms of marine life may react quite dif?
ferently to the same chemical

four bioassay groups (phytoplankton metabolism, oyster
shell growth, shrimp survival, fish survival) tested
for sensitivity to four pesticides methyl
Parathion, phUsdrin, phosphamidon). Phosdrin and
phosphamidon Produced the least reaction; dyrene had
some effect on all groups, especially fish survival;
shrimp survival was very sensitive to methyl parathion.

phytODlanktOH microscopic sea plants, essential

link in conversion of solar energy into food for more
complex marinv animals. Using decrease in growth,

or decrease in carbon fixation as a standard, toxicitie:
of pesticides may be compared.

crustacea - shrimp is most valuable fishery; most_

PestiCides kill terrestial members of shrimp's
Phylum Some pesticides paralyze shrimp

and other crustaceans, rather than kill; toxic cri?
teria for this group is effective concentration causing
paralysis or death to half the samples within a stated
period (E050)

mollusks clams and oysters store high concentrations
of chemicals that exist in only trace amounts in the
surrounding sea water. Mollusks, however, are able

to Close their valves and protect themselves from toxic
substances in the environment. Thus, tests must be
conducted at nhhlethal concentrations; growth is an
Objective indrx to measure, using ECSO measure of effect

fiSh.- YOUHBPP specimens usually most quickly affected
by pesticides.

2; "a 5:2 butyl ether ester):

ECFJ for oyuivr shell growth 0.1? mg./litCr (96 hrs-
1 wee recovery period.



"Commercial Fi??g?ivu Investigations," Philip A. Butler 

Laboratory Studies HUN Toiicology

?16 . iss31

(1)

(2)

'UaHZb?uuzz

testcnitniused
significant decrease in productivity at 1 ppm, al?

'though at lower concentrations an increase in pro?

ductivity rate was frequently observed (probably
because pesticide was toxic to animal part of natural
phytoplankton community). Herbicides and DDT very
toxic to some plant species.

crustacea post?larvae of brown shrimp and blue
crabs are quite sensitive to chlorinated hydrocarbon
insecticides. All shrimp and most crab tests con?
ducted in sea water with acetone stock solution of
pesticide (constant flow systems, in which test solu?
tions are renewed continually, have advantage of in?
suring sufficient oxygen and desired concentration

of pesticide). After exposure to lethal or near
lethal concentrations, crabs and shrimps often mori-
bund for hours. 

 

mollusks oyster and clam tested, constant flow

system with acetone?stock solutions of pesticides.
Oyster closes its shell when sufficiently irritated

by contaminated water does not feed while closed and
therefore does not grow, therefore relative chemical
toxicity is indicated by changes in growth rate (shell
growth). Controls 
inated hydrocarbon insecticides were most toxic pesti?
cides to oysters - most inhibited shell growth at

well below 1 ppm; other pesticides 
and carbamate insecticides) had little or no effect
on oyster growth - they decomposed in water. Meta?
bolism increased at summer'temperatures, therefore,
at higher temperatures some chlorinated hydrocarbon
insecticides killed oysters at concentrations which
at lower winter temperatures only stopped growth.
week recovery period for reduced growth rate; data
indicates that "a Single field application which might
temporarily affect oyster growth would probably not
have a lasting effect." Chronic, rather than acute,
exposure to pollutants is more likely.

*n ~hnv.r4-L\ 
  4-?va 

1?4

Chemicals tested included aldrin, dieldrin, DDT, toxa-
phene, malathion and acetone (acetone used as a solvent
in program, so tested for possible toxicity). In
general, no decrease in growth rate; treated groups
sometimes grew better than controls. Oysters
did not acquire tolerance to pesticides.

430,-

)0 for decrease in shell growth:

acid (temps. of 16 
at 2 
acid (temps:

3000) no decrease

- no decrease at 2 p;

17 1883??

 
1'

"funk
n-H

 

(5)

J.

"Commercial Fishery Investigations", P.A.
Toxicity,





gish juvenile white mullet longnose killifish
Used as test animals in running; sea water aquaria.
2H and N8 hr. median tolerated limit values 
ottaincd. Chronic low?level pollution studies im-
portant, since many marine fish pass their early
growth stages in estuaries.

3 groups of Spot (leiostomus xanthurus) exposed 3
mos. to sublethal concentrations of dieldrin (0.1,
0.01, 0.001 ppb), in running sea water. High
mortalities, 30?37%, but also in controls. No dif?
ferences in mean though some experimental
fish had axial skeletal distortions, not in controls.

Survivors exposed to 2 dieldrin to determine

if earlier low?level exposure had created any
resistance: 80% of experimental fish survived Eu?hr.
exposure, while previously unexposed fish all died.

acid tested on longnose killifish showed
no effect at 50 (2H and H8 hrs. with
19? water temperature.

 

?Esteron 9Q (formulation): for 50% mortality
in mullet 1.5 (2H and H8 hrs.) at in
killifish.= 3.5 hrs.) and 3.0 (M8 hrs.
at 

Future considerations: to determine which pesticid:
now considered necessary may be expected to cause the
least damage to marine resources. careful selection
of the control agent and method of application can
lessen the current pesticide hazards."

 

Butler Acute
19b5

Crustagga - shrimp - most valuable commercial fishery
resource, spend part of life span in estuaries sus-
ceptible to pesticide pollution. Since they are
rather closely related to insects, insecticides are
particularly toxic to them either kill quickly
or paralyze them so they lose sense of balance.
conducted N8 hrs. in flowing seawater.

Tests

Egsults: had no effect on brown shrimp
mortality or equilibriam at 1 after ?8 hrs.
(an. water temp. 

hutyl ether ester tested
on brown shrimp in water caused 10; mortality
(or loss of equilibrium) at 1 after 2? hrs., and
20% mortality at 1 after ?8 hrs.

 

(2)

tested w: adln; in 
water had no effect on Lgi-ium
after 2? hr. and H8 hr. esposurc at i levei.

El?n younger specimens most quickly affected,
therefore juveniles used in tests. Hun water used,

Results:

Eglx?lycolm?g?vl ether ester tested

on Spot in ILUC water ?8 hr.
E050 of 0.32 for mortality.

Zydon also tested on Spot, in had
no effect after 2? and U8?hr. concentrations
at 1 ppm.

K. "Recommendations for Minimizing Dangers of Pest Control and
Pesticides to Fish and Wildlife?, John L. George June, 1963

 

(sure there is real need for pesticide use

00

discuss possible hazards with biologists prior to use:

avoid treatment of any significant portion of range
of a rare species or one with specialised habits -
consult with conservation officials -



treat minimum necessary area; avoid direct treatmentco
of streams, etc., consider danger of runoff contam_
ination

judge danger of chemical both by its toxicity and its
disappearance rate 

use method of application which will minimize contact
with wildlife. Emulsions are more toxic to water environ
ments than oil solutions or suspensions. Granules

often concentrate the effect of the toxicant in one
stratum.

no double-dosing
avoid main Spring migration, nesting pvviods of birds,
etc. when choosing time of treatment consider possi?
bility of using repellants or scare devic?3 to move

mobile wildlife from area. 

physical or biological control methods have advantage
over chemical methods - more Specific. reduce pests
without loss of beneficial population.

susceptibility to different pesticides varies greatly
among animals

19 18834

v- -- --



 

(ll) DDT susceptibility: 0.1 5 lbs./acre 
heavy kill effect on crustaceans and fish,
moderate kill to amphibians and reptiles,
some kill to birds, no effect on mammals until
5 lbs./acre level

table with toxicities of other chemicals as compared

(12)
to DDT: derivatives 

rats - LD50 of (single oral dose)

bobwhites - 0.2 chronic toxicity LD50 (10-100 daysLDSO

pheasants




mallards

bluegills LD50 of 0.1 (96 hr. test)

(13) disposal of pesticides always difficult procedure -
unOpened containers should be returned to wholesaler
or sold; disposal of opened containers:

burned in a flame (liquid organic chemicals)
addition of lime (wettable powders)

dumping into sanitary landfill-type dump, contain-
erized if soluble

empty containers burned or dumped - do not
clean equipment at creeks or waterways

.-

OEUUQUZHDU

 

 

17. How, "The Analysis of Rice for Residues of 5, 
il?utr-edients of DowJ 24, ll, 9min? Wm] Ir-an.
I (2) directions for use - weeks after emergence of rice
(7?9 weeks when flooded), treat with - 2.5 pts. product
I in approx. 5-7 gallons water per acre 0.5?1.25 lbs.
2, u, S-T acid)

f? - (3) with some weeds, up to 3 pts./acre may be necessary, but
yield may be reduced

(N) treatment after flooding is safer than treatment before
flooding 

(5) warning: do not apply to vegetables, flowers, grapes,
fruit trees, ornamentals, cotton or other desirable
plants which are sensitive to 2, U, do not permit
spray mist to drift since even minute quantities of the
spray may cause injury_tolthem.

 

(6) residues methods for determination extraction,
chromatography, colorimetric determination, standard
I curve. Method capable of determining residues at
level of 0.1 

results indicate residues less than 0.1 when applied

at levels of 0.75 and 1.25 lbs./acre.
(2 locations: Beaumont, Texas and South Dos Palos, Texas

levogazuon

2i 15635

 



l8. j?riige, 
liepord; on 511 Rice atulftiee Stiiuv"

A. ?urposg: to determine residues of in rough
rice and rice straw from Arkansas, Louisiana and Texas

(1) no detectable residues found in any rough rice sample

(2) Vgogisiana straw samples contained 1.0? residue
after 0.75 and 1.5 1b./acre treatment; 0.85 found
after a single 1.5 lb./acre treatment

(3) Texas straw contained 0.18 after double treatment
.(0.75 and 1.5 no detectable residue after
single treatment(l.5 lb/acre) 

(H) Arkansas straw detectable residues of 0.02 
only after single treatment (this level
residue is close to limit of detection and may be due
to background interferences)

(5) no detectable residues of "bound" found (both
double and single treatment) in rough rice from all
3 states

(6) 1.5 lb mistakenly applied over a Silvex
treatment; analysis revealed 0.02 

(7) average recovery of from rice and straw was
Bu:10.5%

B. Supplements, etc.

 

.procedure for free chlorOphenoxy acids
(2) procedure for bound chlorophenoxy acids
(3) appendix:

Arkansas - rice seeded 5/6/69, treated 6/16 and 7/11
harvested 10/3u6 

Louisiana - rice seeded 5/12/69, treated 6/19 and 7/
harvested 8/28

Texas rice emerged 5/8/69, treated 5/28 and 6/23,
harvested 8/15

C. Letter From Hoodhury to Dunn (both Hercules, Inc.) 6/3/69

re coeperators rice residue project, samples
they are using and approximate harvest dates. 

22 15637

 (ll.i 

E.

 

Qbiegtjye: to obtain samples of rough rice, straw,
mill rice, rice halls and rice polish and bran of
adequate size for residue analyses data to he used
in supplementing; a petition for establishing
tolerances for 2, silvex and MCPA in or
on rice and milled fractions thereof.

general consideration: conditions for growing should
conform to those typical of the area

 

eevoguznoc

rice variety: should be typical of those used in the
state.

formulations: amine salts of low
volatile ester of silvex, and MCPA to be used - this-
form is most widely used and recommended

rates and timing of application: highest registered rates
of application and shortest interval between applications
and harvest consistent-with registration limitations
should be used, to obtain highest residues. Single
treatment, unless two applications are required.

 

Suggested treatments:

 

b-T 0.75, followed by 1.5 lb./acre, 3?5
weeks after emergence

(2) 2, N, - 1.5 lb./acre, just before panicle
initiation (when internode length is 1/2 in. or
less)

size of plot: 4' 20' minimum, replicated times, is

suggested

rice products for analyses: about 10 lbs. straw and
20 lbs. rough (unmilled) rice from each treatment will

be required.
Rice - USDA registered uses:


1.25 lb./acre tiller to boot and before flooding
weeks after rice 

1.5 lb./acre apply after flooding (2-3 weeks)
or 7?10 weeks after planting

.23

 

f?u



K.

Rice "Produc ion and Acreage

Arkansas ?30,061 acres (2h.89? of US production)
California 323,630 acres (2H.12Z of US production)
Louisiana 512,88u (2H.26z of us production)

Texas ?64,732 (23.99% of US production)



Average US yield/acre 3,553 lb (5 year average)

Letter from Hoodburl (Hercules) to Baker (Louisiana State
Univ.), 3/11/ 9 re: rice residue project, enclosing copy
of suggested protocol.

Letters also sent to:

-Flinehum (Texas A Univ., Beaumont, Texas)
-Smith (USDA Rice Branch Experiment Station, Stuttgart,
Arkansas)

Letter from Flinehum to Devine, ll/lH/69, notifying of

shipment of samples from project and listing treatments.
Treatment (1.5 lb/Acre Silvex) mistakenly applied over
Treatment (0.75 1.5 lb/Acre Silvex). Each treatment

- replicated times.

Letter from ?mith to Hoodbury, 11/25/69, notifying of
shipment of samples from project and listing treatments.

Letter from Baker to Woodhuri, ll/ZM/69, notifying of
shipment of samples from project and listing treatments.

 

Letter from Elipehum to Hoodhury, 2/5/70, describing pro-
ject at Beaumont.

m: 16539

"Other pesticides sod besides Silver

I
and MCPA:
Arasan, Aldrin; propanil

Yield Data:

-2,u,5?T (0.75 1.5)
(1.5)

no herbicide

2830 lb/acre (lowest in grour

3031 lb/aere
3579 lb/acre (highest in grO'

ll

QEVUBUZHUU

 

188430

. 25

19.

Dow, effect of irradiation upon TCDD 5/95/70:

?l.02 mg TODD, in lOOznl.ehloroform saturated with water,
exposed for 2?1/2 to UV light (GE sunlamp) at 

?50 to 100% debraded into other compounds (not identified).

"a dramatic change"

BEvogozngg

26

20.

1393-: .(12/8/611, "Solum 111.3; of in various

at 

?least soluble in watdr
?mos soluble in O?dichlorobenhnne

27

ZEVUQDZHOU

21.

USDA.Notice, NRC Pesticide Residues Committee



"Statement

for Implementation of Report on No Residue and Zero
Tolerance? (approved by Orville Freeman, Sec'y, 



(2)

(3)

(N)

11 recommendations of NRC committee, from June 1965
study and subsequent reporti

"No residue? and "zero tolerance" concepts are
scientifically and administratively untenable
and should be abandoned.

"Negligible residue" or "permissible residue?
registration suggested. 

Registration for residue-producing pesticides
only if negligible residue in line with FDA
regulations.

"Negligible residue? figure should be published,
as well as analytical method for determining
residues.

If pesticide is known to be too hazardous for

H114

.
. ni11nvu Ht.- 4 '0
3. 3. 1 budshould be refused.

Responsibility for registration properly rests
with USDA.

Schedule for transition to new procedure to be
decided by USDA and HEW.

Manuals, mutual assistance encouraged.

(1) Formal program for education in residue analysis
is needed - agencies should cooperatively
sponsor program.

(J) Expanded research program needed in this area.
After considering report, Agricultural Research
Service and FDA agreed on certain general principles
and procedures.

Federal Food, Drug and Cosmetic Act: any pesticide
in or on food shall be deemed unsafe unless a
"tolerance" has been prescribed. 

Both agencies agree in establishing finite tolerances
concept. 

lasis_



(7)

(8)

(9)

(10)
(ii)

(12)

(13)


us)

(16)

[aesticyides shall. not be registered
under FIFHA unless a finite residue level is pro-
vided for by Act.

"Tolerances should be established on the basis of
data in petitions presented by proponents to
establish that such uses will be safe".

ZHOU

"Agricultural uses of pesticides for which it can :3
be concluded there is no reasonable expectation ofCD
any residues on the food will be considered as nonCD
food uses and can continue to be registered in the"
of a finite tolerance" (until or unless
. e0
reSidues are discovered later on).
For tolerance purposes, chronic feeding studies in
2 species of animals and reproduction studies re-
quired (unless only negligible levels are involved,
when 90 day feeding studies on 2 species may be
sufficient to provisional tolerance).

"No procedure or formula is to be employed which will
serve to override scientific judgement based on ade-
quate safety data". 

"Prompt action will be taken on petitions for toler-
ances.for negligible residues of such pesticides?.

All petitions should supply an analytical method.

"It is reasonable to expect that uses of persistent
pesticides on crops or in soil in which crops to be

4. ll

grown may result in residues on the crop at harvest

"Zero tolerance? or "no tolerance" registrations
should be discontinued as of 12/31/67 unless finite
tolerances established or in process of being es?
tablished; in no case should former registrations
extend past 12/31/70.

Both agencies

(FDA and USDA) can receive ans process
petitions. 

Comparable problem with inadvertent and unavoidable
residues which needs resolution.

Public Health Service of HEW is available for consul?

tation and correlating human experience with animal
experience.

ld?di

re

22.

A

Statement by Julius E. Johnson (Dow) before Senate

Subcommittee on Energy, National Resources, and the

Environment, 

(1) 10/29/69 announcement by Dr. Dubridge (Office
of Science and TechnologY) referring to birth
defects observed in Bionetics test.
ceded final report of Panel on Teratology to
Mrak Commission (Commission on Pesticides and
their Relationship to Environment and Health,
appointed 5/69 by Secretary of HEW).

This pre?

members had not seen Bionetics report at that

time.

(2) sample in Bionetics tests came from
Diamond Alkali Co. (no-longer makes 



Commission

2,?,5?trichlorophenol came from Coleman Mathison

Bell, via McKesson Robbins, via Dow. Quality

of 2,?,5eT is related to quality of its inter-

mediate 2,U,5-trichlorophenol; possibility of

a highly toxic impurity being formed in 

trichlorophenol as a side reaction under con?
ditions of elevated processing temperatures.?
Most sensitive toxic reaction observed in

humans - chloracne, mostly prevalent on face,
neck and back.

Impurity not formed during manu-

facture of Dow from the triehlorophenol,

nor does it form on storage even at high tem-
peratures. Impurities concentrated in waste
oils from 2,4,5?trichlorophenol process.

(3) Early control test - rabbit ear test for
dermatitis.

(M) 196? Some Dow workmen developed chloracne;

rabbit ear test showed chloracne potential of

waste oil from process
was building up to a danger point, caused by
operating changes made to improve production
capacity. Waste is routinely destroyed by
incineration at high temperature. ExpoSure
to waste oil caused acne in workmen. Plant
shut down; it was discovered that principal
offending impurity was TCDD (found by gas
chromatOgraphy). Plant was redesigned to-
ensure minimal TCDD production. Since

30

Kb


CRI-

(5)

(6)

(7)

(8)

(9)

(10)



1966 Dow and have
met specification of less than 1 (most were
less than 0.5 ppm) TCDD.

In Bionctics test, showed
no significant increase of anomalies, but 
sample did display a significant increase of
anomalies. Samples of Diamond Alkali 
contained tolevels of 16 ppm, according
to records.

Above information presented to Mrak CommissiOn on

11/7/69-

Low mamm~lian and absence of
reports of increased birth defects in cattle or

sheep grazing treated ranges caused Dow

to-doubt Bionetics study's reliability.

Tests performed at National Institute of Environ?
mental Health Science (NIEHS) labs at Research
Triangle, North Carolina. Previously agreed to
try to duplicate Bionetics study, using Sprague?
Dowley rats (superior to mice tests) and using
Dew of regular production grade. Dow
would provide samples of and TCDD to
NIEHS labs.

On 12/11/69, Dr. Courtney (NIEHS) provided sample
of used by Bionetics labs; examined at
Dow and showed to cause positive rabbit ear re-
action characteristic of the contaminant, and gas
liquid chromatography indicated presence of 27:

8 TCDD.

l/l2/70'report to HEW showed that Dow of
regular production grade-did not cause birth
defects in rats as determined by gross examina-
tion of fetuses. Dosage levels used were selected

'in consultation with Drs. Falk, Courtney and Gayle

of NIEHS. Pilot study with pregnant rabbits also
showed had not caused birth defects. Later

l?843
31 a

as

(11)

(12)

(13)



(15)

DUHZUBUMZ

microscopic examination of ssues and skeltons
confirmed preliminary findings of no causality.

TCDD teratology study in rats showed high level
of maternal and fetal toxicity; concluded that
presence of TCDD in Bionetics sample could well
have accounted for observations reported and at?
tributed to 

Dow physicians have made in-depth evaluation of
health of 130 male employees exposed to 
in manufacturing operations for from 6 months-to
approximately 20 years; 50 clinical parameters
evaluated; control population ?,600 individuals
for whom similar data were available. Conclusion:
no evidence of adverse effects.

Dow's belief 2,?,5nT with less than 1 
TCDD presents no practical hazard when used in
accordance with good agricultural practices.

Industry Task Force on Phenoxy Herbicide Toler?
ances established by National Agricultural Chemica
Association, pooled information, submitted petitio
for negligible residue tolerances of in
Dec. '67; in April of1963, FDA said more informa?
tion needed on metabolism and residues.

originally registered under "no residue?
tolerance; on h/l3/66 Secy?s of USDA and HEW
changed policy to finite residue tolerances. Sincc
Task Force data would not be ready before Jan. 1,
1970 deadline for new registration, the Task Force
withdrew its petition without prejudice to future
filing,wand requested extension of regis?
trations since no imminent hazard from continued
use was anticipated. In response, USDA extended
registrations to 1/1/71; Industry Task Force to
refile before 1/1/71.

32 168%?








.





"The Response of Rabbit Skin to Compounds Reported
to Have Caused Acneform Dermatitis". Adams, Irish,
Spencer and Rowe (Dow), from Jan. issue of

- Industrial Medicine;

 



(2)

(3)

(H)

Materials have been applied to inner surface of
the ear of albino rabbits and to the shaven belly
both undiluted materials and solutions of various
concentrations in olive oil, paraffin oil U.S.P.,
propylene glycol, ethanol and water. Applications
made once a day, 5 days a week, for weeks or
until a marked reaction resulted.

Some of strongest irritants produce a rapid des?
truction of the tissue (necrosis) without the skin
having an opportunity to show an active response;
milder irritants have some effect upon the tissues
Most irritants resulted in responses which tend

to develop rapidly and to subside in a short time;
response may include hyperemia, congestion, infls:
mation, exfoliation, edema, blistering, slbughing,
exudation, crustation, necrosis, induration, hair
loss. One type of response has a latent reaction
and a much more prolonged course - epithelial
hyperplasia, with its resultant thickening of the
skin, follicle enlargement and sequellae (varying
degrees possible, from slight to severe).

Indications of a relationship between reaction ob?
served in rabbit and acneform dermatitis of man: re
action in rabbit produced by 5 types of Substances
known to cause an acneform dematitis in man
(chlorinated diphenyls, chlorinated naphthalenes,
chlorinated diphenyloxides, crude chlorinated
phenols, and petroleum oils).

Other reports: epithelial hyperplasia-from various
oils, fats, paraffin, dyes; eczema from aniline

and coal tar exposure (also warts and acneform
dermatitis (chloracne)).

33


tats

IN



IO

UOHZUBMM

(5) Chloracne visible response of skin to irritant,

taking form of epithelial changes, (2) inflam
matory and degenerative changes, and (3) regenera
tive processes, (in order of appearance).

Dow: "Determination of (TCDD) in by gas?
liquid Chromatography" 6/22/65 Analytical Method.

(1) Dioxin can be detected at 1 level with a lower
limit of 0.5 possible at optimum operation
conditions.

(2) Accuracy of this method i 5% or less.

Abstract from "Teratogenic Study of in the
Rat", Emerson, Thompson, Gerbig and Robinson, Dow
(presented 3/17/70).

(1) Study to determine embryotoxicity or teratogenicit
of containing less than 1 TCDD, tested
on Sprague-Dawley rats. 

(2) 5 treatments groups, 25 females each

(3) Given 1,3,6,12 or 2H mg/kg/day of compound via
gavage in 0.25% METHOCEL on days 6?15 of gestation
Control group (50 females) received vehicle alone.

No clinical or gross pathologic signs of adverse
chemical effect were observed in treated dams; no
teratogenie or embryotoxic effects.

Results of study failed to substantiate Bionetics
Study.

Abstract from "Teratogenic Study of (TCDD) in the Rat",
Sparschu, Dunn, and Rowe, Dow (presented 3/17/70).

(1) Study to determine if TCDD impurity could account
for fetal abnormalities in test animals in Bionetie:
tests.

. 
18MB

r?I-

(2)

(3)

(N)

(5)

(6)

(7)

(8)

(9)

00112080445-

Dioxin given by maVaae in 9:1 corn oil -

. acetone solution in doses of 0(control), 0.03,

0.125, 0.5, 2.0, and 8.0 micrograms/kg/day to
groups of 20 (control) and 12 (treatment) preg?
nant Spraguc-Dawley rats on days 6?15 of gostatie
dams sacrificed on day 20.

No differences in fetuses from control and 0.03
micrograms group.

0.125 microgram group Shemed slight decrease in
average weight and intestinal hemorrhage (18/127)
and subcutaneous edema (22%80) in fetuses.

0.5 microgram level number of fetuses reduced,

number of resorptions and fetal deaths increased;
average'weight of viable fetuses decreasc
intestinal hemorrha33 (36/99) and subcutaneous ede

(31/65).

2.0 microgram level - only 7 viable fetuses, from
of 11 litters, numerous resorptions, intestinal
hemorrhage in of the 7; subcutaneous edema in u;
fetus had kinked tail and 2 misshapen feet.

8.0 microgram level toxic?to dams; no viable
fetuses; all resorptions were early and no fetal

.tissue found.

Delayed ossification of some sternebrae and skull
bones occurred throughout various groups, includ-
ing controls - not considered to be of practical

significance.

Results indicate high level of maternal and fetal
toxicity to be associated with could account
for Bionetics observations attributed to 


a
our
CD

 

 

?23.

DOHZOEOMB

USDA notice to Manufacturers, Formulators, Distributors,
.and

'no

Dec.

(1)
(2)

(3)

Registrants of Economic Poisons re "extension of cert
residue' and 'zcro toleranco' registrations beyond
31, 1969" (dated April 9, 1970).

Lists were extended until Jan. 1, 1971.

2,u,5?T uses on McIntosh apples, low bush blue-
berries, grains (cereal?undesignated), pastures
(grasses), rangeland clearances, rice and sugar-'
cane - extended by this notice;

Signed by Harold G. Alford, Ass't. Dir. for
Registration.

 

 

I-





2M. Specification for 2/27/69,
released For use by USDA etc. by G. E. Lynn.

Lists TCDD to be at a non-detcuLnb19 level, whether

produced by Dow itself or purchased by Dow for
resale, etc. 

37 . . 5:  



85:

a-



 
-. "7  5'34 I
:1 

'wdd 3931 sq on ?com SHOWS 1

?nL-s?u?a JOJ UOEJUDTJIOOGS notes" '98

infor quality testing /5/65. 01. assay and poir
 27.

 

MnLhod for detormin?tion of TCDD in
by gas?liquid Chromatography", 6/22/65.

 




:war

END OF VOLUME TWO

H0

osvogozmou

1E3

2680



Hr. ntnh-ralaml.

ll simply math-r ol' the t':ll'l' ancl Sophislit'ation of tho mothotl,
tla- thou to gut. tho. joh door. have built, Dr. ltaylcv has
out, an isolation lalmratory?l say hailt, I change that. to
t-quima-(l an isolation lalmralory We haro tho scit-nliata. We are,
In fart. rt'atly to pron-vial. The How. tht't'ol'oro, wonhl ho the time
to (l0 the analyses and verify thmn.

Mr. lhc-awrr. Thank you rvry nun-h.

Ht-nator (h'ntlonu-n. thank you. It has been an interesting
and inl'orinatiro morning. Congratulations again for tho. effort. that.
you. I am aura, put; into ?hm-lopng and than persuading dopart-
mental awrptanyo of tho soggrattal I hope improve-
na-nt. that. has?: law soon Will he written.

Dr. Hana-11'. Thank you, Mr. Chairman.

Hanator llama \Vo adjourn. to rt-alnno tomorrow at. 11 nan. in
tho morning in the. hearing; room of the on Colman-co 5110.

("ll la-rcnpon, at. 1240 I).tll., the hearing.r recessed to at.
11 nan. on Thursday, Juno 18,1970.)



 

 

 



EFFECTS OF RELATED 0N
MAN AND THE ENVIRONMENT

 

THURSDAY, JUNE 13, 1970


(Iota n'rnn on (lost .n ma ac,
Sont'oMMI'rn-zn l'iNl-IltnY, lh-zaoom'r-za,
AND 'rmr. 
ll?us/aing?an, 

'l'ho nwt. lmranant. to :uljournmont, am.
in room ?lm, Now Ht-nalo llnihling, llon. Philip A. llart.
(t-hairman of NW [in-Hiding.

l?rvHI-ot St-nalol? llarl.

Sc-nalor ll 'l'la- will he in ortlor. .

la-I lm- :Itlt'mlnl to apologim- to tho who have lawn in-
t'nlln'niu'n- ml In lhi< 313: minntI- tit-lay. A tom-ting thn' rallml 
nf tln- for l? a.m. nlul l'vlt romla-lh-cl to
with-quip. I ?i m- Int-mayo thing-4 a littlt' rt-rzponsihly
at rh-l ll' iv. a - l-u ('uI'IL'tn- 

'l [it unm- --'Imla\ in ll-u- rli-tinuni :Ialrir-Ht' to tho
l'n- 

01' DR. 1.1212 A. SCIENCE ADVISOR TO THE
AND DIRECTOR. OFFICE OF SCIENCE AND TECH-
NOLOGY: BY DR. EDWARD J. BURGER, JR.,
ASSISTANT

lH'. In Mr. ('hairman, 1 Intro aHlu-cl my aanoriatv, llr.
llul'ut'l'. nl' in am who has llt't'll
tho to health and tho other utl't-ots of


\lr. t'hairmao. ham hol'oro this uolmniltt-o on April 15
on Ila- Fohjt-ut, and I am not; sum there in wry nan-h to
achl to what I .?filltl at. that-tinw, hot, thorn nro a low points I would
lilo' lo alnl t-mphaaizo.

Ila-n noon-thing ahont. the history and aml
rahu- of tho llH(' of this homicide and natal that rvrluw as: a Hat
from to draw what. to ho important gtvm'r-
alizaliona ahout. la'at irich-a. thee-'1' tll'l' of tho math-rs l. u'oohl
lilo- lo 

la'l. mo lit-gin by pointing out. that. ia a oholoival
Which has been introtlm?tal intontiolmll?,r into man's sorrountlioga
hocansc of the bene?ts to follow. Tta was to Ht'l'VtE
as an adjunct to other moans of wood hrnah contn? - I luml aml

(57)

nu ll -3
am.? :pgqm snmuumdxa go spupi all'.) ulsu Klulh?y A'uLu
J-Hls qs-Ipoul uuoq onuq saueddu sun allqm 
I 1011 04 pm:
AIM-11M) Hulnlunglxa) 3.11m; 04 q: puu
Km?; muq BILL J0 5:11:11qu
u: Mp?! Mun] mom am u: 1m? [unson?n?us'l

'suug-Jsanl) 
mum.- nsgu_.: paspu snug-maul: an? u;
?Imam: Am: *pmmum m: 'sown-m mum} .10} HllUb?I-idl muus
Amll pm: ulagnl.? 15cm: 1: pawns A'lnus HULL
1 mm: an? Maj-um: unguau'ju MN.) 01 ?uff}:qu
.IUJ mlgunusauh an? 10 unuo-Ismu mun? uir.) in?
?Aum ?u :11 ml u; unug-iuu.) [Llle u: 
I mung-pl man.? at? muss: an? Tm? akagluq Kinny. 

'ultussud
umuml .uaq.? Hanna 
?unit: In mun? gully 
?ll .IHJ u! lm-n Al. lulu] all" 

lulumgu! ?nth-[Hm]
Iln .Isll .IHJ Jn lull: 
Ju Hdn?ll .mJ Minguausg?dJ ?uugumn: ul
'saiuml 1pm: pm: .nlul m,
DSII .Inj .ll? JUJ 
j? [mpumlsus 
ml: mm: Ju 
JUJ-Imalnll HUM mam? lmgqaq m? ?anq ul
'MaJuy?ng ?llmaung, nuamng um A'q 
HHJILL UHII BUEJJH 
?lmy?! pup!? A'Jlumaog pull :[oum 
nu? suns.? mm?. HEHINI an? :m mm: '41

lm.n:.::l:lu Ilgxu?: m? usup mum: an? Hum Ill annual
u! A'anlunuul h'll pm:
letg'l'f'. :?ml lhuuandlug suns.? 05.11? Kuu: nun: ?Jowq u]
'53:ng 1m] 15"! m?
y: out? an? n: .CJunLuulul an? 10 1:10 Ilsa.? Ht: noA' (u IHHJUILLI 
man? BUDIIJEUH 
Ju 1mm:qu only?4mm? JI) mum-mu] m? JU
mm Kc. mu.? JD mung
?mu? pugJod Mulls 1: u! pan-aadxu mam HJOMHIIV 
.MII Hy? anu'?HHMu! u; small:
pm: Mum: mum: A'lamg-ullm 1: H: 
?up: Ju
?ml .numaq um: 1mm: 35:? mun? ml n: 1mm?: mu: ulxu?)
an?, mum" lugumml ugxu?: ml; pm: ?l?ufl-?r,
ml] Ju HHIIHHIHJHIILI m? 0  HIIUEMIU J0
ll J0 ll 
.u_xu  ?ug?m paupuun.) In!?
pupad I: 1m? ?Llamas?: uaaq q] 
Ju mph-um .mmu u: ml] Hm:
Jn 3.:5 ml: u: pnugmux.) Hams: .unu ml: .10 am?
"sawmx nun: pm] .manu.) 
?35.41:qu um? [mugmqn alum: an? lumlx.) .m




maguoo sum osuo u: mp: oql 'A'Jqsupu: A'q [mu wow-"own
mu A'q luoq pong-451:! 0.10m sogpms 03mm, 'sogouom: 50 dd l:
u: uun?oq mo.? [mug-mun: JJ-g?r?g uo pasmluq sung;
0:419:31 quoqu . 0:10:00 .101?: align-311:) m?
u: :mm p51: [n.mlm?q m? nu.? Mum: 1:1

0mm mtg-mun: 311:1.? put: sunsau [mu 
pupodxouu 'Amu J0 4:15am: m? 11:: .CnguJuclulo; HAN-aqua
Inn: 05:0.Iaxu A'mu m? A'qadoll.?
umgluulnom 1: 1m: sump mm? 1m? sum ?My u! plus 1 ml.?

J0 0mm A'q pnuLusn?! and.? slugud 050:? 
um.? 4: .mm anon.? amigo? 
gm: 5! unuuln?oJ .mJ, unusual .um 1m? 1m: poulyul 1
pun Mm: u: 
ml 45:11:: .mJ mun-X5 3m? 5:3? mm] s.?qu
'41 HI: su?suuylsaul: 
suuptm u: .mmu 011:: NV
'um um:
[um tp?nnlasml 1mm mm: ml 111:.) am: pm: nu! 1.10.45?
:[mm mm 03pm: JO spupl 11:: pm: slawl JU spupl 
mm: A'uu: 11 'Huqucl 1m.) nun? uu (.mqm
Am: u: mm: :4qu u: quL mutual!? .mmu
Km: 11:: Km: slmuznq an.? ml; sump: :Iugnnudujug 
whim?! [my mung: u: qugnd Km: am? 
INI
A'ulu u: pm: 5pm? Ilu
H-MIIHIHIIHIXQL n] .uml rpm.? 
45:11.50 shun Mam 1m? lmsa??us I mum? .m 
Mull .uau 5n pm: 1mm
?ml-mop Ju m? 1mm" u] up?: ml 1m: Ainsulmg
Ju :15 unlnaanpplus law! :lri'yl spuum
Jill) ?Film 51:? nu? palum I

A'q .mj mud [mu .(lnns 53:? .10 
m? ssuamd unnuululamp an? .JU 1.11M maul
puq :m $101?.qu JO A?lnmu Human.?
pawn?: u: 1: Kim-:3 55:? 'pgus mm] ?1 5y
'umrh: 1: .(puqs 5N1 mm] porilmu.) 
pnsudxn u: uugnuouu 
[mu many?: ?muunn .nlmuul u; ?mould-ml m? Ju [uglungml ml;
pnux?usaAug a: 1m? 1: lthlasaMchl Alan-:5 Haggauum HILL
Mn]
'u ml u; ?mulls pullmlumn 1m? 10 .?'ilugxuj alum: m?
uu pannuuamuw [Hill uu :lnauaw 
m? .10} A'pnp: ml: HIIM ?uldlunxa HILL
um? nuns-.1: pm: u:
.muma? 1: 51:11 pm: mm mm?: 15ml
Jo manna :muml Opussud mp annmri pm: .mnmm? u:
unusoul: (u un?oq mull 411.130.: Emmann
I '[ugJonuusgtl-1 1m nalmpuadnp
mum.? Jo Numb-ml 1: mu 1m 11"? as pvoJd sull 1!
m: 1: mm? pm: 

89

(it)

i - 

on file?spui ot' the. nanutnt tot accom-
i . (t inoit. systematic hasis and Without. the spirit of a crisis
necessary to urge. them 011 

li?tirther, one might aslc whether or not it ini-rht he. desirahle to
support. a. fairly sophisticated level of for a. 
nuaiherofpesticidal i 0

Now. I and others have. outlined the. research work on 24 
and 'l have. termed it. relatively sophisticated. Yet. I I
admit. that. there has heen 
nietaholie handling ot? this 

will have to
almost. no work done. to elucidate the

I erhicide in the animal organism. There
is little known Ill hioelieinieal terms of the mechanism of its actionti
and there is essentially no knowledge of anv possible 
heIWei-p this and other materials. I i
. in fashion We are poorly inl'orined ahout. tlto elniraeter-
is! as at the dose-tespouso relationshi i for very low dose levels. This
ot course. is the situation which we lace. in real life in the case of la,
ot' agentsminchidiug pesticide residues Item
the prohleia is a statistical one. In order to derive 
answers With any useful level of con?dence very lare'e colonies off-lex-
pernni-ntal annuals must he tested, often over a. lOII?rllul'iUtl oi? time

suggest. these voniuients to illustrate that. there are various
levels of sophistieation in research. I I

In the realm of pesticides the level of our
not. have kept. tip with the state
level til. questioning to whit
seeking answers.

the very exeelleiit. report on research needs compiled hv an advi-
sory task force to the National institute of l".uvirounienial llealth
?ea-tires outlined these research areas verv Well and WW explicitlv
lhis report. i not iul'oriued. is just now puhlished.? I I

.\ll of this-"more sophisticated research.
reseat'i'h slitiltstit'eil the 
repeat. it' :i really serious thrust is take
oliligateil to that new institutional av
research since. as a

research activities may
at that. art. one With a corresponding
-h polier makers and the public are how

ioore expensive. research.
ill cost. money. .-\gaili. I
in this direction we. nan-lie
I enues for 
part at the rust of development, the hill to
may he higher than We might. desire.

_of this discussion lirings me once. to a point. which I
made in_in_v [it't't'tiills testimony and whieh I feel is worth euiphasi'l-
inLr. ll htle We as :t society ha ve reventlv heguu to ask more penetrat-
ing questions aliont the [ins-:ilile adverse health etl'eets of environ-
na-utal agents it is not clear that We know how peiielt'aliti" this
questioning should he or must. he. I

What. said la-t'ore was that. we had set. our sights hitrher \Vh'it I
should add is that we. are not sure. how high Ithev should he'set
For example. tip to the present. time we. have. lieen williu-r to live
a the amount Hf 

perto'rmed on a. pesticide was to some. extent. related to the prohahil-
tt_v ot human exposure. ith a low or negligihle prohahil-

ity of exposure. relativer little. understanding was sought. mid little
research was undertaken. in fact. one could argue that since. the
appearance of residues of have. heen rare. events?it. is very
rare to tind measurable residues of on 't'oodmtherehn'e one

til I 
did not. have to linow too nuile ahout the. toxicology of the heil-I- lag

- ve are more. tartieulat'. I I I
 til we shoulil he more explimt alioiit tissuiuptioats it?s: :2
these. If they are valid they stand on their own men "um My
are not. valid we. should change them. I am hapr to Enigma.
otl'iee. is examining questions such as these at. the picsen . 

Thank you, Senaltor IHart.

Lt: am: on.

 suggest. the possyihility that the lierthl:
assume a. greater role in any testing area. but. speci ica 
matter of pest icidqs.

- es. . I

llilhlirr. You indicate that. otherwise the hill to 
might. he higher than We. might. desire, which 1' suppose ip w"
wav of saying an industry could not. atl'ord it. on sio

0 I 
ml?JIrilIiidhuotiic. \Vhat I meant to say is it' industry is retpuret: 
carrv on vears of very an)r pew 
can he ohvionsly industry Will no longer loin tilts?im
in -niamit'aoturing new products he "aitse. they could Ilit'tll. lltutrul 
loss. {has the community! the society a-vould he. ro I'll'tl lot 
future. verv valuahle chemicals which society'uuglit him lis'ihiimn?
health and other reasons. to impose I. a; 
a particular company that- hetero it. ran a pint Iltr.  
undertake millions and millions of dollars more uslta 
than it. has in the past. Would simply stop the development. in . ilieit
products. 'J?hei-efore it. seems to me. only fair, sincewe aunt, to pin it.
sot-ietv as a whole. hut. also to encourage henetits to 
sm'ietv as a Whole. ought to particil?mte in the eost..of determining
a hat 'Ihe damages may he as well as what. the henehts may he.

Sea-nor ?l'hat. then would he your haste answer to the.
Hist" ti-m Hr argument that research is just. another element. of the
3 hi -l3i-'liI-ti.

ltt'. l?t . Yet,

ll I. 'll '11 tlH? Il?i't' ol' the product hear that cost.


?Flinn-hi. I am not that. the. manufac-
turer should tint :Il-u lu-ar  tautial t-o-ic, Ite?shuuld sure
that the product that he pimp-n ill:? to not dangerously
lI=I\t' ulntmi- udu-r-?e human health etlvt'lf?.
ltnhi Irv :hoilhl he required to undertake a reasonahle set. of expel-i-
ineiit-'. and lhi-v do. to as-ure. that this prodnt-I 
their mind Stilt-Iv factor. llut it may take ."ears to tind out low
level. and eiisilv hidden dangers ?'llit?ll suineliliies lit-comp ?twp
ous onlv when mass use. is underlain-n. To help nvtiitl these dangers
l'l'inkl Wm" ill research program Would
he. tlesil'ahle.

Senator Do von lmow whether the. administration intends
to einharlt on hroad new research programs in this area?

Dr. There. are several agencies which up" (towinl?ng
plans. pursuing resent-eh in this area. and helieve. an 
committee. of the Department. of NEW headed by Elm] 

sacramenea .. .. 

0'2
is making some proposals to NEW ahout substantially extending
its research and testing activities in this ?eld of pesticides. and
about. developing extensive facilities, large animal colonies and large-
Seillo testing equipment. and peisounel, to carry out extended tests in
this tield.

These are that are being formulated and are
being proposed to HEW. I do not. know what the status of them is
at. the moment.

Senator .llau'r. It is likely you would not have the ?gure with you,
hut. let me ask the question. Perhaps it could be provided for the
record and have no idea what it Will show. lint would it he possible
to identify those activities which are not undertaken by the Depart.-
of Defense that are. comparable to the. line item researeh and
dvelopnient that is done. liy the Department of Definme? The total
tigure for research by the Department of Defense is in the range of
$7 liillion a. year. How much are we spending in other research?

llr. The total Federal budget for research and develop-
ment. in this current. yea is close to $17 liillion.

liieliidingthe 

l)r. liieluding the H). ll'yoii take. somewhat over
$7 liillioii of Hill) l'unds. it. is $9 liillioii to billion in all other
branches iil' the. 

Senator ll.\irr. We will let others judge whether the allocation of
the resources is or is. out prudent. I think it is good to have it in
the i?eeoril.

Yesterday we testimony from the Department. of Agri-
culture. When we think almui i-hanges in the system of pestii'idc
l'i-seareli and Madrid we have to think :ihoiil rhaiiges the. hasie. law
of pestii'iile. regulations. the te-liinonv yesterday from
the ol' .-\grieulture were a another of suggestions and
changes that they he. made. in the. horde :ii'l. yon
with these suggestions?

Hr. Di: llr. lhirger told me ahout. them this morning and
have a. copy of them here, and we have. discussed them. 'l'hese are
in line with some. of the. suggestions we have had with Agriculture
and other namely, that. there is not. llexihility in
the present statute to late. a action in all eases where new
in l'orniat ion heroines availalile.

Sometimes new like the study is very sug-
gestive hut. he. said to he linally eonrlusive ol' the
small minth of animals and small of (-ireiinistauces in-
rolved and the separation of an impurity may hot. he taken care of.

It will frequently happen that. _\'iil1 will get. preliminary sug?
gestive results not. sullieient to aholish the use of a chemical hut
siitlieient to take some prudent action until more ?nal results have
lieell nlii nllit?tl.

I think the Agriculture suggestions do more in this direction to
give. for example, what they eall preliminary suspension authority.
So if a danger sign is raised you take prudent. temporary action and
continue further research. If the further research con?rms that
safety is there after all you can remove the suspension. If further

resea' proves the danger, then permanent. suspension can be-



i



CD
31?)
ill 
. CD

Senator did not. quarrel yesterday with the Hey-ailment 
Agriculture's interpretation of the hash: art. I lltl\" lie d.
as we always do, to justify what we did, namely, those of. iiw' who
wrote the law. 1 am not. convinced that that the
Department of. Agriculture, when, as you say, the Signal goes up
from suspending the marketing of the product?I am not. sure that.
Congress should he held to have said that we excluded the possibility
of a temporary suspension. 

Just as I was not yesterday, I am not. sure you are not equipped,
and did not plan to dehate what. limits there are under the esist 
law. .

Dr. lHilhniinu. No, I am not. an expert. on the question of
interpretation of the law, but it. has simply lieeu taken for granted.
niayhe not properly and maybe under an interpretation that. should
he further developed, hut it has been taken l'or granted that pre-
liminary or temporary suspension was not provided for, explicitly at
least, in the statute.

If the law were interpreted to allow this, it. would he line.

Senator llAlt'l?. I cannot imagine there would he any dill'erent
criteria for a suspension than a suspension. 1 still have
the feeling that. the suspension is not for a. thousand years. If you
suspend and then diseover that your alarms were groundless, surely
you can uiisuspend. That would argue that you can temporarily
suspend.

Dr. it that. is the ease think that. is line. it. apparently
needs to he. made more. explieit to the. people who are. doing the sus-
peusiou they do not. feel that they have this authority.

Senator llan'i'. (?learly they do not.

A re. you yet. in a position to advise whether you would support or
support of the suggestions for law changes made yester-
'l Agriculture?

I think it is fair to say that. we would lielieve
ll are proper mores. Again, as I say, we are not experts in
ll" Iii-Id and the legal field. ?that we tried to do is to ?nd
I  'ilel lea-linology that is applicahle, and we leave it to
the legal authorities to determine what. rpm-ill.-
unluinqp

hot I ilii: additional th-sihilitv does in pi'iin'iple sound \erv
desirahh- to no, {euph- results, you know. are in-ier
the. final answer uiile Ila-v are t'Vll'l'tllt'lV eonrliisive. results of
CNlI?Otnt?. danger. I 

it is alinost never poi'sihle Io sav that all the i'esi-arrli has heeii
done and it. prores that. the, thing 

?enator IlAirr. Conversely, forever te.

_l here were several other suggestions that Agrienlture made. Hue.
regarded the ineffectiveness of handling push-M? In.
'.lhe_v.were going to recommend rest run-luring the law 
require a grading liy degree ol? the hazards of a pesticide and to
ensure that products would he permitted to he
handled only by indivnliials or institutions licensed to do so. Do vnu
have any comment on that one? . 

Dr- DUllmnuu: I tliinlc_I have no very expert. or tent. exit-pt.
to note the experience Within my own family that lahcls on packages

I

unn



are often not. inleipiately read and that. the laheliag prohlein is a
dillirnltoiie.

Henaloi' llairr. As Agriculture indirated, not a very relialile handle
for protection against. injury to health or enviromaeut.

Well, we would hope that. you will lend your distinguished scien-
tilie support. to the ria-omnieiulatioas that. Agriculture has made.

A couple of other questions, aiul this goes to the ditl?erent actions
that: have been taken with respect; to pesticides or herhieides by dif-
ferent; depart meats of the Government.

Agriculture suspended I think you said ahout ?20 percent of the
use. or 

llr. Dottiania-t. I did not intend to imply that was all .'\gricnlture.
I said the total suspension amounted to ill) percent. of the total use.

Senator Ham: ?2 is my understanding that the Department of
Defense. has suspended it entirely for use in Vietnam.

lh'. lhilliunia-i. That. is earl-eat.

Senator And the llepartiaenl. of interior very 
suspended it. for use on pnhlie lands: lands in its ownership.

ttr. llutliama-i. Yes.

Senator How do we. explain the ditl'ereat reantioas from the
several departments with respert to the same. product?

lti'. I do not know that i ran fully explain it, hat. the
situation in military operations is not m-ressarily the situation in
normal peaeet'nl operations ill this ry.

lii military opt-rat ions. liy iieressily. these elieniieals are dislriliuted
liy and it: is not. always easy to eontrol where they go and
how faraway they ldow or even that the airs-raft is hitting the. right.
target. 'l?heret'ore. i think somewhat more priidenee may he required.

?'lien you are doing it. in a. normal way. in agrii-nltaral praatiee
or land management, you can he nan-h inore raret'nl and make sure
that the material does iiol. get. on food erops or in your waters, which
will rontaminate the waters, or get. on grazing lands on which aili-
nials will he grazing amt therefore. get. into milk or meat.

ti'ortaiiately degrades quite rapidly. It. is not. like BUT.
it you spray a pasture with all lraees of it. are
gone after ahoat three inouths? exposure. to wind. rain, and sun.
'l'hei'el'oi'e rattle. quite safely graze. on grazing lalid treated
with a ti er this period. .

Also, if- if, gets mi il?. is likely to degrade pretty rapidly,
though it- is desii'alile to have no tolerant-e. for food. litll where it. is
used in areas where there is no human population and where. there
is no danger of food or Water. then a eonlrolled use
run he. extremely vatnalite. and since. no dangers would he resulting
i think it. is perfectly proper to have. eontrolled use in this country.

Senator ll ??hat. do you say, then, to Interioi"s prohihilion for
use on its lands? 

Dr. linthanoa. I guess I am not familiar with how extensive that
use is.

Senator ITan'r.
yesterday. . .

Dr. Delinuma. see. I am not. familiar with the background for
that aetion or the extent of it; or mayhe they found that. other me-

I am told that Interior?s action was taken only

.- 

., 





to .

436v

terials roiild he used on piihlie. lands other than 2,1 wlneh would
serve the purpose. i just. donol know. - I I

Senator Ital. it. rontrihutes to the uneasiness of the polite:
when we see these seemingly ronllit'ting reaetions. 

Dr. Dultiiniei-z. This is one. of the very delicate things that. the
Government faces; that. is, not. to overreaet to Situations which Will
do damage hy ove.i'reaet.uni._ lint. there is an opposite of
under reaction and not. taking prompt. or admaate action when
dangers are evident. I would like to defer to tie Department. of
the interior in this ease, aml Willi your permission, have that. He.-
pa rtmeiil. sahmit. a. statement for the. record.

(The informal ion followsz)

oN Pasta-u?: t?oucv

TI..- ln-parinn-nt ot' the liitei-liir pulley stalenn-at, Issued by Heerelary Illa-kel

on June IH, does not itlt't'er markedly from the one that has lu-en l'nltom-d for
several rears. Nevertheless, It. adds some elieaileals ahnal wtih'li We have i'erelved
inlilltloiial itata the last. year aial ri-enigntm-s tlie eomern expressed by
-- h-i tin-at. oi 
lltliu-tll ot? the hydi'In-arlinn tiiseelleldes had
been in elTeet. for several years and that; no star-e tietola-r Juan, 'ttie
hazards til' lit?t'" N't'nutilm-il for some time were 
hv amnltortna results In the last. or eight months. .\ia trol was tin-tinted he-
film?! or tile ot? the l-?ood and lirng to the use of
eai-elin-aeos. 

'l'lie lit-wartinent of the Interior pulley Is not Intended as a Federal pestlr-idu
poliv)?. ll Ilia-s loin: lil?i?ll the pntiey ot? Iii-partnn-at to set a standard in its
use of pi-slh'hles that. is heyoad from the standpoint of safety. (,nnse-
our posltlon may he more slrh-l than t-?tltlit' "Ila-rs ulsh to set. If
demonstrates that snow- of the Il-ti-il may he hm: hazardous
than \ve suspeet; then they may he reslori-d l'oi' tr on lltlI-I'lut' lalnls.

Heaator You reminded me of :i (pie-Jinn kielaallinyself
yesterday for not. having asked the lh-parlna-nt. of .\gi?teullure
Wit Perhaps you ran help. 

You un-ntioneit- lili't'. I rerun the testimony yesterday,
under the it?llt?lt.\ a sc-irnre advisory is eitlaldisheul
when there is a eanei-llation proredul'e aimed at. a pi-oihn-l, 'l'tn-y
explained that. in the of DDT that. some six inonlhs have passed
sinee the ram-ellatioo proeeduro was and no eoininittees
have here l'ornn-Il: hein'e. the iassage of time has heen extended at.
ltl'sl- lii' this amount during whieh. under the. prim-?lures,
marketing oli the product. goes on. ??hy the. de-
lay?

lti'. lti explain that. do not. know why there
should he a long lag lielween these two events. The only thing i ran
think of is siuee. the. Heerelai'y?s peslieide. advisory eoniniission has
lieen eontiunoasly at. work on the peslieide. prolih-in. espeeintlv will.
attention to they were depending on it. to evaiuine this par-
tieular prohteiu.

Senator Ilan?r. repent, I should have asked them yesterday 
(lid not. lint. if they read so strictly the. statute with respeet. to
suspension. I would assume the same st riot. reading would tell them
that. they snlistitule the Mralc Commission for t.l- explicit
statutory requirement. that. there he a science hoard estal .ed for
each of these products.

Mnn_





 (36

llr. Du tuition. I would agree. I think maybe the law does not set
time at which the science advisory commission shall be established.
senator I-lMi'r. No; it does not. It was assumed that it would be

ahiislied at least with all delilicrate speed, and 6 months seems to
delay.

Eli'. Duilinnuii. Yes, I agree. \Vith your permission, I would like
ask the Department of Agriculture to provide an explanation of
apparent delay.

information followsz)

Daranninnr or Aomcomunn,
Ol-?l-?tCI-i or run Hi-ieasrAiir,

?ti?aiittington, July 1970.
A. -
Secretary. Council on Environmental Quality, Executive Oince of
. the President, ll'oshinyton, 
.l-Mlt. int. in reference to the question raised at the "art
imiltce hearing as to why it. has taken so long to establish nu advisory
uiitli-o on we submit: the following explanation:

i) After the of the companies involved did
request an advisory committee or puhlii: hearing for the four uses of DDT
re cancelled until the latter part. of the till-day period provided for appeals
lltl' Federal insecticide, iiliil ltmlenlh-lde Act.

?ii Meetings and discussions were held with representatives at the National
dein,v of Sciences on makeup of the committee.

Previously we had asked for lists or names or persons to serve on ad~
r: committees for other cancelled products, and they were already in the
'05s of compiling them.

it line to uni'avorahic piihllelty related to ciiniliet-nr-interest charges re.
tini: authorities that served as consultants to the Department in the past,
experts wri- not. willing to serve.
--) 'l'wo additional requests to NAS for candidates to serve, heshles the
had. were necessary in order to complete the 

Two of three companies requesting advisory committees withdrew,
in: only one for a tracking powder use.

i) (letting written position from Dill-2W as to whether tracking powder
considered an essential use from puhlic health standpoint.

it and getting approval of proposed candidates to serve on the


Notifying committee members that they were selected to serve on the

t?outllet-nf-lulerest review and evaluation within the USDA.

Sincerely,
tin-Lair,
Director, Heir-ms! and Education.

euator Ilau'r. would hope. in connection with the cancellation
_--eediug on an advisory committee will he. formed with

his next (nestioii hears on your liroad liackgroiind. One
he ditl'ieu tics in this story was the diil'iculty experienced
information which might have. heen of public health sig-
-ani-c.
fiider what conditions do you suggest scientific information ought
it? kept secret when a question is raised as .to the safety of the
ltti'l 
Mir. Diillinncii. Tf you are talking about. scientific information,
i not. think it should he kept secret. \Vheii you are talking about
having to do with the manufacture of commercial prod-
. that is a very ditl'ercnt situation because the costs of develop-
t, testing and ?ing a commercial product into production are

hit:

. r-

u-rv high. it is quite proper that the litilltliidl'lIIl-u .-  I
In! Ilil'lilt?t'lt'll so that. he will liaie. old-rottinin too-voter 
curls. should have. a [silt-til. or it protection loi' o-I'
his product so that he can recover the very'lai'ge costs to den-lop
it. Whereas, if he instantly pulilisheil all the. information shout how
to make this product so that. others would instantly st art. making it.
without. the expense of' development, this would ohvmusly he an
unfair kind of competition. 

t)ur whole svstem is based upon the fact that inventors of new
processes amt products have protection to regain their investment and
recover their costs. 'i?herei?ore think the puhlication and the dis-
tribution of information with regard to the manufacture of products
and materials is a proper trade secret. 

()ii the other hand, when it is clear that human health is at stake
I would assume there should he mechanisms by which Government
agencies in proper authority could he told something aliout the. coin-
position of the product so that. they woulddio ahle to determine
whether or not there might. he the product which
ought to be investigated for their harm. . 

I do not know exactly what. the law is on this, but it would seem
sensible. 

Senator I-lan'r. Your suggestion is that when a anestion of health
is raised with respect to a. product, data and information on the
product should he made availahle to the appropriate 
agency for its determination as to a question of health and safety?

llr. Yes, sir. 

Senator That excludes, of necessity, the judgment. and the
comment of perhaps very gifted men aml women of science in or
?ring at the determination of whether public health is or is not. in
This is not to suggest that the appropriate agency lacks
and competent people but surely they do not have a ino-
l"'lmlr on that. I

there some way, notwithstanding the obligation to protect trade
Is aiul encourage invention and discovery, we can do a hetter
joh of permitting the outsider, whether it. is the. head of llio_chein-
i' 1 [Hit'ltitPIIl at. (?al 'l?eeh or someplace else, heing in and
an opportunity to sharpen the. ol' 

l?l'limialzi?. i. think that the various advisory mechanisms
to the various (iorernineiit. agencies ought in general to
that A science. advisory group can he. called in
to consult on it particular pi'olah-ni. on the. possible. dangers of the
chemicals that happen to he. in a particular commercial
product.

I am sure that. they could tap the rest of the scientific community
to find out whether A or chemical is of a nature that it,
would likely he harmful. 'l'la-y do not have to reveal the whole con-
position of the product in order to say this product happens to con-
tain a certain amount. of eoiupoiind A, is there any evidence or
am; chance or any reason to helinve that. this compound A is harm.
fiil. think the knowledge of the scientific community could be ob-
tained.

Senator lian'r. Should he olitained?

Dr ?ailerons. And should be, of course.

(?593






Senator Ilsa-r, The suggestion has been made that all of us as a
pen do would lie hotter ntl' if we had some centralized clearinghouse
or data hank into which could he fed all of the information not. suh-

jeet. to trade secrets. ?are you given any thought. to that. suggest ion?

Hr. ltultnnuin. underst and that. ltr. h?leint'cld is going to dint:th
that. question when he. appears.

Senator Your reaction would iiot. stop him.

Dr. He knows so much more. uhoiit it. I do not want
to anticipate it. It. is this question of the dissemination of scienti?c
information which is one of our most. ditl'icult'. prohleuis. .

In past. years we. always assumed that. if you published in scien-
tilie journals and hooks that. anybody had access to, anyone would
just. go to the lilirarv and look up what he Wanted to know. The
volume, of seientitic information has liecome great, the. urgency of
timling nl' information quickly has heeomu great,
and this has led to the question as to whether one. can or cannot;
use modern scientilic equipment. to store. and retrieve scieiitilie 
formation more. expeditiously.

The only troiihle. is it. is very expensive, and the development. of
techniques for putting it. in suitable form for data. processing. the
question of how agencies should lie. involved tn feeding the
information in and mw to get. it out these are tech-
linlngieal wliieh together with the expense have not; been.
Worked out. i think it is a Very urgent pi?ohh-m.

There are. people in our ntl'iee that are working on this and I
hope. we ran timl ways to have a storage of needed infer-
particularl_\' in the. health tiehl.

Senator I share. in that. feeling.

My last. question. again, is a general one. had an exchange
yesterday the thipartment. of Agriculture witnesses on the old
that. is ereateil in the. minds of some. when you have a de-
part meat. that. is charged with the promotion of an aetii'ity under-
taking also to regulate lii this case. expanding agricultural pro-
duetinii is a respoiisiliility of the. department, and iit. the same
time we say make. sure that. herliieides are not. permitted that. do
damage. lto you any general rule as to whether promotional
and regulatory should lie separated?-

Iti'ltiziinaz. I do not. think I would propose a general rule on
that. In the partieidar ease ot' peslieides it. is true that. is
Illtill't' lltl' itili'l'ngt'llt'y :Igt'l'i'lin?ltl. 
and \i'ell'ai'e and Interior aet jointly with Agriculture nu these I lies-
tious. ltthere is an agrieiilliiral produet that is in use and has
that. there may llt' health etl'eets from this, they can
immediately it. to the. attention of and action can
he taken then hy joint. agreement these three departments.

I think this is cute. purpose of this interdepartmental ar-
rangement. I am sure. that: no Department. is anxious tn promote

something that. has danger tn human health, and as soon as human
health aspeets are nut. has the. ohligatiou the au-
thority to this to the attention of any other Department. In
the ease of pesticides they meet. and take act inn jointly.





i ?9
-tn- \ii'r. .nnun fellow Votes, and that Ix tlu- it. Li H. ea:
?mun? l' I'I-riw-t. third. it?.ml
Hr. I?m, 1 do not. thin in t. . "n
has heeii agreed that. d?
[or speci?c action. 1 think this .is piilia t. .
notdniow whether it; has been made o; itililivi?u wow 

Senator Ilaa'r. It is my. understate mg I . . 
the decision would lie Agricultine s.

 You have told us that. the Liepai-talieiit.I ol' dig; we?ll-l

tum docs ?Ob believe limb t'Ihclv and
like to see tiem iai'lave they to you, as tlu'iry till: 
yesterday, that. iielver ?mi. .Hm
?t nus insticn (ill (inol' lt'ood which had lieen treated by that. ?as
- sto I
dilii?'ilighltinooa. When there is clear cwdence of 
they have. full authority, of course, to nr 
The prohlein enmes when the is not'l?liat'. was my impression as 'well. tut. 
that the use. of: a pesticide on food crops will 1.13mith
hazard to the. public hecause it. take   
for that: food to arrive on the. tahles of those peep to i 
and therefore that. the. hazard created is not. in l-tlt'l. .1. m1-

Di'. Dulhunci-i. 'Well, there is a legal_ determination am .111 It ti] 
tation of the word ?imminent.? Sometimes food reaches .l ie

we . sometimes itdoes not. I
Can we impute to Congress the intent. toI 
iltlillt'. unprotected in such a. case? As aunt 
legislative history on the use. of the. 
ai'ul" in the relevant?. net, and Congress has not. detuied t. in. ltltit

in? '1 
As I understand it, hazard leads to one
kind of m-Iiun hazard present. hut. iiot'. imminent. leads to a dill-
t'l?i'II' of (hie. have. either suspension or raneellat ion
5? Hint-ensiun oeciirs when there is imminent hazard, and
it sounds more. temporary. really is not, I

lt is not. given the. eaneellatlnii prneudmcs
mav take as long as rears. 

'l'hei' take time. So. llit't?e run he the,
case of imminent. hazard. then- run he if there-is hazard
lnit'. not. I do not know how you interpret. 
whether it. is a ?luv. :1 Week or a mouth or what.

Mr. interpreted it. so that. they would not he allowed
to more in the ease. of known hazards to human health when the
hazards result. from the. use of a hazardous pesticide. on food crops.

Dr. DUBnmim. Then possibly a elarili :ation of the statute would he
desirable.

Mr. llu'icwi'r. I just. wonder whether it is Ilt?t?tlt't
fact. that it. seems perverse to to that; 
tend to exclude protective action in
Dr. I?lense
legal counsels for the
various eonrlasions
under the law.

Mr. Iiicicwi'r. I will not if you do not. ask me. to understand it.

I have just. one other question which relates to a legal term, lint. I
would lilm to hear your reaction to it from a seientilic standpoint
wit liout. legal context. attached to it. 

Do you hare an},r reasonable doubt. about the safe
Btlt'll as and Hilrex?

l)r. lhillauau-i. I have reason
the research and testing 
qeustioiis of substantial here not. heen

proved so I would he much more comfortable nho
than I would ahout where the teratogenic 
estalilislied. For these others, I think imminent.
have not. been found.

Jlr. IilL'Ith't'.

Senator Ilaur. Thank you.

Dr. Burger, is there anything you want; to add in light; of our ex-
change?

Dr; ?mama. No, I do not helievc so, Senator.
Senator IlAlt?l?. I renew my apologies as I ask Dr. Steinl?eld to
come up. I lcnow there are

many things he would hope to he. aide to
do this which have llt?t'll delayed.

Dr. Jesse Htuia l'eld, the Surgeon (lain-rill.

in of the

loiigress would 
rases such as that}.

do Not?- ask me. to understand the views of
various departments, or how they come to the
as to what. their depart nieuts can or cannot do

ty of pesticides

alile doiilit. about anything in which
not. been adequately carried out. The
dangers in those cases I ll
at the use of those
?eet. is now clearly
or serious hazards

Thank you very much.

STATEMENT OF DR. JESSE SURGE

PARTMENT OF HEALTH. EDUCATION. AND ACCOM-

PANIED BY DR. PAUL KOTIN. DIRECTOR, NATIONAL INSTITUTE
OF ENVIRONMENT HEALTH AND DR. WILLIAM M.
UPIIOLT, ACTING DIHIZUTUII,

PESTICIDE
ADVISORY COMMITTEE

ON GENERAL, DE-

lli'. Thank you. Hi-uati-r 
With me this are Dr. l'niil who is the llii'rrtor 0f
tho National Institute ol' Hi-ieiu-rs. He. is on
my left. And on my right. is William Ii'pholt, who is the execu-
tii'i- secretary of tho Secretary?s Pesticide. Advisory 
lit?IIll'O I lit-gin, I would like to apologize for a number of 
spelled words, ruu-ou sentences and so forth in the statement. of which
you ham a copy. It; will lie somewhat. dill'ereut. than it; is before
you as I read it. if I may be permitted to read it.

Senator You may he almost certain I won?t. spat. the misspell-
mus.

Dr. I am pleased to appear before you today to discuss
the actions that. have been taken to protect: the public health by
the Department: of Health, education, and \Velfnre regarding the
herbicides, particularly and EBA-slietleiul .. . ?ql liv

. ?(In to ?mm, Ill .
of hazards in l-llt- Wul hm.? ?mite,
and lil'm- ?ml?
Leven in the extremely short: tuno bill -a
.riigjl'Ulibi the int-ch
"I?W'umyiihiv how hest. to uudeilal?c hazards

t" .1 ?q?mll? 

. - . -I lul'y . 1 hiqn'ulnu. :tilt

-- "All" in l.ll)tllc  - '1 - It is certain .
lm?dn?t i .1 instienrh. - - i ?nu theestral?" r-
posed lIlul'ty  animals very lugl'itltl to Very
so 33' Him" in which man .13 exposm In. 1 ?ma. mnlamsizc that.
ol? these chemicals. I?llui-ies of mammals. 
. l'H-q Wt) - . 

- -. lilih?utl bu?  -. - ies when 
'wll'lh  as to tho of those stud
certain - . Plug.? llilii?inl'oriuation on the, lirall-ll Mimi's

u? agencies Wit ?1 Ii: 1\ ?my.
lic 190410111" hum? l' niins'tiididrs is 
In"  fill: ml  and the Food nqu

1.1 Ji ii?nl't' i . mum,? 
- . l'li tliel?mtioi41?" . l~ to
tilthis th .
I .rir-al information and ate hiuldii ml ?'13
Sllm'm" tin? "h . . ltI-is'ittl Willi l-lit?. 

Ioriu the clearinghouse. I cm Wt)!  
. ilel' to dnlt?. . . . llmt?
Drug: Adanaistratioii has issnelilt?iliisiq gm?u

t' ii lo the extent; of availahle I'iir tlilB
special atteii .io I. ?i 2.153]. ?n rum] ?mph I I
ill ?It! [Ul' H?hlt Ill-I I ?l fur rl\hiH ?ins :th 
'ide was Wiimm? I hi insure. to residues til?
m' to ii'event, iireldrli a1 . .. 
additional precauth .utl'H-st ind not. deteiliI - 
even tili?llL'l' ("Ir Sim? I?t?i-T on llwsu foot CI "lilt'tll was
ins u, a the "mmumi

'l'la- seieutil'ie research on -

Ire-ml liaseoiitilincdmentn t-l . .

'lil'? Nl?l'm'nl In?tmlm ?f related 
rl'l'u'm? ml Mil",- . \iu is the dinxm
"117 
we In in his-I of ?t and 

Additional ?in. ll 'lrt: the earlier 
dioxin in the l'rml- "will llu- minim. lliie Milli." 
t'lt?ll I'll "ll'lmwul'me'm and NW Ll.l.:i-
Utilized a pnm?mmlm? ?fl lii i-tl'i-i-l. of these?Hill'le ?ml I palate nioiiseinitiated with esters or [a

studies "namely, the li? lqgun iq the. same. as that ii:-id tn
eol-laitvl-ester. 'l?lio 'll?lull-V fun" or to date. are. suggestive that. at. least.

The results that 11"" 1 1? in tor-Ltniretiic actirity to that. of
. . be compnla in . I In?
Him? ?htch 'y l~linitive studies on these letlh 
At; this time mul?c ?9 
underway.


I I litilllt'IUlt'. 


  

- reason?
.an Associa-
a project?
3 ways the
auxin mole-
-.1bout how-

about the
Iprecursor]
:n or wood
red by the
[5413? when

 
  
 
  
 
  
 
 
  
 
  
  
 
  
 
 
 
 
  
  
 
 
  
 

33*. regards
anybody
'uiicated to
to an out-
it would
nt dis-
tests show
Iii/kg and
.1 the chick
ddent," be
all 2,545.11

however.
74 National
glefects for
9'8 wrong.
limits dole-
*1 of mal-

  
 
 
 
  
 
 
  
 
  
 
  
  
 
 
 
  
 
 
  
  
  
  
 
  
  
   

50 Studies
defects in
?Ge ama?

JEICII: data
helps,
?1 DeveIOp-
on

lune 8?11
"Id 20 gal-
ert'
goctors
uck
?Wt: 9.3
and one
. ?f Tonto
?9 'CODter
:4 convul-
""hosltosis,

hell's who
Do?toning

vouch.
t?Se. that

?lls pain
"g ducks.
. l1ins of
?1 as
any of

 11 cases
N15 and

 

 
 
 
 
 
  

- -I.-I nun-uh.- ?Hmudh'mm?l

 

107

I'm no toxicologist. People here are emotional and each morning wake up with

 
  

 
 
 

new nails pounded into their palms. What's needed in solid scienti?c investiga- 

tion. All I hope is they don't leave us hanging in the air for the next 20
years." a . .

Appendix 3
[From the New Yorker, Feb. 7. 1970]
A Heron-res a-r Macs: DEFOLLATION
;By Thomas Whiteside

Late in 1961. the United States Military Advisory Group in Vietnam began,
as a minor test operation, the defoliation, by- aerial spraying, of trees along
the sides of roads and canals east of Saigon. The purpose of the operation was
to increase visibility and thus safeguard against ambushes of allied troops and
make more vulnerable any Vietcong who might be concealed under cover of
the dense foliage. The number of acres sprayed does not appear to have been
publicly recorded, but the test was adjudged a success militarily. In January.
1962 following a formal announcement by South Vietnamese and American
of?cials that a program of such spraying was to be put into effect, and that it
was intended ?to improve the country's economy by permitting freer communi-
cation as well as to facilitate the Vietnamese Army's task of keeping these
avenues free of Vietcong harassments,? military defoliation operations really
got under way. According to an article that month in the New' York Times, ?a
high South Vietnamese of?cial" announced that a seventy-mile stretch of road
between Saigon and the coast was sprayed ?to remove foliage hiding Commu-
nist guerrillas." The South Vietnamese spokesman also announced that defol-
iant chemicals would be sprayed on Vietcong plantations of manioc and sweet
potatoes in the program was gathering momentum. It was
doing so in spite of certain private misgivings among American of?cials, partic-
ularly in the State Department, who feared, ?rst, that the operations might?
open the United States to charges of engaging in chemical and biological war-
fare, and ,second, that they were not all that militarily effective. Roger Hila-
man, now a professor of government at Columbia University, and then Direc-
tor of Intelligence and Research for the State Department, reported, after a
trip to Vietnam, that defoliation operations ?had political disadvantages" and.
furthermore, that they were of questionable military value, particularly in
accomplishing their supposed purpose of reducing cover for ambushes. Hilsman
later recalled in his book, ?To Move a Nation," his visit to Vietnam, in March,
1962: had down down a stretch of road that had been used for a test and
found that the results were not very impressive. . . . Later, the senior Austra-
lian military representative in Saigon, Colonel Serong, also pointed out that
defoliation actually aided the ambushers?if the vegetation was close to the
road those who were ambushed could take cover quickly; when it was
removed the guerrillas had a better ?eld of According to Hilsman, ?The
National Security Council spent tense sessions debating the matter."

Nonetheless, the Joint Chiefs of Staff and their Chairman. Generaixhiaxwell
Taylor, agreed that chemical defoliation was a useful military weapon. In
1962, the American military "treated" 4,940 acres of the Vietnamese country-
side with herbicides. In 1963, the area sprayed increased ?ve-fold to a total of
24,700 acres. In 1964, the defoliated area was more than tripled. In?1965, the
196-1 ?gure was doubled, increasing to 155,610 acres. In 1966, the sprayed area
was again increased ?vefold, to 741,247 acres, and in 1967 it was doubled once
again over therp?revions year, to 1,486,446 acres. Thus, the areas d'e?foli'a'ted in
Vietnam had-increased approximately three hundredfold in ?ve years, but now
adverse opinion among scientists and other people who were concerned about
the effects of defoliation on the Vietnamese ecology at last began to have a
braking effect on the program. In 1968. 1,267,110 acres Were spra and in
1969 perhaps a million acres. Since 1962, the defoliation operatic have cow
cred almost ?ve million acres. an area equivalent to about twelve per cent of
the entire territory of South Vietnam, and about the size of the state of Mae'-
sachusetts. Between 1962 and 1967, the deliberate destruction of plots of ties.
maniac, beans. and other foodstuffs through herbicidal spraying?the word
?deliberate? is uSed here to exclude the many reported instances of accidental

45?362?70?8

 4

CD

..

 

  

. an estimated
cropsrowing
least half a
:e defoliation
sore effective
a time of that
on a life of
iimtion from
jungle

11 military
210:: of either
that it is an
"-33 0f che'mi- -
i that it is an
ilsruption of
sinus simply
?3 they were
?ment report,
1793 of crops
4 guerrillas,
gave a
I mild and
L"It'erous to

?ring seven
 
thirties)
MOIOs'ical
Tm'ld War,
nt of the
"Motion of

.- ..

.n ..-

each type of material. Of these materials. Agent Orange,

 

 

109

since 1963. their use had risan two hundred and seventy-one percent?more
than double the rate of increase in the use of pesticides, though pesticides are
still far more extensively used. By 1960, an area equivalent to more than three
per cent of the entire United States was being sprayed each year with herbi-
cides.

Considering the rapidly growing civilian use of these products. it is perhaps
not surprising that the defoliation operations in Vietnam escaped any signi?-
cant comment ln the press, and that the American public remained unaware of
the extent to which these uses had their origin in planning for chemical and
biological warfare. Nevertheless, between 1941 and the present, testing and
experimentation in the use of 2.4.5-T. and other herbicides as military
weapons were going forward very actively at Fort While homeowners
were using herbicidal mixtures to keep their lawus free of weeds, the military

were screening some twelve hundred compounds for their usefulness in biologi-

cal-warfare operations. The most promising of these compounds were test-
Sprayed on tropical vegetation in Puerto Rico and Thailand, and by the time
fullscale defoliation operations got under way in Vietnam the U.S. military
had settled on the use of four herbicidal spray materials there. These went
under the names Agent Orange, Agent Purple, Agent White,
designations derived from color-coded stripes girdling the shipping drums of
the most widely used
as a general defoliant. consists of a ?fty-?fty mixture of butyl esters and of
2.4-D and Agent Purple. which is interchangeable with Agent Orange.
consists of the same substances with slight molecular variations. Agent White,
which is used mostly for forest defoliation. is a combination of and
Picloram, produced by the Dow Chemical Company. Unlike 2.4-D or 
which, after application. is said to be decomposable by micro-organisms in soil
over a period of weeks or months (one field test of in this country
showed that signi?cant quantities persisted in soil for ninety-three days after
application). Picloram?whose use the Department of Agriculture has not
authorized in the cultivation of any American crop?is one of the most persist-
ent herbicides known. Dr. Arthur W. Galston. professor of biology at Yale. has
described Picloram as ?a herbicidal analog of and an article in a Dow
Chemical company publication called ?Down to Earth" reported that in ?eld
trials of Picloram in various California soils between eighty and ninety-six
and a half per cent of the substance remained in the soils four hundred and
sixty-seven days after application. (The rate at which Picloram decomposes in
tropical soils may, however, be higher.) Agent Blue consists of a solution of
cacodylic acid, a substance that contains ?fty-four per cent arsenic. and it is
used in Vietnam to destroy rice crops According to the authoritative ?Merck
Index," a source book on chemicals. this material is "poisonous." It can be
used on agricultural crops in this country only under certain restrictions
imposed by the Department of Agriculture. It is being used herbicidally on
Vietnamese rice ?elds at seven and a half times the concentration permitted
for weed-killing purposes in this country, and so far in Vietnam something like
?ve thousand tons is estimated to have been sprayed on paddles and vegetable
?elds.

Defoliation operations in Vietnam are carried out by a special ?ight and Agent Bloc.h 5'

12th Air Commando Squadron of the United States Air Force. from a base at 

Bien Hoa, just outside Saigon. with specially equipped 0?123 cargo planes.
Each of these aircraft has been ?tted out with tanks capable of holding a
thousand gallons. 0n defoliation missions. the herbicide carried in these tanks
is sprayed from an altitude of around a hundred and ?fty feet, under pres-
sure. from thirty-six nozzles on the wings and tail of the plane, and usually
several spray planes work in formation, laying down broad blankets of spray:
The normal crew of a military herbicidal-spray plane consists of a pilot. a col-
pilot, and a technician, who sits in the tail area and operates a console regu-
lating the spray. The equipment is calibrated to spray a thousand gallons of
herbicidal mixture at a rate that works out, when all goes well. to 'about three
gallons per acre. Spraying a thousand-gallon tankload takes ?ve minutes. In
an emergency. the tank can be emptied in thirty seconds?a fact that has par-
ticular signi?cance because of what has recently been learned about the nature
of at least one of the herbicidal substances.

The o?icial code name for the program is Operation Hades. but a more
friendly code name, Operation Ranch Band. is commonly used. In similar fash-

4' 

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President Johnson from twenty-two scientists. including seven Nobel laureates.
The petition pointed out that the "large-scale use of anticrop and ?noniethal'
antipersonnel chemical weapons in Vietnam" constituted ?dangerous precedent"
in chemical and biological warfare, and it asked the President to order it

stopped. Before the end of that year, Dr. Edsall and Dr. Matthew S. Meselson, 

a Harvard professor of biology. obtained the signatures of ?ve thousand scien-
tists to co-sponsor the petition. Despite these protests. the area covered by
defoliation operations in Vietnam in 1967 was double that covered in 1966. and
the acreage of crops destroyed was nearly doubled. .

These ?gures relate only to areas that were Sprayed intentionally. There is
no known way of spraying an area with herbicides from the air in a really
accurate manner, because the material used is so highly volatile. especially
under tropical conditions, that even light wind drift can cause extensive
damage to foliage and crops outside the deliberately sprayed area. Crops are
so sensitive to the herbicidal spray that it can cause damage to ?elds and gar-
dens as much as ?fteen miles away from the target zone. Particularly severe
accidental damage is reported, from time to time, to so-called "friendly" crops
in the Corps area. which all but surrounds Saigon and extends in a rough
Square from the coastline to the Cambodian border. Most of the spraying in
Corps is now done in War Zones 0 and D. which are classi?ed as free ?re
zones, where. as one American of?cial has put it. "everything that mOves in
Zones and is considered Charlie." A press dispatch from Saigon in 1967
quoted another American oihcial as saying that every Vietnamese farmer in
that corps art-a knew of the defoliation program and disapproved of it. Dr.
Galston. the Yale biologist. who is one of the most persistent .critics of Ameri-
can poiicy concerning herbicidal operations in Vietnam, recently said in an
interview. "We know that most of the truck crops grown along roads. canals.
and trails and formerly brought into Saigon have been essentially abandoned
because of the deliberate or inadvertent failing of these defoliant sprays:
many crops in the Saigon area are simply not being harvested." He also cited
reports that in some instances in which the inhabitants of Vietnamese villages
have been susln-cted of being Vietcong sympathizers the destruction of food
crops has brnuzht about complete abandonment of the villages. In 1986. herbi-
cidal old-rulinan caused extensive inadvertent damage, through wind drift, to a
Very large plantation northwest of Saigon owned by the Michelin
rubber inn-rests. As the result of claims made for this damage, the South Viet-
namoso authorities paid the corporate owners, through the American military.
nearly a million dollars. The extent of the known inadvertent damage to creps
in Vietnam can be inferred from the South Vietnamese budget?in reality. the
American military budget?for settling such claims. In 1961'. the budget for
this conuw-nsation ??as three million six hundred thousand dollars. This sum.
probably re?ects only the harest emergency claims of the people
affected. 3 .

According to Representative Richard D. McCarthy, a Democrat from upstate
New York who has been a strong critic of the program, the policy of allowing
applications for defoliation operations to ?ow. usually without question. from
the level of the South Vietnamese provincial or district chiefs has meant that
these local functionaries would order repeated sprayings of areas that they
had not visited in months. or even years. The thought that a Vietnamese dis-
trict chief can initiate such wholesale Spraying. in effect without much likeli-
llood of serious hindrance by American military advisers. is a disquieting one
to a number of biologists. Something that disquiets many of them even more is
what they believe the long-range effects of nine years of defoliation operations
will be on the ecology of South Vietnam. Dr. Galston. testifying recently
before a congressional subcommittee on chemical and biological warfare. made
these observations: 

?It has already been well documented that some kinds of plant associations .

subject to spray, especially by Agent Orange. containing 2.4-D and have.
been irreversibly damaged. I refer speci?cally to the mangrove associations
that line the estuaries. especially around the Saigon River. Up to a hundred
thousand acres of these mangroves have been sprayed. . . . Some (mangrove
areas) had been sprayed as early as 1961 and have shown no substantial signs
of recovery. . . . Ecologists have known for a long time that the mangroves
lining estuaries furnish one of the most important ecological niches for the
completion of the life cycle of certain shell?sh and migratory ?sh. If these
plant communities are not in a healthy state. secondary clients on the whole-

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113

latex production in the affected plantations fell oi! by an average of between

thirty-?ve and forty per cent. According to a report by the two scientists. 

large variety of garden crops were devastated in the seemingly endless number

or small villages scattered throughout the affected area. Virtually all of the . . .

local inhabitants . . . depend for their wellbeing upon their own local pro-

duce. These people saw their crops . . . literally wither before their eyes." The
Cambodian claim is still pending. .

Until the end or last year, the criticism by biologists of the dangers involved
in the use of herbicides centered on their use in what were increasingly con-
strued as biological-warfare operations, and on the disruptive effects of these
chemicals upon civilian populations and upon the ecology of the regions in which
they were used. Last year, however, certain biologists began to raise serious
questions on another Score?possible direct hazards to life from On
October 29th, as a result or these questions, a statement was publicly issued
by Dr. Lee DuBridge, President Nixon's science adviser. In summary, the
statement said that because a laboratory study of mice and rats that had been
given relatively high oral doses of in early stages of pregnancy
?showed a higher than expected number or deformities" in the the
government would, as a precautionary measure. undertake a series of coordi-
nated actions to restrict the use or in both domestic civilian applica-
tions and military herbicidal operations. The DuBridge statement identi?ed the
laboratory study as having been made by an organization called the Blonetics
Research Laboratories, in Bethesda. Maryland. but gave no details of either
the ?ndings or the data on which they were based. This absence of speci?c
information turned out to be characteristic 0! what has been made available to
the public concerning this particular research project. From the beginning. it
seems, there was an extraordinary reluctance to discuss details of the pur-
ported ill effects of on animals. Six weeks after the publication of the
DuBridge statement, a journalist who was attempting to obtain a copy of the
full report made by Bionetics and to discuss its details with some of the gov-
ernment of?cials concerned encountered hard going. At the Blonetics Laborato-
ries, an oii?lcial said that he couldn?t talk about the study, because "we're
under wraps to the National Institutes of government agency
that commissioned the study. Then, having been asked what the speci?c doses
of were that were said to have increased birth defects in the fetuses or
experimental animals, the Blonetics o?icial cut off discussion by saying.
?You?re asking sephisticated questions that as a layman you don?t have the
equipment to understand the answers to.? At the National Institutes of
Health, an of?cial who was asked for details of or a copy of the study on
2.4.5-T replied. ?The position I'm in is that I have been requested not to dis-
tribute this information." He did say, however, that a continuing evaluation of
the study was under way at the National Institute of Environmental Health
Sciences, at ReSearch Triangle Park, North Carolina. A telephone call to an
of?cer of this organization brought a response whose tone varied from wari-
ness of downright hostility and made it clear that the o?icial had no intention
of discussing details or results of the study with the press.

The Blonetics study on was part of a series carried out under con-?
tract to the National Cancer Institute, which is an arm of the National Insti-
tutes of Health, to investigate more than two hundred compounds. most of
them pesticides, in order to determine whether they induced cancer-causing
changes. fetus-deforming changes. or mutation-causing changes in experimental
animals. The contract was a large one. involving more than two and a half
million dollars' worth of research. and its primary purpose was to screen out
suSpicious-looking substances for further study. The ?rst visible fruits of the
Blonetics research were presented in March or last year before a convention of
the American Association for the Advancement of Science. in the form or a
study or possible carcinogenic properties of the ?fty-three compounds; the ?nd-

ings on 2,4,5-1? were that it did not appear to cause carcinogenic changes 'in 

the animals studied. .

By the time the report on the carcinogenic properties of the substances was
presented. the results or another part of the Blonetics studies. concerning the
teratogenic. or fetus-deforming. properties or the substances. were being com-
piled. but these results were not immediately made available to biologists out-
side the government. The data remainedmsomewhat frustratingly. in the view
of some scientists who had been most curious about the e?ects of herbicides?
out of sight. and a number ot?attempts by biologists who had heard about the.-

 
  
 

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115

Harvard. In early October, Miss Johnson?s friend, in a conversation with Pro-
fessor Matthew Mesalson, mentioned Miss Johnmn's report on the preliminary
Bionetics ?ndings. This was the ?rst that Dr. Meseiaon had heard of the exist-
ence of the Bionetics study. A. few days previously, he had received a call
from a scientist friend of his asking whether Dr. Meselson had heard of cer-
tain stories, originating with South Vietnamese journalists and other South
Vietnamese, of an unusual incidence of birth defects in South Vietnam, which
were alleged to be connected with defoliation operations there.

A few days later, after his friend sent him further information, Mr. Mesel-
son decided to obtain a copy of the Bionetics report, and he called up an
acquaintance in a government agency and asked for it. He was told that the
report was ?con?dential and classi?ed," and inaccessible to outsiders. Actually.
in addition to the preliminary report there were now in existence the full Bio-
netics report and a statistical summary prepared by the National Institute of
Environmental Health Sciences, and, by nagging various Washington friends,
Dr. Meselson obtained bootlegged copies of the two latest reports. What he
read seemed to him to have such serious implications that he got in touch
with acquaintances in the White House and also with someone in the Army to
alert them to the problems of in the hope that some new restriction
would be placed on its use. According to Dr. Meselson, the White House people
apparently didn't know. until that moment that the reports on the adverse
effects of 2,4,5-1? even existed. (Around thattime. according to a member of
Nader?s Raiders, "a tremendous lid was put on this thing" within government
agencies. and on the subject of the Bionetics work and ?people in gov-
ernment whom we?d been talking to freely for years just shut up and wouldn?t
say a word.") While Dr. Meselson awaited word on the matter, a colleague
of his informed the press about the ?ndings of the Bionetics report. Very shortly
thereafter. Dr. DuBridge made his public announcement of the proposed
restrictions on the use of -

In certain respects, the DuBridge announcement is a curious document. In
its approach to the facts about 2,4,5?1? that were set forth in the Bionetics

inn and military purposes, and the words in which it describes the reasons for
restricting its use appear to have been very carefully chosen:

"The actions to control the use of the chemical were taken as a result of
?ndings from a laboratory study conducted by Bionetics Research Laboratories
which indicated that of mice and rats given relatively large oral
doses of the herbicide during early stages of pregnancy showed a higher than
expected number of deformities.

?Although it seems improbable that any person could receive harmful
amounts of this chemical from any of the existing uses of and while
the relationships of these effects in laboratory animals to effects in man are
not entirely clear at this time. the actions taken will assure safety of the
public while further evidence is being sought."

These actions, according to the statement, included decisions that the
Department of Agriculture would cancel manufacturers' registrations of 
for use on food crops, effective at the beginning of 1970, ?unless by that time
the Food and Drug Administration has found a basis for establishing a safe
legal tolerance in and on foods," and that the Departments of Agriculture and
the Interior, in their own programs, would stop the use of in populated
areas and in all other areas where residues of the substance could reach man.
As for military uses of the statement said. "The chemical is effective
in defoliating trees and shrubs and its use in South Vietnam has resulted in
reducing greatly the number of ambushes. thus saving lives." However, the
statement continued, ?the Department of Defense will [henceforth] restrict the
use of to areas remote from the population." .

All this sounds eminently fair and sensible, but whether it represents a

candid exposition of the facts about and the Bionetics report is debata- 

ble. The White House statement that the Bionetics ?ndings ?indicated that
of mice and rats given relatively large oral doses of the herbicide
during early stages of pregnancy showed a higher than expected number of
deformities" is, in the words of one eminent biologist who has studied the Bio?
netics data, "an understatement." He went on to say that "if the e?ects on
experimental animals are applicable to people it's a very sad and seriOus situa-

 

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117

their total number was, in fact, greater, not less, than this average. Including
controls but excluding litters, the total number of animals used in the 
studies was two hundred and twenty-?ve. Analysis of the results by the
National Institute of Environmental Health Sciences found them statistically
?signi?cant.? and this is the real purpose of such a study: it is meant to act
as a coarse screen to shake out of the data the larger lumps of bad news.
Such a study is usually incapable of shaking out anything smaller; another
kind of study is needed to do that. .

Thus, the DuBridge statement seems to give rise to this question: If the
Bionetics study, based on the effects of on two hundred and twenty-?ve
experimental animals of two species, appears to be less than conclusive, on the
ground that ?the study involved relatively small numbers of laboratory rats
and mice,? what is one to think of the adequacy of the tests that the manufac-
turers of pesticides make? it, as the DuBridge statement says, ?at best it is
di?icult to extrapolate results obtained with laboratory animals to man," what
is one to say of the protection that the government affords the consumer when
the results of tests of pesticidal substances on perhaps a hundred and twenty
rats are of?cially extrapolated to justify the use of the substances by a popu-
lation of two hundred million people?not to mention one to two million
unborn babies being carried in their mothers? womhs?

The very coarseness of the screen used in all these tests?that is, the rela-
tively small number of animhls involved?means that the bad news that shows
up in the data has to be taken with particular seriousness, because lesser
effects tend not to be demonstrable at all. The inadequacy of the scale on
which animal tests with, for instance, pesticides are currently being made in
this country to gain approval is further indicated by the fact that a
fetus-deforming en'ect. that might show up if a thousand test animals were
used is almost never picked up, since the studies are not conducted on that
scale; yet if the material being tested turned out to have the same eifect.

quantitatively, on human beings, this would mean that it would cause between-

ihree and four thousand malformed babies to be produced each year. The tera-
togenic effects of on experimental animals used by the Bionetics people.
however, were not on the order of one in a thousand. Even in the case of the
lowest oral dose given rats, they were on the order of one in three.

Again, it' is fair to say that what is applicable to rats in such tests may not
be applicable to human beings. But it is also fair to say that studies involving
rats are conducted not for the welfare of the rat kingdom but for the ultimate
protection of humanbeings. In the opinion of Dr. Epstein. the fact that the
used in the Bionetics study produced teratogenic effects 'in both mice
and rats underlines the seriousness of the study's implications. In the opinion
of Dr. McLaughlin, this is even further underlined by another circumstanCe?
that the rat, as a test animal, tends to be relatively resistant to teratogenic
effects of chemicals. For example, in the late nineteen-?fties, when thalido-
mide, that disastrousiy teratogenic compound, was being tested on rats in oral
dosages ranging from low to very high. no discernible fetus-deforming eifects
were produced. And Dr. McLaughlin says that as far as thalidomide tests on
rabbits were concerned. "You could give thalidomide to rabbits in oral doses at
between ?fty and two hundred times the comparable human level to show any
comparable teratogenic effects." In babies born to women who took thalido-
mide, whether in small or large dosages and whether in single or multiple dos-
ages, between the sixth and seventh weeks of pregnancy, the rate of deforma-
tion was estimated to be one in ten.

Because of the relatively coarse testing screen through which compounds
like pesticides?and food additives as well?are sifted before they are
approved for general or specialized use in this country, the Food and Drug
Administration theoretically maintains a policy of stipulating, as a safety
factor, that the maximum amount of such a substance allowable in the human
diet range from one two?thousandth to one one-hundredth of the highest
dosage level of the substance that produces no harmful eifects in ex
animals. (In the case of pesticides. the World Health Organizati takes a
more conservative view, considering one two-thousandth of the ?no-"effect" level
in animal studies to be a resaonsable safety level for human exposure.)
According to the standards of safety established by F.D.A. policy. then, no
human being anywhere should ever have been exposed to because in
the Bionetics study of rats every dosage level produced deformed fetuses. A
?no-eifect" level was never achieved.

 

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119

the thirty-second emergency-dumping procedure. Dr. Pfeiffer has recalled going
along as an observer on a United States defoliation mission last March, over
the Plain of Reeds area of Vietnam, near the Cambodian border, during which
the technician at the spray controls was unable to get the apparatus to work.
and thereupon dumped his whole load. ?This rained down a dose of 
that must have been fantastically concentrated.? Dr. Pfeiffer has said. ?It was
released on a very watery spot that looked like headwaters draining into the
Mekong River, which hundreds of thousands of people use? In another
instance. he has recalled. a pilot going over the area of the supposedly
"friendly" Catholic refugee villages of Ho Nai. near Bien Hoa, had serious

engine trouble and dumped his whole spray load of herbicide on or near the -

village. In such instance. the concentration of dumped upon an inhab-
ited area in Vietnam probably averaged about a hundred and thirty times the
c0ncentratlon reccommended by 2,4,5?1? manufacturers as both effective and
safe for use in the United States.

Theoretically, the dangers inherent in the use of should have been
removed by means of the steps promised in the White House announcement
last October. A. quick reading of the statement by Dr. DuBridge (who is also
the executive secretary of the President?s Environmental Quality Council) cer-
tainly seemed to convey the impression that from that day onward there
would be a change in Department of Defense policy on the use of in
Vietnam. just as there would be a change in the policies of the Departments
of Agriculture and the Interior on the domestic use of But did the
White House mean what it certainly seemed to be saying about: the future mil-
itary use of in Vietnam? The White House statement was issued on
October 29th. On October 30th, the Pentagon announced that no change would
be made in the policy governing the military use of in South Vietnam.
because?so the Washington Post reported on October 3Ist??the Defense
Department feels its present policy conforms to the new Presidential direc-
tive." The Post article went on:

Pentagon spokesman?s explanation of the policy, read at a morning press
brie?ng. differed markedly from the written version given reporters later.

"When the written statement was distributed, reporters were told not to use
the spokesman's [previous] comment that the defollant . . . is used against
enemy ?training and regroupment centers.?

?The statement was expunged after a reporter asked how use against such
centers conformed to the Defense Department?s stated policy of prohibiting its
use in 'pOpulated areas.? 

But the statement wasn't so easily expunged. A short time later. it was
made again. in essence, by Rear Admiral William E. Lemos. of the Policy
Plans and National Security Council Affairs Of?ce of the Department of
Defense, in testimony before a subcommittee of the House Foreign Affairs
Committee, the only dl?'erence being that the phrase "training and regroup-
lnent centers" became "enemy base camps." And in testifying that the military
Was mounting herbicidal operations on alleged enemy base camps Rear Admi-
ral Lemos said:

?We know . . . that the enemy will move from areas that have been prayed.
Therefore, enemy base camps or unit headquarters are sprayed in order to
make him move to avoid exposing himself to aerial observation." 

If one adds to the words ?enemy base camps" the expunged words "training

and regroupment centers"?centers that are unlikely to operate without an.
population?what the Defense Department seems;

accompanying civilian
actually to be indicating is that the ?areas remote from the population"
against which the United States is conducting military herbicidal operations
are ?remote from the population" at least in part because of these operations.

As for the Bionetics findings on
mental animals. the Department of Defense indicated that it put little stock in
the dangers suggested by the report. A reporter for the Yale Daily News who
telephoned the Pentagon during the ?rst week in December to inquire about
the Defense Department's attitude toward its use of in the light of the
Bionetics report was assured that ?there is no cause for alarm about defol-
lants." A week or so later, he received a letter from the Directorate for
Defense Information at the Pentagon which described the Bionetics results as
based on ?evidence that when fed in large amounts to highly imbred
andsusceptible mice and rats. gave a higher incidence of birth defects than
was normal for these animals." After reading this letter. the Yale Daily New:

"16669 

 
  

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121

business of manufacturing It appears that the presence of a dioxin con-
taminant in the process of manufacturing is a constant problem among
all manufacturers. Three years ago, Dow was obliged to close down its 2.4.5?1?
plant in Midland. Michigan. for several months and partly rebuild it because
of what Dow people variously described as ?a problem" and ?an accident."
The problem?4r accident?was that workers exposed to the dioxin contami-
nant during the process or manufacture came down with an acute skin irrita-
tion known as cblor-acne. The Dow people. who speak with considerable pride
of their toxicological work (?We established our toxicology lab the year Ralph
Nader was born." a Dow public-relations man said recently, showing, at any
rate. that Dow is keenly aware of Nader and his career), say that the chlor-
acne problem has long since been cleared up. and that the current level of the
dioxin contaminant in Dow's 2.4.5-T is less than one part per million, as
Opposed to the dioxin level in the used in the Bionetics study. which is
alleged to have been between ?fteen and thirty parts per million. A scientist
at the DuBridge o?ice. which has become a coordinating agency for informa-
tion having to do with the question, says that the used by Bio-
netics was ?probably representative" of 2.4.5-T being used in this country?and
presumany in Vietnam?at the time it was obtained but that considerably less
of the contaminant is present in the 2.4.5?1? now being produced. Evidently. the
degree of dioxin contamination present in varies from manutacturer to
manufacturer. What degree 01" contamination high or low. was present in the
quantities of shipped to South Vietnam at various times this spokesman
didn't seem to know.

The point about the dioxin contamination of 2.4.51]? is an extremely impor-
tant one. been use if the suspicions of the Dow people are correct and the cause
of the fetus deformities cited in the Bionetics study is not the but the
dioxin contaminant. then this contaminant may be among the most teratogenl-
cally powerful agents ever known. Dr. McLaughlin has calculated that it the
dioxin preSent in the Bionetics 2.4.5-T was indeed responsible for the terato-
genic effects on the experimental animals. it looks as though the contaminant
would have to be at least ten thousand times more teratogenicaily active in
rats than thalidomide was found to be in rabbits. Furthermore. it raises
alarming questions about the prevalence of the dioxin material in our environ-
ment. It appears that under high heat the dioxin material can be produced in
a whole class or chemical substances known as trichlorophenols and pentachlor-
ophenois. These substances include components at certain fatty acids used in
detergents and in animal feed. -

As a consequence of. studies that have been made or the deaths at millions
of young chicks in this country after the chicks had eaten certain kinds oi:
chicken teed. government scientists are now seriously speculating on the possi-
bility that the deaths were at the end or a chain that began with the-spraying
of corn crops with 2.4.5-T. The hypothesis is that residues of dioxin present in
the remained in the harvested corn and were concentrated into certain
byproducts that were then sold to manufacturers of chicken feed. and that the
dioxin became absorbed into the system of the young chicks. One particularly
disquieting sign of the potential or the dioxin material is the fact that bio-
assays made on chick embryos in another study revealed that all the embryos
were killed by one twenty-millionth ot a gram of dioxin per egg. 

Perhaps an even more disquieting speculation about the dioxin is that 2.4.5-T
may not be the only material in which it appears. Among the compounds

that. several experienced biologists and toxicologists suspect might contain or 

produce dioxin are the trichlorophenols and pentachlorophenols, which are
rather widely presunt in the environment in various forms. For example. a
number of the trichlorOphenols and pentachlorophenols are used as slime-kill-

ing agents in paper-pulp manufacture, and are present in a wide range of con-?

sumer products. including adhesives. water-based and oil-based paints. var?
?nishes and lacquers. and paper and paper coatings They are used to preterit
slime in pasteurizers and fungus on vats in breweries and are also used in
hair shampoo. Along with the 2.4.5-T used in the Bionetics study. one trichlor-
ophenol and one pentachloropbenoi were tested without teratogenic results.
But Dr. McLaughlin points out that since there are many such commands put
out by various companies. these particular samples might turn out to be ?by
the reasoning of the allegation that the 2.4.5-1? used by Bionetics was unu-
sually dirty?unusually clean.

56MB

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123

There have been a number of reports from Vietnam both of animal abortions
and of malformed human babies that are thought to have resnlted tron: spray-
ing operations in which was used. But such scattered reports, however
well tounded, cannot really shed much more light on the situation. The fact is
that even in this country. the best-fed, richest, and certainly most statistics-
tuinded of all countria on earth, the standards for testing materials that are
put into the environment, into drugs, and into the human diet. are grossly
inadequate. The screening system is so course that. as a teratology panel of
the )irah Commission warned recently, in connection with thalidomide, "the
teratogenicity of thalidOmide might have been missed had it not produced mal-
formations rarely encOuntered." In other words, had it not been for the fact
that very unusual and particularly terrible malformations appeared in an
ubvious pattern?for example, similarly malformed babies in the same hospital
at about the same limb-pregnant women might still be using thalidomide. and
lesser deformations would, so to speak, disappear into the general statistical
background. As for:more subtle effects. such as brain damage and damage to
the central-nervous system, they would probably never show up as such at alL
if such risks existed under orderly, normal mediml conditions in a highly
developed country, how is one ever to measure the harm that might be done to
unborn children in rural Vietnam. in the midst of the malnutrition, the dis-
ease, the trauma, the poverty, and the general shambles of war?

or 
New York, March 5. 1970.

The EDITORS,

The New Yorker

Dun Sites: In an article that appeared in The New Yorker on February 7th, I
wrote that Dr. Lee DuBridge, the President's science adviser, iSSued a state-
ment last October at the White House saying that bemuse a laboratory study
had shown a ?higher than expected number of deformities" in the fetuses of
mice and rats exposed to the herbicide agencies of the United States
gm?ernment would take action to ratrict the use of that substance in this
country and in Vietnam, where it. was being used in extensive military defolia-
tion Operations. This action, Dr. DuBridge announced, would include the can-
cellation, by January is: of this year, or Department of Agriculture permits
for the use of on some American food crops unless the Food and Drug
Administration bad by then been able to determine a safe concentration of the
herbicide in foods. Dr. DuEridge further announced that the Department of
Defense would thenceforth ?restrict the use of to areas remote from
the population" in 1Vietnam. His statement added that these actions and others
"Will assure the safety of the public while further evidence [of the alleged
harmful e?'ects of is being sought."

Four months have passed. and 2,4,5?1? is still being used as widely as ever.
The Department of Agriculture has yet to cancel its permits for the use of the
herbicide on food crops in this country, and the Department of Defense is con-
tiuuing to use it in poliulated areas of Vietnam. In the meantime, o?icials of
the Dow Chemical Company, which is one of the largest producers of 
have been maintaining that the samples of used in the study cited by
Dr. DuBridge, which was done by the Bionetics Research Laboratories, of
Bethesda, Maryland, were uncharacteristic ot' the currently being pro-
duced, because the material tested by Bionetics?which did not come from
Dow?was contaminated to an unusual extent by a toxic substance identi?ed
as symmetrical This contaminant, usually
called dioxin, was alleged by the Dow people to be present in the Bionetics
samples at a concentration of approximately twenty-seven parts per million,
and they claim that the that Dow is currently producing contains the
dioxin contaminant in concentrations of less than one part per million. The
Dow people maintain that their currently produced does not appear to
have the effect of deforming rat fetuses. in January, a Dow of?cial told the
Department of Health, Education, and Welfare, "We strongly urge that action
concerning the status of he held in abeyance until [Dow's] testing Pro-
gram is completed [in] April." The United States government?s failure so far
to place the promised restrictions on the use of in this country may in
Dart be attributed to this plea.

45?362 

 

MUG

uf-ttoszt I

'166?1

inh?.

2693

Toxicology of Chlorinated Dihenlzo-p-dioxins
by M. Schwetz,? LII. Iorris,? 6.1. Soarsoho,? Vii. Rowe; Gehrig:

Ll; tillerson,T and 0.6. Eerbigi

Severe toxicological responses have been

associated with certain chlorodibenzodioxins.?

One of these responses is chloracne. a folli-
culosis ?rst associated with skin contamina-
tion by chlorohydrocarbons in 1899 (1),
Serious outbreaks of chloracne-like lesions

associated with runaway reactions in the?

production of 2,4,5-trichlorophenol occurred
in Germany in the early 1950?s (2). 2,4,5-
Trichlorophenol itself does not cause acne
(3), but the contaminants which may be
formed in the uncontrolled production of
are extremely potent
acnegens (2). 2,3,7,8-Tetrachlorodibenzo-p-
dioxin and tri- and tetrachlorodibenzofuran
were isolated from the contaminants formed
in 2,4,5-trichlorophenol production and were
demonstrated to be strongly positive acne-
gens when applied to rabbit ears (3). By
using the rabbit ear test, the acnegenic
potency of 
xin was con?rmed in 1962
(4). In addition, is extremely
toxic in the chick embryo assay (5) and is
highly embryotoxie in rats Another
chlorodibenzodioxin, hexachloro'dibenzo-vp-
dioxin (HCDD), is known to be positive for
the chick edema factor, a condition char?
acterized by hydropericardium, ascites, and
anasarca (5, 7).

?Chemical Biology Research, The Dow Chemical
Co.. Midland. Michigan 48640.

fHuman Health Research and Development Cen-
ter. The Dow Chemical Co.r Zionsville. Indiana
46077. 

September 1973 CORRECTION 

the term mg or mg/kg-day

.2.

II
coo:
(.0

. NJQD.
Experimental  .1
Materials '3

The chlorodibenzodioxin samples used in
these studies are identi?ed and described in
Table 1. Studies were limited in some cases
by availability of pure samples.

Acute Lethality

Samples of 
hexachlo-
rodibenzo-?p-dioxin, and octachlorodibenzo-p-
dioxin were evaluated for acute oral lethality
in several animals as summarized in Table 2.

Test materials were administered as sus-
pensions in corn oil or as corn oil: acetone
solutions in single doses by gavage.
The animals were deprived of feed for 16 hr
before dosing. Afteridosing, they were ob-
served for signs of toxicity including body
weight changes for two to eight weeks.

Lethality of via skin ab-
sorption was tested on rabbits of mixed
sexes with doses of 31.6, 63, 126, 252, and
500 ,ug/kg body weight. The compound was
applied as a 0.01% solution in acetone to
the abdominal skin which had been shorn.
After the acetone evaporated, the trunk of
each rabbit was wrapped in cotton to pre-
vent ingestion. The rabbits were housed in
individual holding cages and were observed
for signs of toxicity including body weight
changes for three weeks.

Parenteral lethality was determined by
injecting rabbits of mixed sexes intraperi?

87

on pp 89-97 should read ug or mg/kg/day.

'16672

toneally with 31.6, 63, 126, 252 and 500
Jug/kg of as a 0.01% corn oil
suspension; control rabbits were injected
with corn oil. The rabbits were housed in
individual holding cages and were observed
for signs of toxicity for four weeks. The
were calculated by the Weil modi-
?cation of the Thompson method (8, .9) or
by the Litch?eld and Wilcoxon method (10).
The acute lethality studies were terminated
when it was evident that the survivors were
not showing signs of toxicity.

Irritation

Rabbit eyes were examined priorto ex-
periments and found to be free from defects
or irritation. Approximately 2 mg of 2,7-
DCDD, 2.3.7.8-TCDD, HCDD, or OCDD were
instilled in the conjunctiva] sac of one eye;
the contralateral served as a control.
The eyes were examined at various times

after treatment for conj unctival redness and-

chemosis, iritis, and corneal injury. Re-
sponses were categorized according to in-
tensity.

Rabbit Ear Bioassay For Acnegenic Activity

_Acnegenic activity of 1,23,4-
TCDD, HCDD, and OCDD
was tested by applying 0.1 ml of either a
solvent solution or the supernatant of a
solvent suspension of each compound to the
inner surface of the rabbit?s ears ?ve days
a week for four weeks. The ears were ex-
amined weekly for signs of chloracne, in-
flammation and hyperkeratosis. The re-
sponses were divided into ?ve categories:
(1) none, (2) very slight, (3) slight, (4)
moderate, and (5) severe.

Responses in the ?rst three categories in-
clude no response to mild irritation, in-
creased ear thickness, slight enlargement of
the follicular aperture, slight exfoliation
and slight crust formation. These responses
alone are not considered indicative of chlor?
acnegenic activity. Categories 4 and 5 are
indicative of acnegenic response and are
characterized by comedo formation, in-
creased ear thickness and hyperkeratosis.

September 1973

Teratology

Pregnant adult Sprague-Dawley (Spartan
strain) female rats weighing approximately
250 were used to study teratogenicity of
the chlorinated dibenzo-p-dioxins. The day
sperm were ?rst present in a vaginal smear
was considered day zero of pregnancy. The
animals were housed individually in wire-
bottom cages in a room controlled for tem-
perature, humidity, light cycle and noise.

Commercial laboratory rat chow and water'

were provided with choice.

Corn oil: acetone solutions with
varying amounts of test material were given?.
in 2.5 ml/kg dosages by gavage. Dosages
were calculated using daily body weights.
Rats were treated with 100 mg of 2.7-.
DCDD/kg-day, 0.1; 1.0, 10, or 100 pg

?HCDD/kg-day and 100 or 500 mg 

kg-day on days 6 through 15 of gestation.
Control rats received 2.5 ml/kg of corn oil:
acetone 9:1) orally. All rats were observed
daily throughout pregnancy and were
weighed on days 6, 13, and 21 of gestation.
Pregnant females were sacri?ced by carbon
dioxide anesthesia on day 21 of gestation:
the uterine horns were exteriorized through
a midline incision in the abdominal wall.
and the number and position of live. dead.
and resorbed fetuses were noted. After be-
ing weighed and sexed, the fetuses were
examined for external anomalies; the crown-
rump length was measured with a vernier
caliper. Half of each litter was preserved in
Bouin?s solution and later examined for soft
tissue anomalies the other half was
preserved in alcohol. cleared and stained
with Alizarin Red?S, and examined for
skeletal abnormalities (12).

A 2 2 contingency table was used to
evaluate the frequency of anomalies and
resorptions within the fetal population and
between litters. Body weight and body mea?
su rements were statistically analyzed by an
analysis of variance and Tukey?s test (1.3).
In all cases, the level of signi?cance was


Chick Bioassay for Chick Edema Factor
The bioassay for chick edema factor was

89

?16673

01141;? 

Table 3. Lethality of 

 

 

Time of Number
death, days deaths/
Route of postadminis- . Dose, number
Species and sex Sample administration tration LDao, mg/kg mg/kg treated
Rat, male Oral 9?27 0.022 0.008 0/5
0.016 0/5
0.032 10/10 1
0.063 5/5
Rat, female Oral 13?43 0.045 (0.030?0.066) 0
Guinea pig, male Oral 5-34 0.0006 (00004-00009) 0
Guinea pig, male Oral 9?42 0.0021 (00015?00030)
Rabbit, mixed Oral 6?39 0.115 (0038-41345) 3
Skin 12-22 0.275 ((1142?0531)
intraperitoneal 6?23 0.032 0/5 Lu
0.063 2/5
0.126 2/5 
0.252 2/5 
0.500 3/5 45
Dogs. male Oral 9?15 0.30 0/2 \x
3.00 2/2
Dogs, female Oral 0.03 0/2 
0.10 0/2

 

Responses to individual doses are given in those cases in which an LDso could not be calculated. The
LDso for oral administration to rabbits was calculated by using the method of Litch?eld and Wilcoxon
the remaining values were calculated by using the Weil modi?cation of the method of Thompson

(15, 
I?Letters refer to sample identi?cation in Table l.

ient pain and conjunctival inflammation,
initially. Treatment with was
associated with delayed conjunctival chemo-
sis 13-22 days later. By day 27, the chemosis
had subsided, but the rim of the eyelid was
thickened and encrusted. In rabbits treated

with HCDD, the rim of the eyelid was en- 

crusted 27 days after treatment. Neither
corneal injury nor iritis was observed in any

of the animals following instillation of the 7

chlorodibenzodioxins in the conjunctival sac.

Acnegenic Response

Both and HCDD produced
acne in the rabbit ear bioassay as indicated
by the formation of comedones. Solutions of
(sample c) in benzene rang-
ing in concentration from 0.04 to 400 ml
produced a positive response with severity
increasing with concentration. A negative
responSe was obtained with a solution of
0.004 Jug/ml. In contrast, a chloroform solu-
tion of 50 ,ug/ml, did not
produce a positive response. With HCDD

September 1973

(samples a, b, c, and a response was
produced by solutions of 10 to 50 ag/ml in
chloroform and dimethoxyethane. Chloro-
form extracts from 10% suspensions of 2,7 -
DCDD or OCDD were negative, indicating
that these have a low?order or possibly no
acnegenicactivity.

Teratogenicity

The effects of chlorodibenzodioxins on
maternal and fetal body measurements, in-
cidence of fetal resorptions and anomalies
are given in Tables 4 and 5.

Rats treated With 100 mg/kg-
day on days 6 through 15 of gestation gained
more weight during pregnancy than
controls but showed no toxicity. There was no
eli?ect on fetal body measurements, or in-
cidence of resorptions, or gross. soft tissue
or skeletal anomalies.

HCDD. Administration of 0.1?100 pg HC-
DDI?kg-day was associated with a dose-
related decrease in maternal weight?gain

91

?16674



81.61 Jaqmaadas

96

Table 4. E??ect of treatment with chlorinated dibenzo-p-dioxin on maternal and fetal body measurements and the incidence of fetal resorption.

 

 

 

 

 

Maternal weight gain, Fetal Fetal Fetal resorptions, 
Test compound No. of body weight, crown?rump 

(sample) litters Days 6?13 Days 13?21 Days 6?21 g? length, mm Population Litter 
Control 13'? 8 5.68 1 0.05 44.5 1 0.1 7 22/337) 47 (14/30)
2,7?Dichlorodibenzo?p?dioxin 

100.0 mg/kg-day 5.80 1 0.09 44Hexachlorodibenzo-p-dioxin 

0.1 ,ug/kg-day 5.73 1 0.04 43.8 1 0.1 5 10/217) 47 9/19)
1.0 ,ug/kg?day 5.93 1 0.16 45.7 1 0.5 9 20/213) 74 (14/19)

10.0 ag/kg?day '5.12 1 0.05' 42.6 1 0.2' 25' 57/229) 94? (17/18)

100.0 ag/kg-day 3.65 1 0.28' 35.2 1 0.7' 85? (194/227) 100' (19/19)
Octachlorodibenzo-p?dioxin 

100.0 mg/kg?day 5.73 1 0.09 43.6 1 0.4 8 11/131) 42 5/12)

500.0 mg/kg?day 5.69 1 0.05 44.5 1 0.2 5 9/199) 41 7/17)

Sample identi?ed in Table 1; administered on days 6?15 of gestation as a corn oil: acetone solution.

..Mean 1 S.E. for various gestation times.
?Mean of litter means 1 S.E.
(number resorptions/number implantations).

(number litters with at least one resorption/number litters).
'Signi?cantly different from control by an analysis of variance and Tukey?s test (measurements) or the 2 2 contingency table (resorp~

tions) (0.05.

- 0519122 M00 -

ELGI Jaqmawdas

Table 6. Results of chick edema body weight, food consumption. and pericardial ?uid volume calculations of chicks treated with
. chlorodioxins.

 

Pericardial ?uid volume Positive for chick edema

factor based on

 

Body weight, g"

 

 

 

92991

 

Food Mean log - Calculated
Treatment (sample)' 11 Day 0 Day 21 consumption, 3? ml 1 SE. (100 ml) value Calculations Gross lesions


17.4 0.16 0.02 1.1717 

0.01 ug/kg 10 44 1 196 7 16.7 0.14 i 0.02 1.1181 -0.74 No No

0.10 Jug/17.2 0.19 :t 0.01 1.2688 +1.69 No No

1.0 pg/kg33.7 2.34 i 0.08 2.3680 +213 No Yes

10.0 pg/kg? 9 42 i 1 No survivors 11.2 1.29 0.62 1.5661 +1.47 No Yes

Hexachlorodibenzo-p?dioxin I

0 #g/kg 9 38 1 194 2 ?17.4 0.15 :t 0.01 1.1771 

0.1 ug/ke 10 42 1 197 4 17.8 0.11 1002 0.9978 ?1.93 No No

1.0 pig/18.2 0.09 0.01 0.9337 ?4.57 No No

10.0 pg/kg? 9 38 1 187 7 16.7 0.81 0.01 1.7294 +3.82 No Yes
100.0 pg/kg? 10 36 1 1 No survivors 13.3 0.62 0.24 1.5650 +2.72 No Yes

Octachlorodibenzo-p-dioxin (Day 20)

0% of diet 12 45 :t 1 141 8 13.3 0.08 i 0.01 0.8053 
0.1% of diet 11 45 1 124 :t 5 9.5 0.06 0.01 0.7839 ?0.21 No No
0.5% of diet 11 43 1 196 at 10 10.9 0.09 .t 0.01 0.9002 +0.91 No No

Sample identi?ed in Table 1.
?Mean 1 SE.
Grams/chick/day.

?Administered orally as a corn oil: acetone solution.

'Animals died on days treatment.
Animals died on days treatment.

One animal died on day 19 of treatment.

I'Animnlu died on days treatment.
Fed in the diet E: 100 mg/kg. 0.5% =5 500 rug/kg).

732/92 

Microscopic examination of this organ re-
vealed a highly variable pattern and degree
of hepatic necrosis with various degrees of
degeneration and regeneration of the hepato-
cytes, depending upon the post-treatment in-
terval. Necrosis was observed both in the
centrilobular and periportal areas. The de-
gree of necrosis of the liver was not suf?cient
to conclude that it was responsible for death.
Hepatic lesions were observed in rats, mice,
rabbits, and dogs. In addition to hepatic
involvement, other changes observed sporad-
ically include fat necrosis, periarteritis, ser-
ous atrophy of fat, and ascites.

Discussion and Summary

The studies reported here con?rmed the
high toxicity of In addition,
some perspective of the relative toxicities
of HCDD, and OCDD has been
obtained. and OCDD failed to
cause death in female rats given oral doses.
of 1 g/kg; even larger doses were given to
mice without causing death. Limited data
suggest that oral doses of approximately
100 mg/kg of HCDD are needed to cause
death in male rats. In the teratology study,
no deaths occurred following administration
of 100 ,ug,?kg of HCDD to female rats for 10
consecutive days.

is much more toxic than
the other chlorodibenzodioxins studied; the
ranged from 0.6 ,ug/kg in male guinea
pigs to 115 pg/kg in rabbits. Dogs appear to
be less sensitive than rabbits. Others have
reported 100% mortality in rabbits treated
with 10 ,ug/kg (15) and chick embryos
treated with 0.05 pg/egg (5 

Death following treatment with a lethal
dose of often delayed for
several weeks. Among the animals which
died following treatment, approximately half
the deaths occurred between 13 and 18 days
after treatment, with one animal dying as
late as 43 days after a single oral dose. In
mice and rabbits, there is a marked in-
dividual difference in susceptibility to this
compound which makes it dif?cult to con-
duct acute lethality studies.

If the results of the rabbit irritation

September 1973

test can be extrapolated to man, accidental
contact of these chlorodibenzodioxins with

the eyes should not present a serious threat

to vision. However, repeated contact with
the skin of small amounts of either 2,3,7,8-
TCDD or HCDD may be expected to produce
chloracne. Sensitivity was a
recognized by industry years ago, and prey-O
cautions have been taken to minimize its'?
occurrence and prevent contamination of:W
worker's skin. HCDD is apparently a less?fpa
potent acnegen than 2,3,7  

As previously reported, is
highly embryotoxic (6). The no-effect level
for embryotoxicity was 0.03 ag/kg?day of
In contrast to the high em-
bryotoxicity of the symmetrical 2,337,8-
TCDD, was not embryotoxic at
doses as high as 800 ng/kg-day (16).

By previously described de?nitions of ter?
atogenicity and embryotoxicity (17), HCDD
is teratogenic in the rat at a 100 ag/kg?day
dose level, given orally on days 6 through 15
of gestation. Treatment of pregnant rats
with HCDD caused embryotoxicity evid?
enced by a dose-related decrease in fetal
body weight and crown?rump length and
an increase in the incidence of fetal resorp-
tions (Table 4). Likewise, the incidence of
certain soft tissue and skeletal anomalies
increased in a manner related to the dose
level of HCDD (Table 5). A 0.1 ng/kg-day
dosage of HCDD had no effect on embryonal
or fetal development.

OCDD caused embryotoxicity but was not
teratogenic at 500 mg/kg?day. OCDD and
caused neither teratogenicity' nor
embryotoxicity at 100 mg/kg-day. Khera
and Ruddick reported that the ad-
ministration of 2 mg was
associated with microscopic myocardial and
pericardial lesions in rat fetuses. However,
examination of sections of myocardium and
pericardium from fetuses of dams- treated
with 100 mg doses in this study revealed no
morphological differences from controls.

Both and HCDD give posi-
tive results in chick edema bioassays (Table
6). This HCDD result is consistent with a
previous report that the HCDD isolated

waste

97.-

16677

16. Khera, K. S., and Ruddick, J. A. Polychioro-
dibenzo-p-dioxins: Perinatal effects and domi-
nant lethal test. in Wistar rats. Advan. Chem.

1?7. Schwetz, B. A., Sparschu, G. L.. and Gehring,

P. J. The effect of and esters of on

rat embryonal, foetal and neonatal growth and

Ser. 121, R. F. Gould, Ed., American Chemical development. Food Cosmet. Toxicol. 9: 801

Society, Washington, D.C., in press. (1971).
8}
CD 
CD

CO
(ID
0'3

September 1973

'165?5_

2694


q. 2/ 

0

Formation of Dibenzorlioxins and Other 
aBonrlonsation Products from g?
'Dlrlorinated Phenols and Derivatives 353

by ll.ll. langer,? LP. Brady,? and M. Briggs?

Introduction

hlorodioxins can be formed in a two-step
condensation reaction from ortho-substituted
chlorophenoxy radicals or anions (1-3). The

route is of signi?cance only where
strongly oxidizing conditions exist such as
a reaction of chlorine with pentachlorophe-
nol at elevated temperature. For the second
route we have investigated the condensation
of alkali metal salts of chlorinated phenols
which occurs spontaneously when these metal
salts are heated to temperatures above 
Since this reaction is strongly exother-
mic, it proceeds to completion in a very nar-
row temperature range once initiated.

For bimolecular reactions involving ortho
chlorines of both phenate molecules, dioxins
would be formed according to, the scheme
shown in eq. 1) as one of the possible reac-
tion pathways. If these phenates contain
halogen substituents in meta or para posi-
tions, these also are involved in the conden-
sation reactions and the product distribu-
tion depends on a variety of factors such as
(1) the total number of halogen substitu-
ents which determines the ease of removal
of halogen; (2) the arrangement of the mole-
cules within a if this reaction takes
place in the solid state, which in turn is in-
?uenced by the metal cation involved; (3)

 

Sastern Research Laboratory, Dow Chemical
USA, Wayland, Massachusetts 01778.

September 1973

steric effects which may facilitate the at-
tack by the second molecule at a particular
carbon or may prevent a nucleophilic attack
at a particular site; (4) electronic effects
which in this case allows halogen atoms in
meta positions to be removed more readily
than those in the other ring positions (4).

Experimental

Thermal reactions were carried out in bulk
as described below or in a differential ther?
mal analysis cell using a DuPont 900 Di?'er-
ential Thermal Analyzer.

Residues and sublimates were analyzed by
mass spectrometry on an A.E.I. MS-12 in-

strument coupled to a com- . 
puter using. a program called MASH (5). .

Results

The results of our extensive studies are
summarized in Table 1, which contains all
the pertinent information on this compli-
cated reaction system and with this the an-
swers to many questions regarding dioxin
formation.

From electronic considerations Cl atoms I

marked with an asterisk and located in
meta positions are more likely to be re-
moved than any of the others (4). The ?rst
two reactions occur in the solid state where
the molecules?anions as well as alkali metal
cations?are locked into positions which may
have an important effect on the reaction

Table l. Pyrolysis products from Na chlorophenates by mass spectrometry.

 

 

 

Rela-
tive
inten-
sity, Approx.
Compound m/a Species yield, 
O- 0
Sodium pentachloro- 456 100 Dioxin (8 c1) so 0 I c; 
phenat? 684 2 Trimer (12 Cl) 
. I
912 0.3 Tetramer ((I)
Sodium 388 100 Dioxin (6 Cl) 50 0? to
chlorophenate 58 Cl I (13
(solid) 2 3 Turner (9 c1) 
Cl'  
Cl 
0
Sodium 2,4,5- 320 100 Dioxin (4 Cl) <1 0? Cl
trichloro henate 
(at mp) 36-356 30 Dlmeru (5 Cl)
480 4 Trimers (6 CI) 1,
ci-
Cl 3
Stadium 2,4,6- 320 100 Dioxin (trichloro henate .
(amp) 9 430 33 Turner (3 CI)
640 50 Tetramer (8 Cl) -
800 30 Pentamer (13 Cl) Cl -
Sodium 2.4- 252 100 Dioxin (2 on <3 0? C1
dichlorophenate . 
(at mp) 378 10 Turner (3 CI)
504 1 Tetramer (4 Cl)
Cl
Sodium 252 100 Dioxin (2 CI) 7
(when) 378 10 Trimer (3 Cl) 0
414 20 Trimer (4 Cl)  

504 16 Tetramer (4 CI) Cl Cl
540 12 Tetramer (5 Cl)

I

September 1973

c1/\ 01/ C1

C1 C1

REFERENCES

1. Kulka, M. Octahaiogenodibenzc~p-dioxins. Can.
J. Chem. 39: 1937 (1961).

2. Tomita, M., Ueda, S. L, and Narisada, M. Diben-
zo-p-dioxin derivatives. XXVII. of
polyhalcdibenzo-p-dioxin. Yakugaku Zasshi 79:
186 (1959).

3. Pohland, A. E., and Yang, G. C. Preparation and
characterization of chlorinated dibenzo-p-dioxins.
J. Agr. Food Chem. 20: 1093 (1972).

4. Weingarten, H. Ullman condensation. J. Org.
Chem. 29: 977 (1964).

5. Briggs, P., Dix, D. T., Glover, D., and Kleinman,

September 1973

etc. (2)
R. A'multiinstrument data acquisition system for
use with mass spectrometry. Am. Lab. 4: 57
(Sept. 1972).

. Buu-Hoi, N. P., at 9.1., Preparation, propriet?s et
identi?cation de la ?dioxine? (tetrachloro-2,3,7,8?
dibenzo-p-dioxine) dans les pyrolysats de defoli-
ants a base d?acide trichloro-2,4,5-phenoxy-aceti-
que, et de ses esters et des vegetaux contam- .
inn?s. C. R. Acad. Sci. (Paris) Ser. 273: 708
(1971).

. Boer, F. P., et 31., The and molecular
structure of 
Acta 28: 1023 (1972).

:7
.
. O.

FESTIMONY OF A. T. TALCOTT 9?ri - 55?7-.-.-- . 3'34! frTalcott, administrator of Regulations and Labeling in the

Quality Assurance Department of The Dow Chemical Company. I have a Bachelor
of Science degree in chemistry from Alma College, Alma, Michigan, and have
done graduate work at Michigan State University but have no advanced degree.
I joined The Dow Chemical Company, Midland, Michigan, in 1957 as a chemist
in their analytical laboratories and moved through a series of technical
promotions in the analytical chemical field. In 1965 I moved to the Labeling/
Product Registration section of the Legal Department, becoming Supervisor

of that section in 1975. In 1976 I became manager of Product Safety Compliance
in Dow's Quality Assurance Department, which included the functions of

labeling, material safety data sheets, and product registration. In 1979

I assumed my present position where I am reSponsible for labeling policies on 
Dow products.

I am a member of the American Conference on Chemical Labeling, the Labeling
Standard Revision Committee of the Chemical Manufacturers Association, a member
or chairman of several committees dealing with regulations and labeling within
the Chemical Specialties Manufacturers Association, a former member of the.
Labeling and Precautionary Information Committee of the Chemical Manufacturers
Association, and the Labeling Committee of the National Paint and Coatings.
Association. I was also one of two industry representatives participating with
EPA to put together their National Symposium on Pesticide Labeling in 1974:

I will testify on the history and procedures used in the development of 
labels by The Dow Chemical Company and to the registration requirements for__
labeling used by the Pesticide Registration Division of EPA which was paTtr'

of the USDA in 1968 prior to the formation of EPA. I will also testify -..-..
n. . Q?irlu? ?ff' 



-- 4 ?66582.-



09969le M00.

7-1-1

..

(riuicbaauudia, Specialists, and legal and regulatory counsel

in areas_pertinent to such a product.



The labeling used on KURON weed and brush killer in the?time?period?1967?
wank

through 1969 was in?part specified in regulations for pesticides which were called

. vuq 
economic poisons under the statute in effect.at?thatntimei Thesei equired the 1'
product to bear a label containing the name of the product, the name and address of
the manufacturer or registrant, the net contents, an ingredient statement which
was prescribed in a Special manner, a warning or caution statement as appropriate

to regulations prescribing same, and directions for use??which if complied with,

would be adequate for the protection of the public. These elements of a label were

'further prescribed in a certain manner by the regulations and interpretations

promulgated by the Pesticide Registration Division and required a preclearance or
acceptance by that Division before they could be used on the package of the
pesticide. This labeling was also submitted to the state of Arizona and other
states for review as part of the state registration of the product required prior
to sale.

The product name KURON, the company name, and certain other elements such
as the registration number required to appear on the label are for-the mos:
part self-explanatory.

For precautionary labeling purposes,pesticides were placed in one of four

dimomuk
different categories based on certain toxicity data that would be in ani al tests

Jimmy? at?
provided to the Pesticide Registration Divisions These were cali?dfcategories

one, two, three, and four; qidh category one being materials that were highly

toxic and requiring the word "poison" and the skull and crossbones to be used on

*Trademark of The Dow Chemical Company

-- uachut'LBS were ten-

fold decreases in toxicity from the lower-numbered category.

During this time precautionary statements used on pesticides were Set



forth in a document called "Interpretation 18, Revision 2" published in the



Federal Register of March 9, 1962 as an interpretation of 7 CFR. Part 362:

 

There were two ways in which this interpretation was used.. One related to
signal words and statements of hazard that were used on pesticides/bontaining

specifically-listed active ingredients and the othertpADp set forth general
procedures based on the toxicity categories previously mentioned which was

applicable to KURON weed and brush killer.
. ,xcLu-I?Jd?a??! 2" .  .
Applying the principles stated in-Interpretation 18, one comes up with the

precautionary labeling which was accepted by the government as adeguate to protect



the public:and used by Dow on.its product KURON weed and brush killer.~l

--



.. 

Quoting from section "Labels of products in the third

.

category should carry the word "caution" in statements indicating the means of

avoiding the principal hazards of use. Use of the skull and cross bones, the

and antidote statements are not necessary for these products.?

wordm?poison,f
Another part of the label which may need more explanation is the ingredient

statement. Ingredient statements~are required to be made on all pesticide products

and must be stated in certain ways. The constituents of a product must be

characterized as either active or inert ingredients. The active ingredients must

be described by their chemical name.

The two ways in which ingredient statements can be listed re Specified
t??.uwu 7 kid 0da,mjtu?443 a. 
in section 362.7 of the-regulations. ?These+w?re to list each active ingredient

and its percentage and the total of the inert ingredients or to list the total
of the active ingredients and each individual active ingredient in descending

order of percentage without the actual percentage of each ingredient.p1us the} 

'f

wmm-esrvaazz: TEL. 

this product. Such a listing would not have been acceptable from the government'S'
standpoint since this is not an active pesticidal constithent; additionally, the
initials TCDD do not represent a common chemical name_acceptable to the federal
gowernment so such listing would again be prohibited. The other reason for not
listing TCDD as an ingredient is that the limits of analytical detection at the
time of production of the KURON in question were such that TCDD was not found in
this product.

There are other sections of the'label that are, of course, pertinent to
the allegations made concerning the product KURON weed and brush killer. Among
these are the instructions under a section titled fWarning" that "applications
by airplane, ground rigs, and hand di5pensers should be carried out only when
there is no hazard from drift." Other instructions which are meaningful to a
user of such products are the instructions that "coarse sprays are less likely
to drift? as well as the_instructions on application rates and times for specific
applications of the product.

The procedures followed for labeling pesticidal products are an application
of the principles that are included for labeling of chemicals set forth in the
American National Standard for Precautionary Labeling ANSI ZlZ9.l.? This is a
consensus standard which was not published until 1976, the
benefit of review of such organizations as the American Society of Agricultural

Engineers, the National Safety Council, the American Society of Safety Engineers,

the Society of Toxicology, the American Conference of Governmental

-7. ..

13885

 

.. 

organizations with an interest in conveying precautionary information\nn.1.

Industrial Hygienists. and numerous other government, industry, and independent,

These principles prescribe a signal word, statement of hazard, precautionary
measures, and as appropriate, instructions in case of contact or exposure as
procedures for providing precautionary labeling. The labeling used by Dow on

KURON weed and brush killer demonstrates these principles.

Human! 

The procedures used within Dow to develop labeling in l967-involved the
initial labeling being developed from a use standpoint by a technical Specialist
from our Agricultural Products Department along with the individual responsible 
for submitting registration applications to the federal government, and
precautionary information coming from a labeling specialist like myself. This
draft copy then would be typed and sent for review by the numerous specialists
within the Company with input on the health and safety aspects coming from
toxicologists, industrial hygienists, medical doctors, individuals from our
quality function in connection with our manufacturing people providing input on?
the composition of the product, trademark and patent comments coming from our
attorneys, and the original three persons reviewing these inputs and those of

can-'-

our marketing personnel. Once these inputs had been composited,.the labeling

It' a

was sent to the federal government for their acceptance or suggestions for change
if they?did?not accept it.

Upon return of a copy of the accepted labeling, the labeling specialist
would initiate the actions responsible for getting the labels or the preprinted
container copy to our production people for application on containers of the
final product.

It is important to remember when developing or reviewing labeling for a
product that the labeling must be based on the product as a.whole; consequently,
the labeling must be viewed in its entirety, not just in parts?-since it is the.

whole product that is used, not a part.

I have compared the labeling used by other pesticide producers of similar

(33 products and find the-labeling used, which also was required to be submitted

to the Pesticide Registration Division before being used, to be essentially the

same where applicable as that used on the product KURON weed and brush killer.
Based on my education, training, and experience, it is my opinion that the

label used on the product KURON weed and brush killer in the time period 196T

99969171 M00

_through 1969 met or exceeded all regulations and standards of care pertaining
to such products; and further the directions and precautions if followed were
adequate to prevent accident or injury with the handling and application of

the product.

?*16687

 '2702

 an d} e?fcat?Qu-W
V?daa?agemen

 *"Val'mlaoezszoul


.c - 
png?; ism?l?Ap;  I. l_

I 156688 
- 'l'i\l. t- 

ll ult- II by?: 

Hi-tx (?Ih A

The current status of
phenoxy herbicides


In January of 1971 a review of "The Status
of 2.4-0. 2.4.5-T. Silva; and 
cides" was-reported in DOWN TO 
(Vol. 26. No. 4). It now seems desirable and
benefiCiat to repeat pertinent parts or that
paper as well as to include additional data
and information generated since then

This new reView concerns itself with 2.4- 
l2.4.-dichlorophenoxyacetic aCid). 2.4.5-T
l2.4.5-ttichlorophenoxyacetic acrd). Silvex
l2-i2.4.5-trichlorophenoxy) propionic aCidt
and MCPA l2?methyl-4-chloronhenoxy
acetic acrd). While these herbiCides appear
to be quite similar in structure. each com-
pound exhibits unique characteristics 
respect to selectivity for broadleaf weed
and,'or brush control. The formulation at
these compounds into specific esters 0i
amines further contributes to SpeCies se-
lectivity as well as to partiCUlar techniques
lor spray preparation and application

The phenoxy herbicides have been uti-
lized commerCiatly for twenty years Or more
Without harm to humans or and
were registered on the basis of satety tests
conSidered adequate by USDA at the 

In 1966 the USDA and FDA. on a
Study conducted by the National 
of Sciences. declared that the concepts til
"no-residue? and "zero lulriruntm" HflJISIfi'i'
hen til pesticides on tuorl crops were no
lnnper tenable and residue tolerances would
have to be established to enable 
tuttistrillton under these lit (irtiui
to accomplish tit-s. addilirintil lixiirity tinil
sludies, It; itii-
rI-til Ililvt: [mutt by
USDA .iitli FDA Puhln Ilh- H'Ill 

. tr
~r

I  

'1  

MOO

now been transferred to the Environmental
Protection Agency (EPA) Scientific srurlies
subsequent to April 1966, when the USDA
abolished the no resrdue status. have shown
the phenoxy herbiCides to represent nu
hazard to man and his envrronment wher-
property applied in acc0rdance With label
recommendations. The i55ue oi 2.4.5-T 0n
food c: ops is still undergoing reView by EPA.
primarily at the inSistence of envuoii-
mental gr0ups and as an outgrowth of its
previous usage in Viet Nam. It is extremely
important to recognize that industrial. far
any or range/and usage of 2.4.5-7' are not
alfected by the Current contraversy. The
2.4.5-T i53ue has become one of prinCipIe
rather than monetary value to the manu-
lacturers and it has been in this context
that Dow has continually pur3ued the sci-
entilic rome to resolve the potential 
0t to man and his 
til 

Development of the phenoxy heiliicidw
in the mid-forties resulted in three 
herbicides for right-ot-way. fOrestry and.
other industrial markets These prOt'ltit?t:
(2.4-0. 2,4 S-T and silveiti me not 
by patents Seven (IOntlanClili 
were one or more (Ii die--
than: products and 0t MCPA ilt ti"-
wheit USDA annmincurl iit 
IIU tes'irlue status un tllHi li-i-.i .-
nii April 13 1966 Tithal: Di.i--
DOW MUIIHJIIHI Rhllii'J 
sun and H.iyw.it-i Tllr'\.
tti Ftir- l' l=i'
Hl'llilt Ttilt'litlt- ii? Th"

it?l'tl'lv :li I. 7  

jtssas

 Tub}. ?my. Ll r-1 in; . 
puritirms by 300 mull-u.- ?MP-Hunts iinrl iln- 'ltu'r-
tolerant." for each iiurhiizirle Ilii: opportunity to present tht: premium:

I range or pasture grasses and other data relatian in theJudgment entered by mice that the law requires to be struck Im- 
Court of Apmais. District of tween bent-lit and risk Hearing to be held .

iItron [ohrevrcwi?gtp Itrv??r. $531.
I Harrison Weliford. et al (petitioner?7,7. . 1-: - 34;. )4 .rkl-waning egg?igrstiauegfge - - .fi?etmooeraeaigiizlaf?yt- -

..

fragistrationsiofzr -- - .I . I
1 manu'a' 'mmumpons {of ust 6 order should remain in eiiect in spite   
use around the home. recreation areas . .

 and sim?at sues I I  oi Doyvs _oinwaetIionsI liq the oridezr..thereiiy  
.. - -  lation .- .3.981193  . e, 

2 2 4 5 Tusas on om?uo?s'mamad rice. Ruckelshaus states that the hearing

should proceed "forthwith"

October-Nader'pbuWQN Task Force sub-
mits containing 
.additionai data residuesinmeatand meat I

?v

.
In"!  
Hair-#23?

H. a .
"-511 :?ri

for human consumption. 

The case is remanded to EPA adminis-
trator "for further Consideration and for a
decision supported by a reasoned analysis

 oi the 

Ir - --: - nl-I?Hi  
January 197k Ruckeishausannomces 
3 60 day review beginning January 18. All - - .. .  
interestedrgroups and individuals areIin- . . 3.5.3349! 
vited toI any additional --  

  --   -  data??hr. 9n 

    

   
  
    
   
 
 
  
 
  

   

r.  - -
     
 lgqag?ug?g! %%Ms2.4.5-1? is or is notIanf imminent hazard to -  
Lf?fT' 3" 5?  the White.  4-. for-thquPb-yc
 realm Seientific Advisery?Com-' 55?- 

   

   
   
   
 

 
 
 



 i *fzi "ii-31'; 

- r- bi agnappg leg I   agar.214??-T?dvisow   

1- we report'Isubmitted _gq?gj -.

  

'r

 

but not released at this "m . -- .. 7 
JUN 23. Advrsory Com2.4-0 in apples. 
mittee report released by EPA. Committee bane {mugsleho?s 03?s: manu?sn  
recommendsi'hv? restoring regis- mats? tolerant; has $5 =in 1_ :53"
tration o! status airisting?pri'been estabiis ed}  in asparagus .
to April 1970_wrth.loliowrng - from use of pnrya?the I Qdium salt. .I In 
 . with the peti; 
.0 .. .. . I tions .196] is expected to be  I
- 0' 99?? TCDD 0" ?'S?mg sufficient i?g?ggl?-?Dzln blueberries. crar.-' 

 

. arid 0-1 PW berrie "and" I for silver: inappies
 and in grass in- 
tied analysz being furnished by man-' u? cmd?s?ghm" im?m' previous work and from 
-. reSiduai?samtife's analyzed by Dow in 197m  I I-- -
l3) formulations around home hear wafn-"-: Addit?mhaI-Work-Supponed by the Task  
-, -  mg tat-possibleIdanger to' pregnant fr': Fer-eh?" gmdimcass?tm?wue data  

5




rm - I. women'a .animais,,exposure'rtg?lan 
  a - air?  .    :azal 'at..thi= af- awaan 
 4)cond 'Ittphengxf' . 
. food chain magnification --  -.  . ascrops and .95 new rm?. 
a I003 i305-t ??Omo" mg phenol in'fthe' animal tissues and mill. 

m9 advels" ?Heats Residue analyses? indicate - 01 f5? 

I r; EPA issues a purpOrtotl a? phenoxy acids in m0? mod ?1th 
etermination and Order or12.4.5.T which ?me of harvest. Negug'me msuhm low I

?ames-2 .- grit}: It were PM 1.95?ng lg .v  
1 

 
      

 

nit-u: level in April 1967 lot
hilld? grains

Feeding studies in Harry Won.- um-
rlucted for each plienmy carnpound at levels
at 30. 100 and 300 for two weeks and
at 1000 for three weeks in the total
analv.

       
 

 
    

  
 
 

  

      
 
 
   

 
   
  
 

 

 
 

  
  

when by Butane-tin.? 

rmtiltml that 2 4 5 .Il virty hIqh (hunt-p.-

lr:vels wtis iprmlumiui ttlul
turmerl in uurzr- and rats Subse-
quent studies showed that a tone 
nant. 2.3. 7.8 - telrachlomrlilmnzo - p-dioxin
responsable tor some, _1

I

     
  

 
 

 
 
   
   
 
 
       
 

    
   

 

 .  F?s-ti i  ?r u-
 -      
contained: 27  
   are} -.
. . .tnl?mrapum  
 "l .-  stews .weakti?  214:9 ?Preduceg, 29.. a  
   r?striction?ltsm. high! 1 .- i. .
essa'ry'toPdairy Cattle graziri' and?i'n rabbits atdose?s'as hit} 735 "lib, 
treated with 2.4-0. silver: or MCPA day. The obvious concern is to produce - It
The phenoxies were also led to beef cattle without the centaminant. I  
and sheep at levels up to 2000 'in__th_ TCDD can be formed inth .nia?n  
total diet't?or 'four weeks. 2.4.5-trichlorophenblf  
then slaughtered immediately or one week 2.4.5-T. The conditions required jot: its 
alter withdrawal from a diet containing 2000 formation are high temperatures in . theitrg. 
herbicide Based upon the 300 ppm' presence of base. conditions 
- tolerance requested in/on grass from appli- occur in the alkaline hydrolysis of 7.37 .
cation 0! phenoxies up tbzpoundsber octet-13%tetrachlorooenzene x.  - 
no grazing reStriction should be'necessarff'jfj'ni No detectable dioit'in's have been fotgd 55'  -
for or MCPA The current limitation: This is due to the fact that 
for 2.4.5-T approved by EPAf'shoulqsulfice .hcursor;2.4;gichlorophenol is'fn't?ade by 
for both silyex and 2.4.5-T. This stavtesi'fDojlf: chlorinatiqn'l'of phen'  

 021.: animal 
treated area within two?wee ?th 

cationif'fii (Figure 1 shows typical Ere
on/in .?grassliirom 2.4.5-T aeoliqat 

ds at}! 


.. .

- 

I
If 
:re?ere a'pplied than}? the pagi
. - enox 'tbicide a tied.   
    a
Negligible?jfeisidue' tolerances can'lbe 
tamed. based on .90 day.
feeding 'studies in two speciesof ?amrn?als. -
Howey?ernt?oler?ances at higher _tpennissible) 
reSidue  leiels_' frequire Moi-year? leading  

 

-

remdgugiletswdies in ratsza 
the phdn'qx'y? herbicides warey'e'lireloped.
these long-.term leading studiesiw?re' not
massif i cathese .cqnp?uf?q?.? ?were

Welf?E'ohducted iti?r'ifvea??gftead 
studies}? 
studies iwi



two-19m: readinestudies with-silver.  

not reproduction ortertility studies:   
7 'day'leedih??st'udieshave 
-  "atisl?f'doiis: 24.51%? I  
Monique-Eton! errect' iev?latla?i?  .
Tabl'e'?zia?rbg?ui?r 1_ I  
Based on single oral doses  
2.4.5-T. siltrex anrl MCPA are" Classed as
toxic" values ranging
from 300 to 700 ?HI-'in hotly weight
.3 75. @045}th 
11111  

(II:

we:

   

  

 

siduq'rei?

  

studios in?ate 'and dogs: plus fertility and 

?iigiirgduci ion; ?agellum;   
Dow .




 
  

. . 

a

   

  
 

  

?title?

. - 
Ir.-

  

  
   
 
 
  
     
    
 
 
 
       
   
    
   
 
     
 

 

 

  
   
 
  
  
 
 
  
 
 
  
  
 
  
     

 
   

.u:52: vii-tresrrigls

41?

 .x'r   
?gial?t'nrw': cl'  

- 

 

.ng.

 

 

 


etime .

   
    
   

 

wesns 'Arre'n APP



  
  
 
 
  

I
i Matt. I Cw.? art-J lustre: lor residue )tmdut the IIFPHT program
I

.
I 't I amt: urn:

 
  
  

S: .
I. 
 "1152 
.  
I .Irf

     
       
 
 

?lu-l?
I Ikaq?i? 
.r :iw_=I .
(V -
3? 35:" fig: -
)9 
1:31"Int . - 4- . I '37?JagInv??Eo?kbl??

       
      

To date. apatyticat rneth have been op; I
developed and validated to a sensittvity of plants nor__trenstocate_dIto other plat}; pertisfh
04 in 2.4.5-T- acid, With and it can be washer; 

. - -. dip-"x3 -
proper manufacturing controls, 2.4.54 can - Dr. KearneyIatso 55mg: I - IggtItIiaI,


5"

  
 
 
 



$990.3 or: . . . -
may}. .-.  -  I I ?1

   
  
 
  
  

 
 

"-jj??px If}: . . .-.- - 4- . 
on: TCDD-{hayq {wen-5:- .;excapt 1111!:thme

  past two y'ehrsgI'IIEgIrly Ifore. 
   
ma?ig??a that $19.9. -I 

      

    
 
  
    

     

       

  

- eff?ck??lg
I:  meeting 1 .- ..
 _26. Hmong??'?fopli? - .- .. 
of WidespreEH?U'se of'tho Phe?'oxv'Herbi?? Recent?n?eye  .  -..
cides and Pictoram." This paper was pub- C'Oud 
Iished in 'theSeptember 1. 1971.Iissue or is cIIsearinQJrade r?evyzg?um  
. BioScience wot-21.- No. 17) and concludes bar. 15. .1971 Dei?a?wfiwith the "s't?te?n?t?iit' that "Impurities c'ai't be-
- an important fa'ctormparticmarly the chloral - ., I   
dibenzo?p?o?qurpur theseI'can be Icon; 9? rum? '2  -
I I . trolled by proper madutacturing techniques?? 24- cm'e'? feg?sulfr?i"
  Photodegonipb's'ition of chtorinated dibenzo- 513"? 

r-53 of May 

?sued. a 

    
      

 
 
  
 
 

  

 p-dioxinSI?: I?'gbown by D. G. grow(Sciengea; 9mm. pp. 748-749. Ahgust 20..  
WWI-dioxins Was culr'?i'hated ina 0? ?(31.5
- . day  in sessions  the Pesti- ,imin? 

- cide Chemis'trgi?ivismtw at tha'Arnerican 9"

-  Chemical??sisi? mean-w held: .. - it) 
A ton 1971?: II @953?? an

I arch Reportstrata? - d-  
Immenls erh resper ,g91mSI Wydad   197lon?11Ithefollowt
-  .   Coo c.an

 

"The Agfiqitfiofal Research Senate suntf?? Ithtorinated compounds. as 
wuh particut.:r OIIHJIIGSIS on 2.4 S-T were "lem?mndum 7-53, I  
started an February 1970 at Beltsv-Ite Md . FEWHNUIJ IS JS Whit-'- 1
hr 0 team led by brochemtst Pluhp. Contammunun as 2.4 S-T

..- 3 Half - rig-ii]: 'anei; am 1? ?tifitr 

     

       

       

   

    

 
 

 



 

 I. I .

uh ..

ties. The standards of acceptance for tech-
nical lenoprop. in terms of TCDD content.
will be the same as for technical 2.4.5-T.

It is now proposed that for registration
purpose all 2.4.5-1? and fenoprop products
to be sold in Canada must have a declared
and guaranteed TCDD content in the tech-
nical material of less than 0.5 ppm. Also.
the saurce lie. the manufacturer and place)
of technical 2.4.5-T and fenoprop must be
identi?ed The quality control measures
used are also to be described. Finally. the
analytical method used to determine the
TCDD content must be fully described and
shown to be sensitive to a level of 0.5 
TCDD. 2.4.5-T and fenoprop products are
not to be registered for sale for 1972 unless
these conditions are met. These criteria will
in future be a part of the protocol for regis-
tration and re-registration of 2.4.5-T and
fenoprop products.

Adherence to these standards will justify
deletion of the following warnings from
2.4.5-T product labels:

ltt'Exposure of pregnant women by any

route of contact to 2.4.5-T may be
harmful to the 

l2) 'Direct exposure of some women to this

product may cause a health hazard.?

The council of the Society of Toxicology
in a letter to the editor of Scrence Not 174.
November 5. 1971; commented as follows
On 2.4.5-7:

"The 2.4.3-T controversy involves a funda-
mental issue in safety evaluation. The iSSue
lS Simply whether Or not demonstration of
a teramgenic effect in some speCIes of ani-
mal. at a dosage level far in excess of any
possrlile human exposwe constitutes scr-
entiltt: grounds for harming the chernital
A UfluquHLnUSls hehnvuin

the affirmative. but the overwhelming ma-
jority do not. The EPA screnttfic advisory
on 2.4.5-T. which included sev-
eral toxicologiSts and teratologists. specif-
ically endorsed the majority opinion. To
accuse the scientific advisory committee of
"bias" for defending a commonly held sci-
entific principle is unwarranted. To select
a committee of ten experts who might share
the opposite "bias" would be extremely
difficult if not impossible."

The USDA has issued two reports deal-
ing With the b' telits of phenoxy herbicides

Restricting the Use of Phenoxy Herbi-
cides?Costs to Farmers.

Agricultural Economic Report No. 194

Restricting the Use of 2.4.5-T: Costs to
Domestic Users

Agricultural Economic Report No. 199

The USDA states that prohibiting the use
of phenoxy herbicides "would cost the U. 
farmers an additional $290 million to mam-
tatn Current agricultural prod! chart. In addi-
tion. farmers and their families would have
to work 20 million more hours to control
the weeds wrthout these herbicrdes. For
this extra labor, the farmers would obtain
no additional income 

Over a million dollars has been expended
Over the past two years to prove the safety
of phenoxy herbicides to man and his en-
vnonntent. From a SCientific base the
phenoxy herbiCides can contribute econom-
ically. efftCIently. and safely in the luture for
the control of broadleavert weeds and brush
on food crops. posh-re. rangeland. and non-
cropland areas as they have for over 20
years 

. 

sews; 

16693

at.

C9 i  .

 and n_nppreciste your thoughtfulness. in remembering our
 -   ?anew-mm:  - 

if}; in?T19'55' subject comparnr communicated. with 
1 -I.. Gi?nudsn 01:3. 8. A. in Geneva concerning skin irritation

the: Griswold Executire Res./    
- - J. Penta Office J. R. Myers 27 a 9
Res. Lab. PBS 17119 iw

14311.5: Melohing, Dowicides - r. . 4 (37' 

. now GHIMICAL LIMITED



1

m? WA- ROAD
MICHIMI 

February 22, 1957

c. H. Boehringer Sohn 
22b Ingel?nein: In Rhein - - -, - .Vi??-hl

Referencegv Dr. Hey/B I f.  
Gentlemen: I I
Subject} rrenmtim or Trichlorophenol to Avoid Chloracne 

 7? Thanks Ive-r: much for 'your' istter of February 11' end the I  
attached description of the work you have done on the pre-

.2 i  3c! Trichlorophenmcetic Acidto svcid the for-

 mation excites-e and consequent. dermatitis to
your-ww?ncm. This work is otvery greet interest to 

.I11-.



I


I. I
I
i.

41". .IJelfFifi-J.? 

 

   

1-:  ElFrank - 1. I  -.

5 

pmwigtgz  3-5-7 -     
  Wye-5* - 

. cog33: 


I-- 153! .-



it: 

from 2,1145-Trichlarophenolzwhich was referred to us by the 7 
 New Jersey. On January 27, 1955 -
 we'urote'tto Boehringer enclosing at data. sheet describing the 
., - hszerdsidue to, toxicity-and precautions for sure 

 end 2 and answered seven specific  

.35; up. iencein  - .   -

.., b'..-, .Hit-"""t-FT-vr- -. 

   

 

   

DJ

F.
GE

089L219 mm.

STATEMENT OF

LEWIS A. SHADOPF

QQOZZONI

ON 
(TCDD)
IN
THE ENVIRONMENT

8/4/76

'Iss?g

I My name is Lewis Shadoff. My residence address is

3031 N. 11 Mile Road, Coleman, Michigan. I am an Analytical
Specialist of the Analytical Laboratires of the Dow Chemical
Company's Midland Division. I graduated from the Polytechnic
Institute of Brooklyn in 1961 with the B.S. degree in
chemistry. I then attended Kansas State University and was

awarded a in chemistry in 1966 having done my doctoral

1891.39 1

research in mass spectrometry. Since then, I have been
by Dow Chemical working in mass spectrometry. In my
present capacity, I have reSponsibility_for the Operation

and maintenance of three gas chromatograph-mass spectrometers

including a high resolution unit.

ANALYTICAL INVESTIGATION OF TCDD
I have been working on the analysis of TCDD since 1970
determining the purity of material (1) and the

TCDD content of ester products.

Since April, 1973, I have been involved in the
development of gas chromatography?mass spectrometry methods
for the determination of TCDD at the picogram level
(corresponding to the part per trillion level in biological

samples after sample preparation).

Gas chromatography is an analytical technique used to
separate a mixture into its component parts. The separated
components may then be analyzed by a mass spectrometer which
yields information about the atomic composition of these
components. This combination has been used as an extremely

sensitive and specific detection method for TCDD.

The concept of parts per trillion is difficult to envision. 

If it were salt we were analyzing, it would correspond to the

aisvas

t'2"

ability to detectcmuggrain of salt in an Olympic size-swimming

pool of fresh water by analyzing one shot-glass full of water.

 

 

E=i
We have searched for TCDD in the following samples by this gig
technique: 

03
ENVIRONMENTAL SAMPLES ANALYZED FOR THE PRESENCE OF TCDD :3
CD
NUMBER TCDD 
SAMPLE SOURCE SAMPLES CONTENT: LIMIT OF DETECTION
Human Milk Texas 6 N.D. Ave. 3 ppt, range
(ref. 2) Michigan 1 N.D. 1-6
Fish (2) Texas - 20 N.D. - Ave. 7 range 2-18
- -- Arkansas 20 N.D. 
Mud(2) Arkansas 2 N.D. 4 and 6 
Texas - 2 N.D. 3 
Water(2) Arkansas 2 N.D. Ave. 0.2 
TeXas - 2 N.D. 0.1 
ow's Mi1k(3) Oklahoma 25 N.D. 0.5 or 
Missouri
Arkansas
Beef Fat(4) Texas 24 21 N.D. Ave. 4, range 2?7
- Oklahoma 3 3-4 
Missouri
Rice(5) l9 Ngn. Ave. 4, range 2?7

(12 of them commercial)
Shrimp Japan N.D. 
Vietnam 3 N.D.

not detected at the detection limits listed.

The significance of low part per trillion TCDD content is not pre-
sently known. It is under study at the EPA, Dow Chemical, and IITRI.

The methodology and some of the results have been reported at 4
scientific meetings (2). Publication of the work is now pending in

qrnals whose articles are reviewed by scientific peers.

-. - .
The cow's milk was collected from cow?s grazing on grass known to 

have been treated with 



The fish, human milk, mud, and water were taken from
environments where had been used on rangeland and rice
fields. Those samples from Texas were from the San Angelo,
Texas Reservoir which is an impoundment of the North Concho
River and has its water shed large acreages that have been
sprayed with herbicides for the control of mesquite
(0.5 lbs/acre acid equivalent) and brush (3-4 lbs/-

acre acid equivalent). The samples from Arkansas

EESLZSI MM

were taken from a lZS-acre pond in the heart of the Arkansas
rice growing area near Grady, Arkansas. _This water is used

to flood the rice fields which have been sprayed with the
equivalent of 1.25 lb/acre of acid four to eight weeks
previously. Later, when the water is drawn off the fields it

it pumped back into the pond for reuSe. Water in the pond also
collects run-off from surrounding rice fields during the rainy
season and is supplemented by wells. -This cycle has been in use
(including the proper use of herbicide) for 18 years up
to the time of this study. Thus, these should represent likely

locations for the accumulation of TCDD if it occurs.

Twelve of the rice samples were collected from grocery
Stores throughout the United States. The rest were taken

from experimental plots.
The shrimp was supplied by the Kien Hoa Shrimp Company.
REFERENCES

l. W. W. Muelder and L. A. Shadoff, Adz. in Chem. Ser.
120, (1973).

 

2. R. A. Hummel and L. A. Shadoff, 170th National
Meeting of the American Chemical Society, Chicago, Illinois
.(1975). 



3. Mahle, N. H., Higgins, H. S., and Gertzendaner, M. 3.,
.TSearch For The Presence of 2,3,7,B-Tetrachloro-p-dioxin in

Bovine Milk," submitted to Bull, Environ. Cont. Toxic. for

publication.

4. Kocher, C. W., Mahle, N. H., Hummel, R. A., Shadoff,

L., and Getzendaner, M. E., "Search For The Presence of 2,3,

(TCDD) in Beef Fat," submitted

to Bull, Environ. Cont. and Toxic. for publication.

S. Jensen, D., Hummel, R. A., Madrid, Ester, and Turley,

June, "Search For 2,3,7,8-Tetrachloro-p-dioxin in Rice," to

be published.

P891439 I

2950

Aria M'uml. 

Frum lln? lustilulr, Shu-lelm, 

AND 
01: (IllLORlN-Yl?lil) ACIDS
IN 

133'

Eur! Erna

'l?hc dislrilndion, vliminniiun and mcluhulism in plunL limsucsa
of chlorinated acids hum: been the subject of
(sec mg. Audux 1964). The fun: of
phunuxy acids in {he :animnl however, has not been
so ihm'oughly iln'cslignlud. 

Ancm'ding In early studios acid
is in urine by man, dogs, ?rats and rabbits
(H'il?ums 195(1). The n? :md p-chlm'odcrivnlives ulsu were. 
in In! rt?nnli)?. without conjugnliun, in the rabbit (Lz'm-y
Lt'un'x? 19-17). On 130mlng :lnd ?mythic acids
10 rabbits, sunn- pin-noxyuucliu acid was deluded in urine,
:dong wilh lln' unchanged acids, indiculing nuptial {i-(axidulion.

:1 gruup of :d Univcrsily, 
in :1 smjc? of I'm-ding will: ?th: and using :1 gnu
chmmulngruphiu luclhud of analysis, found ihnl chlorinnh?d
acids CIA-I). MCPA and \wrc ctunpiclciy
climiuuh-d with 1th urine in intact form, as \an? chlorinated
:u-id v! rd. 1903 
1963; Hm'hc and. 1516-1 John nl. 196-1).
(Zhlurinutcd m-ids and \wrv
di'?l'?dt't] in HM: runn-n uf hul Mun? :unnunls of Mn:
,?S-uxidnliun prnduvis, rurrvr.pmsdiug{ :u'idu, \wrr
HIM) nrim'. rsh?r was 

in nriuv us Hw :u'id. Num' Mummy hvrhit'itlr" 

h} :1 i'rnm 

01")
NC.

NCO




 I

2

lo (-uultl'lw in (.?Iurl: rl HI. up-
]ilying lrm'ri' lln?. l'npitl J?l?nnl vliminnliun
nl' nl'ull)? mlminislm?ul in :1 sln'l'l).
Elli nl' :u'livily \x'us l?Xk?l'l?lt'll Willi llir urine. in hours, and
lilnml low-ls llruplu-ll ln llL'lH'\\? (1.05 11.13.51}. in 21 lmurs.

animal species (in nul lu 
lwon rupnl'lccl.

'l'lic pl'c?scnl rcsulls {rum slmlics ul' llu: llislri-
huliun :md climiuuliun ul' smm- acids in rules.
pigs, calves and chickens. 'l'lm were in
cunnoc'liun will: toxicity studies which will lw. 
(maria:qu I-n-m- mum. 



Mulrn?uls

mm'm'. Aqueous solulilm cunluiuing 20 mg-nl' pl-r ml,
.13 line. lrirllmuoluminc snll, by (liluliug :t lur-
. mulnlirm. I 

2.4-1) sail. solution containing 20 mg ul' per
ml, prepared by (lissulviug the. pure acid lilS?l'l'i'll) in slighl
excess of aqueous until adjusting pH In about 7.

(unimn Aquwus sululiuu 20 mg of per
ml, prupurwl by dissolving lln: :utitl (111.1). 155?4 57"(3) in sliglil
excess of aqueous and adjusting ln :llmul 7.

2,13-1) ash-r. limulsinn containing 30 mg ul' pci' ml, as llu:
hulyl c-slcl', pvt-pared by lmmugmiizing :1 furmulnliun, in
pclruluum sulx?rnl. with 

Animals .

Dulu gin-n fur Weight and My: n-er lu cumliliuns :ll slurl of


Albino ruls, Aulicimcs slruin, 180-422? g. lmlli 

Pigs, Swmlisli "1.:mlrus" lu'cml, Ell-~25 lag. 10-? ~12 wwks, cuslrulml
mull-s. :uul 

Calves, ?65 kg, li??S weeks, liulli scsos.

Chicks, ln'uilcr, mu: wu-k. mulc.

Chitin-us. While Jim-ml, (ISA-1.0 lzn? 3-1-10 ?mm-L's, lmih



50395. Hampshire ln?cud - leg, :ulull lH-ns.



Arhm'm'slruliuu. ?lasing animals \?cvn- 
Must-s gin-u umlly, in sluH'l rspl-rinu-nls slumnrh (MW.
in lung: t'xlu'l'llm'nlu ul' \x?ulL'r.

.?iumpi'iny. sl-lu-rll-ll linn- lwl?ni'l- :Iml timing".
sumple in lulu-s. in :ll

 MOO

viess

 a m-u-l; arli-i'y, in piggx' vrna rava cranialis,
iu [mm a jugular win am] in ('llit'lu?lm from a winngin. l'iaMHa
was sc-paralmi 

A (If ilu: animals .Hill?l'irif'l'tl al 
linu?. inh rvals, in}; aflvy antallu-lizing; 
(rals aml m' nu-lmmai smiilim (pigs). 'l'isxm's
\Vi-i'r fur ?rms aml hialulngiral 
lnmi l-Irm' HIGH) and sampli-s ci-llm'lcd for analysis. 
slm'cil a! unlil analyn-il.

Anamh'mi Mir-thud. 'i?lu- acids were
i?ul?ik?ll {mm lmuiy fluids. aml by min-n! ('xll?aciiun. xvparatcd
by and (plaulilalichv
arsi-ording; In a prc-
Vina-,1; (firm: 1960 Win-n lh'c pl'um-iluru was scalqd (lawn
in handlu samples. as small as. 1 g. The givc's lul'al crin-
rt-nlralinn (if chl:urnplu-noxyaCulii: acitls. Thu limit of 
nf nu-lhml is 0.1?0.2 pg of uf sample, with tlu- 

sample. ainnunls, huwm-ci', almni lug/g.


A. Plasma levels

1. Singlc? (10.90

a. 2,5-1) mm'm'. Groups of rals (mali- and feinalv), pigs, cal-
ves and aml Hampshire) were given
amim: sail, in the form of a loclinical fm'mulalion, as single
oral doses equivals'nl l0 mg of 2.4-1) kg limly ?right.
011 (image ion-125 male anzl Ill 5- -IO pigs aml
chic-lama and 2 cam-s \wrv nsml. Tiluml samplL-s were lain-n (in
rals :Iilui' killing) nl il-liuur (luring firsl 12 lmul's
and lass fi't?tpicnily, and plasma analyzi-cl fur Hal-l).
Plasma vunrcniraliun-linu- arc in Figs. 1?4. (In
atlililiun, a number of llu?: animals wore al ('m'lain 
in Ol'llCI? la [issue (liali'ilmiion, src? SL?CllUll C, livlnw.)

'l'lit? peak plasma (If was usually allaim-(l
within 2 lmm's aflvr dosing in cinch-us and williin 4 --7 hours in
?w mammalian species. In ?l-lu- Spm?ics (-xa'mim-il a (lose (if 100
Ina/kg gau- a pull?: lm?i-l of about 100- "200 gig/ml. The plasma
lm?vls fur fcmalv rals luv-Tr llIan Hum-fur llu: IllalL'S. 'l?liu
half-lifv \?alm's ul' in plar-lna, ralvulaicul lln' i'ni'ws
:tl'lvi' plulling mi a main, al?i' in 'l?alnlc .1.

ll. .mH. 'l'lu- mpurimralx \wn- I'i'pL-alwl lin-
Hill ul' 'l?i'n maiv lilih aml rain-s 

Ilwa-x c-l? mg]!

L..


if?!

?31889?" 

 

 

4
300? 
1. Rats
200-
c?
. ow 9
]00- O. ?00
53:0
u?h 
'ah?n-rg' no. ?L?w  
I I As!) I 
:3 2004



3; l00~

 

 

PLASMA 2.4CGIVCS
100.4 
i 501 ?Ava-m? I 1-:

 

 

 

 

o- 1  
200-? I 4. Chicken
Ioo??uff?go
IO 20 - so so

Time (hours)
I-?i gu 14?4. )?lusmu lands of 1-1-9 in diffcn-ni spvcios nth-r :1
single urn] dust: of 2,4-1) mm?nu um Ul' 200 mg . 'l?hc rur-
pigs, valves and chickens \x'cn- obtained will: 5 animals ?In
each It'w-l and MW I'm- with sunk-s and 10 
2?4 animals nn mn?h p?inl nf Iln- rurw.

.-

(tlu'vvs ?Muile :uul half-lift: \?nlucs fulmd (Table 1)

similar In uhluim-d will: 2,14) amino.

r. 2,5,5-7' uml'm'. 'l'rn nmlu s'nls mu! 2 pigs \Vrn- ?in-n singh- 

urnl dusts hr :1 3.1.5-31? :Hlailh' 5:1? (Hm Mtg/kg}. 'l'lw plasma
:Is :rs lln? [Hm-mu hnli' ?fr \?nhn-r.

?Hind. Wrn' silnilzn In Ulnar ulslniuk'il will; Un- 3,1-1) suns.

918893 M00

16in:

IL 2.54) (as-IN. A h?rhnirul furnlululiu-n Ilu- hulyi

 

 

 

 

l'Sll'l? nf was giwn as single urn] (Inn's-Hf Hm lug/Lg; in r;
111310 rnls'. -1 pigs and L'nh'vx. Husmn sunnph-s ww'v 
fur lulu} :15 in HIV l?l'l't't?tnngi thpm'iuu'nis. 'l'ypirul rurx'm 
in Fig. Guru's fur pigs; won- 
lhnm? fur rah. :md plasma half-hic- values are given 
in 'l'uhh- I.

.



.3 

0 0?0 Rats

- 200?1

V. Calves



4 100_.ll 

10 20' 30 50

Time 11:59:51
i 1't'  l'lusmu nf 2,4-1) in rats and sing/{c
ornl dmu: of burg/I (3101' (100 mg . Tlu- curves \Vul?c
uhluincd with I'Hhs on each 1min! of ?w (rm-w) and valves.

Tn 1. Plasma half-life values, in diffL-n-nl spurios, of phcnuxy-
drrix?nliw-s given :15 a singh? m?ul ?Ins-.0. uf HIO 

 

 

Compound SprL-ics half-[Ne
lmurs
umim- Hal, mule 
Rm, fwnulv 3(3311' 7.5 1: {3.8
Chicken 7.7 0.7
Hal, nmiu 3.5 2: 0.5
8 2 
., ester lint, mulv ::nnim- 5n? Hui, 1mm" 3 i 0.6

 

2. Hrpruh'd min:I'm'sh'uh'nu

:amin'v and rxirr \u-rv gin-n m. ?ml 
{fan :nul {11335. 
In hqu gruups llu- pluhmn lc'nl HI 15,-1-11 \x'ns .M'rl?n {u dcrlim-

:Iml llu- urinary t'M'i?t'liun {u illi?l'k'u?u? us L-xrmlalil'iul in

.1670!

 nu. .uun in Ill ll'l 
denote the urinary emu-entraliuus of 2,-l-tl.
Plasma .ugfml
Number (at 24 hrs. after last dosedoses .
Amine lzster



I?ll: no. It A

 

PigPit: nn. 9 A

3 35 (200) 230 190 155
4 315 2-10 
7 335 2.1 295
520 (90) 25 . 
23 (550) 7 (25m 

 

Table. 2 by pigs no. A and fl A. One animal in each group, how-
ever. did not tolerate the repeated :ulministralion; the plasma
levels continued rising during the experimental period, and signs
of poisoning gradually developed (pigs no.3 A and 15 A, 'I?ahle 2).
in an experiment with two hens, 2.4-1) amine (300 lug/kg
day) given. orally for 12 and 24 days, respeetively, and (luring;
the experimental periud the plasma level (at 3 hours after dosing)
dropped from 150 to 20 [Lg/ml, on the average.

D.- Btoorl cells

In in nitro experiments 2.4-1) was added, at two eom'entration
levels,- to horse hlood and the samples agitated at room tem-
perature for-ti hours and 24 hours, respeetivety. Then the. 
cells were separated hy centrifuging; for 30 minutes at 25tltl 
Washed hy suspending in two volumes of either fresh plasma or
physiological saline for 5 minutes and centrifuging for another
5 minutes and then hy diluting with two volumes of
water. For comparison some samples were washed in physio-
logieal saline three times. The levels found in plasma and
hlood eells are presented in 'l'ahle ll. . 

A small proportion of the ahoul 1 5'3 of the plasma
level, was found in lnlood eells washed onee. The proportion did
not ehaugie on prolonged eonlaet hetweeu ptar-ma and eells.
??shing with plasum reinuveui more of the 23,-l-l) from the Mood
rells than did with saline. lit'lleutt'ui'?smiling; wi-Eh saline

l'emHth {lei-ll aluhr-t I'ium tlu' rellm

SHIRE

 713.97

7

a II. In m'lru ilislrilmlinn nl' 2.4-1) lwle-i-n hm'sv plasma and

('rll\ al lwu rum-i-nlralinn low-ls. lit?rhl?lt analysis ('tlls

washml nnm- (5 Iain.) will: 11 x'nluml-s ul' l'I'L'sll plasma (A)

m' ul' phpinlngiral (ll) lln'i-c linit-s (15 min. ('Ill?ll linI(-) will:

2 ul' physiulugiral salim- (C). Vallu's gin?n means (If J'L'salls
- ilnplicalc cspi-rinu-nls.

- 

[lullnl

 

 

 

 

Time of Ill ?ug added 100 ?ng added
mnlavl per ml lilmiil liloml
 - I 
l'lasma culls Plasma cells
A A ll  
5 13 1.5 2.1 (0.1 135 ll 17 (0The rcsulls reported in Table 4 were obtained physio-
logical with rals rc'cciviag 'single oral doses of 2,-l-D
amine and 

.1 hi 1- In aim; dislrilmliun of 2.4-1) ral plasma and 
culls MIN a Magic oral (lose (if cilia-r 2,4-1) amine or 2.4-1) butyl 
(100 mg in ('llc?ll caste). lcrm'u analysis ?10 blood cells 
\vaslu-d fur 5 min. will] 3. volumes of physiulngii-al saline. Valucs given

:ll_'c llu- means (if frunl two animals.

 

- r/ 
'l'iim- aflcr' m: 

 

 

 

Amine Ester
hours 
Plasma Illnod cells Plasma lilnml (-clls
'J'issnos

(ii-naps nl' rals (male. audit-male), pigs all-ll c'laiulicns 
and New llampshirv} won: given singh' oral doses nl' 2,4?1) amino
(100- 200 mg; and al ilifl'i-ronl inlon'als 3 d1
aninn'ils l'rmn l'?l'll gnqu sacrifii?t'il and ?mum; and bully
fluids analym-(l l'm' 'l?lw vspx-rinu'nls \voi'v Iwrl'i-rzm'il in
('unm-i'liun llu- plasma ill-scriln?il almw. 'l'la-
samnnu'iac-il in 'l?alili-s a 

'l'lli- i-l'll't'l limalr ul' 

ul if. H) was. in hauling; rals {Inalr 



16710

 I: 5.

ink-mulls .?Int'l' M'Hyh? urn] ?In-w of 2.1-1) :mtim' mg
lim'h \?ulm' is the nf I?mulh: 
gin-n in 

2.4.11,

4 animals, wilh llu- rungv

(mash Weigh!)

'l'isnm- Hf in Mink: and h-muh- mix nl 



.-



c, moo

Ci?!
5"

.

0888

 

Timut' 6 huurs 24 Imurs
l-?cmnlos Males 

15? (1313-475) 7? 37) 2( Lil-? 3.0) 1.5 (LS -- Li)
Lin-r 90C (35 -I2) 5( 3 9 3 2.9 5.2)
250 (Emu-48.3) 27 (19 15 (IO 
Lung I-IU 75) '1 3 
Spin-n 30?) (j 

Skt'h-inl 23 29) 5 -- 2f) 2 3.1) 0.6 0.2? 1.3)

Ft'c rs

90)



 

0m: animal.

'l'a I Tissue of in pigs at different aflvr :1
singlr urn! (hm: 0f :nmim- (MIG m-r 

-r

Each value is [In-
mcun u! rvsuhs {rum 3 willy-{he rangt- given in bruclu-is.

 

lug/n (fresh weigh!)

 

 - 
6 hours 21 hours 48 hours 7'2. hunrs

l?lusmu 2H) ([35 -?230 55( 13 10 5 16 3 
Lin-r 115( -?-125 27( 13 33 3 4  7 
Kitllu'y 190(165 3(Lung ?107 20Spin?curl mum-2.51-? 4.53) 2 2.7) 1.0-: "1.2
?ruin 8 IS 3 2 3.5) 1.5 1.3) --

PHI 3( 1.2? 3.3) 2( 1.0? 2.1) 
Skin 45( 30 55 - --
Hih- 2(4.3)
lirim- h'?t 18? (H0 95 ((30 ?--135 'l'llyruiul 55 (33 -
Adm-lull 95( 80 



l?mu'l'vna - - 71 -- -- 
Salivary glmul till 45 - -
313ml

75.( n3 

unian

16711

'l'a I) I 7. in :il cliffL-rrnl :ifli-i-

:i si'uglc- m-ul iiusi- uf :unim- (200 mg: value is [In
nu?nu (if l'l'Slliih' [rum 3 ll :mimuis. Willi tlu- Humi- gin-II in hl?ut'lwls

 

fig,? (fresh wright)

 

Tiswc -
6 hours 24 [unit's 48 hours

(72 --13I 15 (i0 2i) 5 (2 

Lin-r (5i) 25 (20 33 3 

120 (80 -?-170 80 (53 ?120 7 

Lung (-10 40 (28 49 4 (2 

Sph'tSiil'h?iili muscic 3.5 4.2) 'i.5 1.9) 1.3 

Bruin 1.5( 2.1) 

FM 1.1 1.5) 

Egg, yolk 0.2( 0.3) 0.2 
(0.1 <0.1

(0.1

 

Rpm/rd m'ul I'm' 2 Pigs were gin-n 5m: p.p.1n.
of 2.4-1) amine in {cc-cl lwic-c daily and other 1000
11.1).m. (if 2.4-1) :unime in their drinking wnlu'. The pigs Wen- 
hours iilL? )Hbi fur ii'iL' Olin-r ink-rm! in-lwn-n
1:19.! (1050 unti? killing wax irrcguiur. Each value given is Ihc ?It'll? (if
rcsuils frum 3 animals, wilh range given in 

a 8. Tissue Few-h. (if in mix, pigs, chicks and Chicthix :tfh'l'

 

2.1-0, pig/g (in-ah 

 

Tissue - Hats
Pius Chicks Chitin-m.


l?luslnu HI -i '32 8 ~35 1'2. (3 10 (3 
Lin-r 25 (H1 3(1) --13) (i 3 8 (5 ??Hi 15 (5 
15(8-7-26Linn,r 3i} -l 2 (i 15 (ii ?--iil (5 
Siu'ivlni . .

7 3 2 1.33?- 3 1.3 1:9) 3 3.5)
Skin 3n) 
lh'niu  - 2 2J3) 
1-1.1 . 2) .- m3 - 2.2)  (0.5 - 
1.5 (0.5 2.1)



.. 

thus :mimui.

0.1! ii. i I

IZCSSEMOG

16712

IO

pigs, (ln'uili'r) :Iml ('liit'lu-ns (Li'glnn'n). The pigs
in] liil?i, ?'liil' pqmn. of 2.4-1) :unim- :Hidl'ii,
:1 tiny. and lln- (Illn-r animals were gin-n :nninc, :il
low-l nl' 1o.]i.nl., in lln?ir drinking; Animals \u-i'u
sm?J'il'icvtl :Il'li-l' [ruling periods of 2 months to 2 

'l'ln: ninjnr clinic-:1! signs, in all groups. 
and :1 reduced weight gain. clininnl :nnl 
observations of [he L'xpm'imcnl?l animals will he rcpm'lctl else-
Erna lillili). Table 8 lln?: lissuc
levels of 2.4-1) found after a fc'ccling pc-riml of at lc'usl 2 months.

Tu li 9. Tissue low-ls ul' in pigs showing signs of poisoning
on l'OIH'?iL?ti Ul'ui ol? 2,4-1) amine.

(fresh Weight)
'l'issuc 

 

 

 

 

Pig no. 3 A Pi},Plasma 300 530' 530
Liver 230 300 225
Kidney - 260 370 185
Lung 22? 340 190
Spit-en 130 225 150
Skult'lal muscle 10:3 1'20
lirnin 40 (35
Urin IUU 90 

10 daily doses (if 50 Jug/Lg. liillcd 43 lmurs :Il'lcr 1215i. ?loamrepealed oral (if 2.4-1) :nninc
(50 and 100 ing were gin-n by in
5 and 2 pigs, 'l?wn animals of firs! and (mo of lilL'
SPl?JllHi gran]: (lowlulsml signs of poisnning (vomiting, 
and were killed. On analysis (if lissm-s
{rum lln-sc animals, lln- givun in 'l?nlalu El {Vt-re nlilninwl.

Furlln'r, {our pigs gin-n nrul ?luau-s nl? 
exit-r (?ll mg fur up In our inunlli. ln lln'w nl'
llu'rn' :inimnls m: i-l'l'ri-ls \wn- mud in lln-il' 
unl?x' luv.? lm'rl.? nl' \i'cn- l'unml ill). 
('ullili nul ilt?Ilii'll?l'il'li in lirmm?s l'l'mn :mimnlr?l. (3m-
:mim::l. lmn'rwr. signs ul' Jmiwning; :nnl mu. 

ili'lc?l' thaw (pig; in?. if? 'l'ul-h- 11). (In xix 

3258838 M00

16713

1]

Tu i 'l'isxue it'\'t'ls nt' in pigs after repeated ural admini-
stratiua ut' truly! esh'r (equivalent tn lug: l?nr
I Inunth. The animals were Lilled hnurs after' last duse. liaeh value
given is the mean nf results. {rum 3 animals, with the range gin-u in

 

 

hraelmts.

Tissue 2.14), (mfg; (fresh weight)
I'lasma 7 5 ?1.2 
Liver It) (5 --I7 
Kidney 17 (I2 
Lung 4 2 4- 7 
Skeletal musele. 1.5 0.5 3 
Fat 0.5( 0.2-- 1.1)

 

hiin tissue levels of 2,-i-D were found (230, 190, 205 and 80 ?14/3
in tin-r, kidney. lung and skeletal muscle, respectively. 

Finally. with the. main of studying the. pnssihle effects
of pheiny acids on relirmluctinn,-a pregnant sow Was given
2.4-1) amine (500 p.1un.) in the feed during the. entire pregnancy
(see lijc'irklunu' Erna). or fifteen piglets (Iclix'ered un parturition
one. was horn dead and nine died within 24 litiurs. On analysis
of tissues from the dead piglets considerahle. amounts at 
were. found; for liver, kidneys and lungs the mean values, and
the range. were 35 27 and 110 (13?6-1) g,
respectively. The. placenta contained ~16 of 2.4-1) and the
plasma of the sow at parturitinn 28 [cg/ml.

DISCUSSION .

Absarp/iun. when :ullninislered orally as a technical
f?rmnhitinn of an amine salt, was readilgr in all the
animal species studied, as indicated hy the peak plasma enneen-
tratinns after a single dose heing attained in a few hours (ahnnt
2 hours for chickens and flu-7 hours for rats, pigs and calves,
l-?igs. i?u?It). Similar results were nhtained with an alkali salt of
pure thus indieating the rate of to he
essentially unaffected by the amine ur ether ingredient (If the
technical furmnlatinn. 2,4,5-1? amine heha'Ved similarly.

Oral :nllninistratinn nl? ester, nu the ntln-r hand, resulted
ill hm" plasma and tissue levels of Bel-l) as emnpari-rl with the
snluhle salts. Ju'nhalnly as. the result nt? ahsm'ptinn
(Fig.5. Table (Innaith-I'ing; llH' luun' in (If

the :?slel', a hm aliunptinn rate is alr-n tn he eslu'rleil. Nut at 

16714

til

time after dosing; eoutd intaet ester he tll'ltit'H'tl in plasma or urine
of rats, pigs and ealves. (July the aeid \vas (leleetahle. indieatiugj
the ester to undergo hydrolysis during ahsorption (Ifrne 

Distritnttinn. 'l'he. ahsurheil pheuusy aeids \vere distrihutetl
rapidly throughout the laud); ('l'ahtes in all speeies? the
highest tissue levels were found in the excretory organs, liver and
kidneys. amt in the lungs and spleen. The levels in liver, kidney
and lung at (3 hours approached, and at later times often 
the plasma level. This was particularly true. for rats and hens.
A emnparison hetxveen species may he based on the. kidney levels.
If expressed relative to the liver levels, these values at (5 hours
and 2-1 hours, respectively, were 1.6 and 1.3 for pigs, 2.3 and 5.4
for male rats, 4.1 and 5.0 for female rats and 1.5 and 3.2 for
ehiclu-ns. 'l'tius. rats eshihited the highest values ttn'oughout, and
in hoth rats and chickens the values inet'eaSed (luring;th elimi-
nation phase. The difference in (3-hour values for male. and female
rats prohahty is not significant. Likewise, there seems to be no
sus-dift'erem-e in distrihution pattern in rats. tissue
levels found in the females consistently \vcre. inferior to those
found in the males.

From the more detailed study performed with pigs (Table (3)
is seen that compm'ativety high levels am he attained also in
endoerine. and other secretory organs. (Ioneentrations of 
comparahte to that in liver, or 50 5-2. of the. plasma level,
were found in ovaries and adrenals. In skin and heart musele
the levels were approximately and in skeletal mnsele about
10 relative to plasma. .

't?he hraiu level was usually low, ahont 5 21- relative to plasma.
although after losie doses higher eoneentratious could he attained
(see Table St). to adipose tissue. only ll'aees of 2.44) were found,
or 1?2 relative to plasma. The. low hraln level is ennsistenl
with eurrent eoneepts of drug transfer aeruss hotly membranes.
For most drug's amt foreign organie compounds the transfer can
he explained in terms of a simple diffusion. of the unioni'Aed
eompound, aernss a lipoid memhrane, the rate-det-ermining fae-
tors luring the ionization of the and the tipid-sotuhitit)?
of the form (see for instant-e llr'nns tt'til). phen-
oxyart'lie :Ieitls. as living relatively strung; neitls values alanut
ill. will he hugely l?tuized llu-i'et'me uI-t readily tlill'usihle at
plasma pll. they \vuuhl ttHi lu- esperletl lv 

llu' lalmnl-luaiu llalft'h'l'. as Well as the Illar'rulid Ivatlilv.

b38893 MOO

16715

. 

[his llu- wsulis uhluim-J will: [he 1111;411:1111
:1 rmdy jilzu'vulul Irunxl'vr uf 51-1-11! inlvr-
paling. 'I?lu- ?mulls :1lh'ihul:1hl1- In :lilTvn-nl l'urlurn, surh
:15 1'h;111;;r 11f plum-111.1! pvrunwlinilii?' (luring; [111-5111111133 luwvriug
nf plurvnlul pH :151-11111pu1'1-d I11 :1 1'111151-11111'1? hm?rr-
in; HF H11.- of iunixuliun 11f H11: phmun'y urhl. :uul uxlru-
plum-111:1] Huwvwr, Hu- 111'1'31'11l limilml (luvs
nul prrmii :111 :1551'551111-11! ml" [111: whllin- ilulun'lulu-r 11f 1h:- 
fut-101's.

'l'hu disll'ihuliun pullm'n of 1-3-1). :111111iuisl1-1'1-cl :13 [111' cslur,
Hal (liffvr :111111'1'1-inhl3' 1?1'11111 Hm! ?ht-owed \1'ilh amino,
:tllhuugh lissuc ('Ullhihh'li? 

'l?hv :11111111'1'111 distrilmlinu of [111' culupunnds under
study (1111 hy pinning llu- curve-s
(Figs. 1?4) 1111 :1 M'uh- (lug. 1'11111'. 111110.) and
cxlrupululing Ionninul. anm' l-nrt 11f Hm. plul hunk lo zero
lime. My relating the dose gin-u ln Hu- "zoru [itm- pluxnm [cu-ls"
thus uhluinud. apparent. (lislril-uliuu \?uhnucs nf uf
?11- body ?right were found fur H.1- sulls of, 2.4-1) :u1d 
when :Idminislcrod urully :15 :1 singlv dusc. These Values 
H111 oxll'm-clhiLw hudy vulumo (almul 2U 52-
nf hotly and [(11:11 hudy \mlcr (uhcml 7 ?23 of H10
hmly ?vighl'J. H1115. .xmuc [ac-1114111111111 (1f the phvuuxy
{wills in!? ?lo Dircrt fnl' Hu- of Mom}
lu ?2-15 uhluiuul in [he 511111-
in 3 and 1.111 H11: hluml veils I'c'gulurly ('uuhl
ho in ranging If) and '30 01'
[he (-111'1'1'51111111Ii11g plasma Murvuvci'. 111122.44) 1'111111-111
01' [111- (?uuhl hu 0:15in ?115111113, which
indimlvs :1 road); pussugn 11f 1111011115.)" acids :11?1'115?5 [he 
Thu distrihuliun cquilihrium hulwm-n plasma and Mom!
is uhviuusly displuwd inwards :1 furl whirh is Ms?
l't?rh't'lt'tl in plumuu 111-1111; surpvriur 1115;111:1111? wushiug efficiency.
0111- 112151111 for lwing ruuhl 
:1 puriiul him]ng ur lulu-Hwy :u'ids In lulusum lu'ulvins.
H1115 rmllu'ing ?11- frat-Hun Eal? phennxy?urids in
huhruliuus u" xurh :1 lu?uh?in hindini; 11:11'1' ulnluim-tl
(Ifl'nl' -

liliminuliun. Iilimiuulinu (If ?11? :11'i1fh hum plasma.
:1fl1-1' loving; :11l111311i-?l1-11-1l :1111'1111- 1.1 Imu. lull-M in

Hum-x.

aessamoq

41.6718:



were linear in their terminal eonrses. indiealing a first order
elimination rate. The plasma half-life values ralenlaled l'i'ozn
lhese earn-s were almnl hours For l'als, 5 hours for ealres and
ehiela'ns and 12 hours for pigs ('l?ahle. No sex different-e. in
respeel to elimination rate was ohserved in rats, although the
plasma levels were lower in the females than in the. males.

The elimination rate from plasma of given as the ester.
was redneed as compared with the soluble salts.

The rate of eliminaliim of 2.4-1) from tissues was lower lhan
that from plasma ('l?ahles From the. lahnl'aled dala, hall'-
lil'e. values of ranging between 5 and ill ho'nrs can he eal-
eulaled for rats and between ill and 30 hours for the other speeies.
Generally, in 72 hours or less, tissue. lerels had dropped to a
few [lg/g. retention of pin-nos)- acid was noted in the tissues
examined. The results are in agreena-nt with those olilained in
sheep using,r (Clark at 196-1). The distribution
pattern of on repeated administration was essentially
similar to that obtained after a single dose. although individual
variations were considerahle, presumably owing to variations in
feedng hahiL of lhe animals :md in lime. hct?w'een last waler or
food intake and time of killing (Tahles 8 and 10). The resulls
seem to support the View lhat halogenated phonosyaoelic acids
are not likely to aeenmnlate. in .lissnes of clinically healthy
animals.

Erma/ion. Judging from the. analylieal results the major
excretory mate is via the. kidneys, at least in the. mammalian
species studied. Ilislopathologieal examination suggests that this
is true for as well (lijiirlcluml 1960). Only low
levels of were. found in feces from rats and pigs and in bile
from pigs. 'l?hal phenoxyaeelie acids are. predominmitly excreted
renally in cattle. and sheep, was reported by el al. (1963),
Buelu' e! (lilti-l SI. John (il. (Will) and Cloris e! 

Interestingly, hens were found to he. lo eserele part of
ingested 2.1-1) with the eggs ('l'ahle a filnling'nnt previously
reported. Mosl. ol' tln- eonlent hi the. egg was found in lhe
yolk.

(ln prolonged of 2,-l-ll. given as
amine or as esler in elm-u". ln-h-w lln- losit' level. Mime
espriinienlal of an adaplalimi. ln pi;;x.

plasma lu-n'l'. ul' slal'lell In liq'L'iliH' allm' aim-.11 a

039823 moo

16%17

15

\vm-lc. lln- nrinnr)? (Table 
nu. -l and 11A). A similnr \vnx' arm in rhirin-ns.
ll Ju- hmx?vrur, pigs 501:?! In In: 1cm;
init'rnnl In L-le-snru In Iiilt'lH-xb' ln-rliicidc?s lhun are
1111- an-r spvc-it-x slndirci. In instant-rs, given :15
(1:1in (lnws nf 5? nIg/lgg, tiny ln'mhn-od signs nf' puisun-
ing; in pig's: will: rising;r and lissnc low-Is nl' 2.4-1) 

luml lirnr).



Audus. L. 1.: The I?llysiulngy and u! Hcrhicidvs. Acu-
dt-lnic l?rrss, London .Q New Yuri; 100-1.

Jim-hr, (I. 21., I). J. List, I). G. J: G. Wm'm'r.? I-Ililninnlinn of
acid and 
plu-nuxgr)innyric acid in the urine. from cows. J. Dairy Sci.
1110-! u, 47, 93?95. .

Ullt?ht', C. 11., I). D. Haru'cc, R. lluilnml J: I). J. Li'sk: Absence of
ln-rhiciJc residues in the milk of dairy cows at
high frvding levels. J. Dairy Sci. 190-! 

Binnx, IL, ML: .?thsorplinn and Distribution of Drugs. 
Edinburgh .31" London 106-1.

(C If. Erna: studios: of 
herbicides. Acln rel. scand. 1066, 7. 1n prL-ss. 

Clark, I). J. I'unng, R. L. L. M. Hunt J. IL'JIc?fmruu:
'J'hc fate uf acid in shot-p. J. Agric.
Fund Chem. 1901, 12?, 43?--15.

Erm', AX: Holt-rminnlion u! pin-nuxynut-lic herbicide in hi0-
lngicnl Innlorinis. Acin M'nml. 1906 u, 7, 775?410.

If; Slndirs on animal uf horlai-
CillL'S. Arm 19061;. 7. In press.

I). I). Hurdrc, R. (E I). J. List?; Dis-
of :lricl hvrhicidc in
line dairy J. Dairy Sci. 1963 ?16, 

Gulcunmnn, W. 11., I). llut'gc'r. If. llulinml D. J. Lislc: Residue

_.~.Intlirs will: :zc'itl hrrhiridv in llu-
?luiry nm? and in :1 natural :nnl :n'lifiriul rnanun. J. Dairy Sci.
?Hi3 h. 1257 1283.

lat'l'f?fl- H. H. 'l'lu- nu-inlmiimn uf acid, ilx
and .wnn- iu'itih. 
Hi B-midnlinn. J. (:llt'Eli. 213 2313!.

LIX-J7. I). H. Bur/Ir, H. H'iu'tu'r' H. (I. 
m-r: l'ZHHIinniinn ul' I1 in [in- nrin. u! 'nil'l'l?\ it'd 
m' 13;! H, J. I'Jiih. if}. 

{mesa/woo

1:6 

1r. 

St. Julm. l? 15.. I). ll'uym-r I). J. la'sl': Fate uf alrazine, Inn-nu,
silrex and in dairy J. Dairy Sei. l. -
1270.

ll'illimus, It'.  Ilelusiealirnl Meehanislnx, lid, Chapman llall,
anlmt i939, p. 33?.


'l'he dislrihutiun and elilninatinn at' twu phennsyaeetie aeitls,
2,-1-11 and Were studied with a ehemieal inethml in rats, pigs,
:alres and ehiekens.
\\'hen :ulmiuistered orally as amine tn" alkali salts, the eompuunds
were readily almurhetl and distributed urer the nrganism in all species

studied. The of 2.4-1) in the form at? an ester was incomplete. .

huwerer, the ensuing,' plasma and tissue levels of being only haw.
ester Cnuld nut he detected in plasma).

The highest tissue. levels- at 2,t-l) and Were found in liver.
kidney, lung?r and spleen, the levels sometimes esteedilli,r ?It? lilil?lH-?l
level. In eells 10?30 ut? the plasma level was found. l'ene-
tratiuu u! 2,-t-D iutn adipose lissuu and into the central nervous system
was restricted, whereas a ready placental transfer was demonstrated
in swine. The dislrilmtiun pattern did nut show any signifieant species
m- in rats sex ditferenees.

liliminatiun ul the emupuunds was' rapid, the plasma half-life
being about 3 hours in rats, almut 8 hours in calves and Chickens and
almul 12 hours in pigs. The. tissue half-life. values ranged between 5
and 30 lunn's, the lower \?aths heng iuund in rats. Nu retentiuu in
tissues was nuled, nor was :uzeumulatiun seen repeated admini-
slraliun. 

ln pigs and ehieltens an inereased eliminatinn rule was nip-erred
atler repeated :ulministratinn. -

The Major exert-they route seemed to be via the kidneys in all



speeies studied. llens excreted small amounts u. with the eggs.


l'erleilnng nan lilimiimlimi mm ehlurz'erlen I'm


Die Verteihmg und lilnninatiun \?un ehlurierten l?ln-nnxy-
essigsiiuren. uud wurdenen ?atten, Selnveinen, 
lliihnern untersneht. 

Naeh nraler in Farm der mier Alkalisalxe. wurden
die Verhindungz-n ln-i allen nntersm'hteu 'l'iez'arten sehnell resnrhiert,
Ilml iihel' den Organisnnm 

(ll'ale Zuiuhr run in l"urm elm-s ereah jedmh nur
niedl'ige l'lasma- und run Ari-ll, anwlu-inend lul'nlgg'


Naeh ehwim-r ?rah-r Kat'nln \'t ll 
\x?nrdt-n die him-hate? hivrwilen den l'lasnmv

spie;;n-l in la-hcr. Nitre, \liL'. {timid-us. ln

gaessamoo



l7

ill'll run-n ?in-20 ?It-s nm-h-
2,14) nur in nimh'im-r Kunm-nll'nliun ins 
ins. Hum-gm: wurtlv (-iu hurclulrin-
urn dc-I' I'lm'L-ulzuimrricrv lu-im Schwuin In: 
Iwiuc (In-i Hulk-n) und nur Iniih-
hiifl' Ynl'iuliuncll 

DEL- .wlum-ll dic Hulbwt-rlzt-il in
Plasma. Much cinzigcr nrulvr liingnlu'. hch?ng Cu. 3 Slumk-n hci 
ca. 3 Slumlcn hci [{Eillml'n Hi'lhm-rn uml (3.1.12 hL-i
Div lInltm't-rlzt-ilrn in (It-n ('wwciu-n \vnron lmi Hulk-n
5mm Slmulcu und 110i Kaine
Vun c-lmnso kcinu
At-Iaunmluliun Zufnln'. ..

?wi 11nd inihum'n in cinigrn I?Eillrn nurh
an?ulu' cine genlcigt-rlc I?hcnuxysiinrml
7

Die \x'ln'dcn hei nnicrmu?htcn Arlen han'pl-
.siivhlich (hm-l; div Nicn-n ansgt-Mthic-dvn. Evi Hunncn wurdc cine
vun Mcincn Mongcn 2.4-1) mit Iiivrn nachngicscn.


hislr?ruliuu clinu'nm'ing an [clot-crude 110:: djm'.

och dislrihnlion och
clinlincring grimly knlvur och 116m; 

Tillf?rdn m'nll mnin- rf'n'vnin-
Iiill (ICII {GI-(Iclzulcs snuhhl hm; nllu undurs?ildn


liIIc-r [ini'n'scl ('hlt'l' 151g? 
lwrm'ndu pa?l nfullsli?mlig rvsurpliun.

va-r, njnru, uch ijillu Vismic dc hi'rgslu 
\?illia iblunul (Hersh-g film-[anus ?j'o
:w ut'h crnh'uln on?
i ring? grad 3.14), Ii?. sym-s uhchin-
hm; .win. Dialrilmtinnsmiinsh?c?t visimlu cj might signi-
fikanf fiir I'?llor. Hmn'?lhu' \?iszulc dock n?gul
153;;1'1- m'h 

lmh?cringslith'n 1' plasma var
3 bin!? kiimu-n, can 8 linh'nr m'll
hiins 11?. fi'n' gl?IsuI'. Huh't-ringxlitlcn i vur
limmur; I'?Hm?nn luulc do liigrv \??l'dcnn.
rclu-nlinn i t'j lwlh'r "3.361: :u'lul-
mulnliun "ppm-put! lillf?rsci.

Hus grisnl' nlc'h Ili'mx nlxi'antll'iug (?flrr
ltplu'rpml :nv 2,1-11.

l'lxa'i?ch'iH?vH g;mm:n njururnn haw. ?mirr-
M'ukln ark-r. Haw. Iniin;;IIrr ELI-H 

(NH rl'r'u'rf Hurt/1 lf?lili).

men/woo

ca

to
Eu).
:3

D. Toxic Effects of Herbicides and Conclusions

 

As mentioned in the introduction of this chapter, we are listing
here a variety of information which could not properly be listed in the other
sections. The items specifically included here are the following: (1) the
mutagenic activity of herbicides on plants, bacteriophage, and bacteria, (2)
the effect of herbicides on the embryonic development of beetle, (3) the con-
centration of herbicides in the tissues of fish, (4) the importance of select-
ing a formulation which will minimize the_hazards of herbicides in aquatic
environments, (5) an estimation of the hazard involved in the spraying of a
man with herbicides,and (6) some general conclusions about the toxicity hazards
relative to the use of herbicides. 

Mutagenic Effects

 

Some of the persons who are critical of the use of herbicides feel
that we are creating a monster which will eventually destroy us. One area of
expressed concern deals with the possibility that herbicides will produce
heritable changes in the genetic material of plants. The resulting plants, they
fear, will no longer meet our need for the production of foods or they may be
insidious weeds which do not respond to either chemical or biological control :3
measures. Some studies have been conducted to determine the likelihood that C)
herbicides may produce heritable changes in plants. is

Doxey and Rhodes (1949) conducted a study of a plantugrowth-regulator 
?type of herbicide known as acid (morn). In


this study they exposed onion root tips to MCPA and observed the effects of pg
this treatment on cell division and on the chromosomes. Chromosome 
and breakage were observed and the effects were judged to' be similar in many 

respects to changes in mitosis produced by X-irradiation. In a series of three ra_
papers, Unrau and Larter (1952) and Unrau (1953 and 1954) discuss their findings 
on the effect of on the genetics of wheat and barley. The seed grain

fields were sprayed with 8 to 16 oz of (EE) per acre 30 days prior to
cytological examination. The chromosomes were studied in detail in those_

cells that were dividing during the process of pollen formation. Regardless

of the date of application or the concentration of treatment,the herbicide

caused ?highly significant? abnormalities of chromosome behavior and the

changes were visible as much as 50 days after treatment. Unrau warns that
indiscriminate use of herbicides could have grave and far-reaching conseQuenCesj.
especially in relation to the prime seed stocks of seed growers. There is

reason to suspect that the herbicide interferes with self-pollination and this
could explain the increase in the number of off-types or crosses obtained in

Unrau's experiments. 

201 167-21

DOW120472.

Muhling et al. (1960) has studied the cytological effects of 2,4?
dichlorophenol, 2,4-dichlorophenoxyethanol, and other herbicides on
pea seedlings (Pisum sativum). was clearly shown to be a prophase
poison and to produce spindle inhibition resulting in chromosome configura-
tions very similar to those produced by colchicine.

Dr. Kenneth J. Andersen (1967), Senior Research Microbiologist at
Battelle Manorial Institute, Columbus Laboratories, is engaged in a research
program for the United States Department of Agriculture to determine the pos-
sible mutagenic and genetic effects of the herbicides. As a part of this
study, he grew histidine-requiring mutants of Salmonella typhimurium on a
histidinesdeficient medium to screen 120 of the 130 herbicides listed in the
journal, Weeds. These included, 2,4-13, picloram and cacodvlic acid
and the oth?r herbicides which are currently important for noncropland 
vegetation control (see Table the-s. typhimurium used is his-tidine-
dependent it does not grow very well on the deficient medium unless there is a
mutation which eliminates the histidine requirement. In his experiments he
found that diethyl sulfate, beta-propiolactone, acridine mustard (ICE-91) and
all produced the mutations required for
the growth of the organism. On the other hand, none of the herbicides caused
this mutation.

A second approach to the problem involved the use of T4 bacteriophage
to detect chemically induced mutations of the type. The number of mutant
plaques, which characteristically were larger, with a clear center and a sharp
edge, were compared to the normal T4 plaques which were small, with a clear
center surrounded by a halo. In addition to this characteristic plaque
morPhology, mutants form plaques on Escherichia coli but not?E. coli KB
while wild type T4 bacteriophage forms plaques on both of the E. coli strains.
In,this evaluation system noninhibiting levels of the compounds (approximately
20 us-l mg/EO ml) were employed. The rate of spontaneous mutation (no test
chemical present) by T4 bacteriophage to the rII-type mutant, was about 0.11%.
In the presence of the known mutagen, 5-br0mouracil, the mutation frequency
increased to about In the test system it was concluded that none of the
herbicides were mutagenic.

Dr. Andersen has also performed experiments using a 2-aminopurine-
induced mutant of T4 bacteriophage to detect revisions from the mutant to
the wild type which might be caused by the herbicides. Herbicides, in this
detection, were again observed to be nonmutagenic.

McGahen and Hoffman (1966) have also investigated the possible
mutagenic activity of herbicides. In their program they employed a screening
technique similar to that used by Andersen (described above). In their study:

is 57.22,



they used both the 3- and also the 6-alkyl bromouracil herbicides (including
bromacil) on a bacteriophage and concluded that it was not mutagenic by this
test.

Dr. Verne Comstock (1967), U. S. Department of Agriculture and
Agronomy Department at the University of Minnesota,has conducted a seven?year
study on the effect of herbicide applications on the growth, yield and quality
of flaxseed. During the years of 1956 to 1965 paired lines of four varieties
of flaxseed were grown on plots which had received no herbicide or had re-
ceived 15 to 20 oz/acre of MCRA per year (treatment was omitted in 1959).
The performance of unsprayed subpopulations were compared with that of the sub~
populations which had been sprayed in the F2, F5, and F4 generations. "It 
can be concluded that the application of high rates of MCPA to successive
generations did not cause a significant shift in these flax populations to
more resistant biotypes.?

Insect Development

 

Several investigators have considered the possibility that pesticides
may under certain situations cause a disturbance in insect population which will
result in a stimulation of a secondary pest. The reason for the increase in
aphids in the grain fields in which amine has been used for weed control
has now been investigated by Adams (1960). In her study three species of
coccinellid beetles (Coccinella transversoguttata Fald., Hippodamia
tredecimpunctata (L.), and Coccinella perplexa Muls.) were grown in the
laboratory and Sprayed with at various stages of their larval develop-
ment (1 to 12 days). It was observed that spraying on the 12th day of larval
development produced the highest mortality and about a 75% increase in
time to maturity. Since this bettle is an important factor in the control of
aphid populations on grain, weed control with herbicides should be timed so
as to minimize the damage to the useful beetle.

 

MOO

 

61.17031

Concentration in Food Chains

One of the his concerns of the ecologists is this,are the herbicide;
concentrated at some point in the food chain so that some life fonn at or
near the top of the food pyramid will receive a toxic dose? Fortunately there
are few life forms which concentrate herbicides at all and in most species
this concentration is a reversible process so that the concentration level 
remains small. Table shows the amount of herbicide present in the tissueS
of fish after exposure to the concentrations shown. For comparison it may be
noted that under these same conditions heptachlor accumulates in the bluegill

203

"Jus723



V08



Species
Bluegill 
Redear Sunfish
Rainbow Trout
Green Sunfish
Bluegill
Channel Catfish
Bluegill

Bluegill

 

Fenoprop.

Source: Cope, 0.B., Contamination of the Freshwater Ecosystem by Pesticides, J.
35-44 (1866). 



TABLE 

AMOUNTS OF SOME HERBICIDES MEASURED AS RESIDUES

ANIMALS AFTER 30-DAY EXPOSURES IN PONDS

Pesticide

PGBE in oil

Silvex,* PGEE in oil

Paraquat emulsion
Paraquat emulsion
Paraquat emulsion
Paraquat emulsion
Sodium arsenite

Diquat

Locality
Tishomingo, Okla.
Tishomingo, Okla.
Denver, Colo.
Denver, Colo.
Denver, Colo.
Denver, Colo.

LaCrosse, Wis.

LaCrosse, Wis.

Amount of
Pesticide in

Water i232!

10

0.23

Amount of
Residue in

M)
None detected
0.5
0.11
0.05
1.21
0.57
0.40

0.09

Appl. Ecology, (Suppl.)

to the point where the tissue level of heptachlor is 300 times as great as the
heptachlor level in water in which the fish is swimming (Cope, 1966),

Selection of Herbicides

 

At the present time there is no magic formula which will automatically
select the herbicide which will control the vegetation best and provide the
minimum damage to fish and wildlife. Instead herbicides must be carefully
selected on the basis of the weed that needs to be controlled, the location
of that weed, and the possible fish and wildlife which may be affected by
its use. That selection of the proper formulation for the job is exceedingly 
important in the control of aquatic weeds without unnecessary toxic effects on
the fresh-water ecosystem. In Table VI-ll, the acute toxicities of a series
of herbicide formulations to lake Emerald Shiners (Notropis atherinoides) are
presented for three different exposure periods. The importance of the formula-
tion employed is borne out by the observation that both the most toxic and
the least toxic formulations are formulations of endothall and one is about
6,000 times as toxic as the other.

Spraying of Herbicides on Man

From an examination of unpublished results from the Patuxent'Fish
and Wildlife Center at Laurel, Maryland (Robert G. Heath, 1967), we conclude
that dieldrin is at least 30 times more toxic than amitrole, atrazine,
dalapon, diquat, diuron, monuron, picloram,
simazine, and acid to the two-week old young of mallards,
ring-necked pheasants, bobwhite and coturnix.

One further comparison of the toxicity of herbicides and pesticides:
the Federal Aviation Agency is much concerned about the toxicity of pesticides
and their effect on the ability of the pilot to fly his plane safely.

Dr. Paul Smith, Head of the Pharmacology Section at the Aeromedical Laboratories
in Oklahoma City, said that the pilots spray insecticides during the early part
of the year and then spray defoliants and herbicides in the summer for brush
He said that the pilots themselves are always glad when they are
through spraying the insecticides. He added that the flagman on the ground
during the aerial spraying operation probably receives a higher dose of
herbicides than the pilots. So far as he is aware there has not been a serious
case of poisoning or toxicity as a result of the spraying of herbicides.

 

BEE Dutoxyethyl ester; DMS Dimethylamine salt.

205

MOO



16725

DOW120476

TABLE 'V?I-ll

CCMPARISON OF THE ACUTE TOXICITIES OF COMMERCIAL HERBICIDES

 

Product

Aquathol-
Silvex

Weed

Rhap 20
Amitrol 

Kurosal 
Crop Rider
Kurosal SL
Reglone

(DB)
Reglone

(DC)
Atlas A
Esteron

99
Garlon
Urab

Kuron
Hydrothol

Source:

TO LAKE EMERALD SHINERS (NOTROPIS ATHERINOIDES) IN

MEDIUM HARD WATER. MEAN TEMPERATURE 69? - 725F

 

Mfr.
Pennsalt
Reasor-

Hill
gmchem

Dow



Dow
Chipman

Chipman

Chipman

Dow

Dow

Allied

Dow
Pennsalt

Soc.

 

amine.sa1t

206

TL

 

 

 

Liq. Active mliPPm aetive
Gr. Ingredient Endothall 1,000 780 612 510
Silvex
Gr. ethyl- 1,000 620 620 510
hexyl ester 
L. Aminotriazole 910 455 455 420
am. thio- 
cyanate 
Gr. Silvex, potas- 1,100 540 450 370
sium salt
Gr. ethyl- 500 280 280 280
hexyl ester 
L. Silvex, potas- 509 420 510 270
sium salt
L. Diquat, di- 180 180 86.2 25.8
bromide form
L. Diquat, di? 180 15.5 11.7 9.1
chloride
form
L. Sodium 32 15.5 8.1 8.1
arsenite (As)
Gr. 2,4,s-T 10 4.5 4.5 4.5
butyl esters
L. Dalapon 10 4.2 4.2 4.2
Silvex (butyl)
L. 3-pheny1-1, 5.6 4.7 4.3 .5.8
1-dimethylurea
trichloroacetate I -
. L. Silvex, butyl ester 10 4.0 2.4 2.0.
L. Endothal1, coco- 0.29 0.12 0.10 0.08

Y.H. Swabey and C.F. Schenk, Proc. Third Annual Aquatic Weed Control
1963, pp. 20~28.



?16725?

. ?arW?ab Ale i eSbl mes Domg Human Bangs? - 
 I - 
REVISED and UPDATED 
 '7  TWENTY YEARS. 7 .
- i: GRANVILLE  KNIGHT, IM.-D., F.A.C.A. 
- .- IPreSidcntPRICE-POTTENGER NUTRITION FOUNDATION. 

 

 

 

 

I I. j_Publislmd In MODERN NUTRITION MAGAZINE  April. September 1952Copyright 1972 by Author nup- 





.



What Are Pesticides Doing to Human Beings? - fr
- By Granville F. Knight, M.D.. F.A.C.A. .
Santa Monica. California 90403 - - .

the publicity agents of new poisons added to the diet. This is

it?-

'1

 agree.
minds of those who-have personally ex-

of the insecticide 
terieal alarmists" is the quaint descrip-
tion applied to some of those who even
suggest that the public is being harmed
by their use. Groups urging that the
newer pesticides not be released corn-
mercially until proven harmless to hu-
mans by exhaustive testing. have been
accused of obstructing research and scien-
tific progress!

 

?Nothing to worry most
of our entomologists. ?Pesticides are poi-
sonous. Yes. But with proper precautions
those handling them are safe. And with
the establishment of tolerances for resi-
dues on food the public is protected.
Moreover. farmers can not to bar-
vest the large crops needed throughout
the world without the newer pesticides.
Better the risk of same toxic residues on
our food than widespread crop failure
from insect attack." These conflicting
statements remind one of the cheery
whistle adopted by a small boy when

passing a cemetery at night.

"Danger!" Widespread illness from the
Use of DDT on craps and in homes. This
is the belief of a few keen observers in
the medical profession. A handful of agri-
culturalists. entoniologists and
There is no question at all in the

perieneed DDT poisoning,

How may we account for such marked 
it 15 not difficult. -

divergence of opinion?
The manufacturing companies are con-
cerned with a booming business. The
entomologists' responsibility lies mainly
in protecting our crops frOm insect pests.
Both groups are loath to accept evidence
which might interfere with the product"
tion and use of the new insecticides.
Knowing: that some individuals exposed
to large amounts of DDT remain in ap-
parent unod health, they refuse to believe

- that small. or even lame. residues on food

might be harmful to the general public.

Physicians. on the other hand. are pri-
marily concerned with the health of the
people and are quick to sense the threat

Reprint?! from The (Vader:- anririon {Vain-inc --



others .

particularly true in view of the fact that
DDT is very stable and may be concen-
trated in body fat up to at least 30 times
the level in the food consumed. Unfor-
tunately, scientific proof of toxicity to
1hurvians from low levels of intake is dif-
icu t.

The potential hazard to health is tre-'
mentions.

It must recognized by all
parties concerned and controlled until
the facts are established beyond all deuth

The Delaney .Committeo

In.spite of the assurance that there is
nothing to fear. the_public has become
alarmed over the widespread use of pot-

sonous sprays and dusts on our crops. 

And so.

This alarm was communicated to the
Congress which appointed a Select Com-
mittee to Investigate the Use of Chemi-
cals in Food Products. The chairman is
John J. Delaney. M.C. In addition to
pesticides. this Committee is investigating
the use of chemicals in other fields. such

as drugs. cosmetics. food preservatives. 

and fertilizers. 

For the past two years the Committee
has been hearing testimony from promi-
nent scientists.- entoinoineists,
ists. load growers. manufacturers, physi-
cians and others interested in the prob-
lem.- - 

in the field of pesticides. testimony was
overwhelmingly against the idea that the
public health was being endangered by
their use- Hewever, a few physicians pre-
sented convincing clinical and laboratory
evidence of widespread illness from con-
tact with DDT on leads or in sprays.

Louis Bromfielcl was not only eon-

vinced of the hazard but from his own 

experience told the Committee, that with

correct farming methods. few insecticides  

were necessary or desirable._

Recently C. Cox. director of tech-
nical projects for the Beech-Nut packing

company stated that his'firm had spent 

$668,000 in the past six years in an at-
tempt to keep the newer pesticides out
or' baby foods and peanut butter. He also
stated that a fitntLuf $ll?l,00fl had been
mixed by the manufacturing chemists "to
counteract unfavorable publicity." Mr.
Cox related that in a number of instances
his company-had been forced to reject
large orders of fruits and vegetables be-
cause of contamination with DDT. Chlor-
dane and Benzene llcxachloride.

In a preliminary report_ to Congress in 
January. mat. the Committee recognized 

April, and Drums", i-r 

nutrition- 



?1 Tythe existence of a definite hazard to pub-
lic health and continued hearings. These
are still taking place. As a result of the
evidence it is to be hoped that remedial
legislation will be recommended to the
Congress.

The purpose of this article is to give?

the members of the Academy the latest
information on_tliis highly controversial
'and extremely important subject.

Extent of the-Problem

For the past thirty years or more there
has been a gradual increase in the amount
of poisonous chemicals used on crops for
the control of insects and fungous dis-
eases. During the same interval a corre-
sponding decrease in the protein content
_of our plants and the nitrogenous content
of our soil has been noted.

Since the introduction of DDT and
similar compounds in 1946 the tonnage
of the new insecticides in use has tre-
mendously increased. while that of the
elder chemicals has dropped. The marked
toxicity'of these new compounds for most
insects and animals. together with their
unusual stability, has .exchanged new
problems for old.

It is no exaggeration. to say that at the
present time. with the exception of fish.
it is a rate article of food that. has not
been dusted or sprayed with one or sew

eral poisons before reaching the con-.
sumer. Never before has the long-snitch-

ing public been made a guinea pig for
the mass testing of so many exceedingly
powerful chemicals. .

Food Contamination

Theoretically crops are treated long
.enough before harvest so that weathering

has reduced the residue below toxic levels. 

Practically this safeguard is of little com-
fort for the following reasons: 7 .

Enforcement of regulations is not

feasible except in the case of large farms;

It is physically and financially impossible

to police all the small farmers. llow often 

has ignorant enthusiasm led to the appli-
cation of many times the recommended
amounts? Who knows? And how many
small producers. if faced with the destruc-
tion of a crop by insect pests shortly be-
fore harvest. 'would have the fortitude to
refrain from spraying at that time? And
why should they hesitate? The trade jour-
nals tell them any danger to the consumer
'has been greatly exaggerated._l 
. (2) When airplanes are used for dustin
fields. drift to adjacent areas is frequent.

This has been reported as leading to inno- 

cent. btit heavy contamination of some
crops. . - -- 

(3) While our state and federal inspec-
tors are conscientious and hard-working.
they are-limited in number. One Wonders
how much food grossly contaminated .with
poisons reaches the consumer. We know

that same docs. Spot check tests can dis- .

cover only a portion of it.

(4) Our knowledge of the leng range
toxic effects of most of the new cem-
pounds is woefully inadequate. Therefore
how can tolerance limits be arbitrarily
set? Particularly when no tests have yet
been perfected for even detecting the
most recent ones. Logic dictates that until
all the facts are ltnown any residue on
food is too much. 

Other Contact: I.

As though poisonous residues on food
were not enough. the_public is exposed to
powerful chemicals in other ways.

For some unknown reason?and in spite 

of evidence to the contrary?we have been
sold the idea. that DDT and chlordane

are harmful for all practical purposes only-

to insects. The main basis for this impres-
sion seems to be these facts: (1) A com-
paratively few. healthy young men. when
exposed to' considerably larger doses of
DDT than would be encountered in or-
dinary usage. showed no ill effects. (2)
Animals exposed to DDT aerosols came
through unharmed except for local irri-
tation. No allowance has been made for
the fact that many humans are malnour-
ished. allergic and chronically ill; nor
for the fact that the young are particu-
larly susceptible to toxins of all kinds-

As a result of this propaganda. few)

households are without DDT aerosols or
sprays for fly control. And where anus.
are a problem chlordane (even more toxic
than DDT) is used for their destruction.

- Both of these substances may be encount-

ered in hotels antl restaurants. Naturally.

be feared. - - 

Most commercial tree sprayers use the
chlorinated hydrocarbons. in. addition to
this source of esposure. pleasure drivers

contamination of food and utensils is to_

on the highways in agricultural areas may 

be dusted by airplanes. I
- Is the Public Protected?

At the present time the Food and Drug --

Administration offers some protection.
but only has jurisdiction over interstate
commerce. The great weakness of the
agency is this?while it has authority to
set tolerances for pesticide residues odor
in plant and animal foods. under 

'law it can not prevent the use of a chem-.
ical before a tolerance has been set and; I
the relative safety of the chemical 

lei-mined. .
lost of the. state laws are similar.

Therefore- the state agencies labor under

similar handicaps. -
Potent new compounds are being syn-
thesized at 'an amazuig rate. Rough lethal

doses for animal life can be determined'

in a few months by acute toxicity studies
in the rat and larger animals. Llowever.
to learn the effects of repeated

?ll

at

?0

'mins undoubtedly remains.

contact with small amounts of a chemical
requires careful observation over several
generations. This may take two or three
years. in addition to being time-conSum-
ins. It is 

In the meantime the partially tested
pesdcide may be manufactured. adver-
tised. sold, and widely used! Is this pro-
tection? .

It is only fair to state that some manu-
facturers insist on thorough int-estieation
before releasing: a new product. Others
are not so careful. partly because they
lack the funds and faculties for such work.
and partly because the law does not de-
mand it.

Under the present laws any company
wishing to use a new chemical in or on
food is not required to first consult with
the Food and Drug Administration rela-
tive to merits or potential harmfutness.
This agency may confiscate or condemn
products only if they contain chemicals
proven to be deleterious. The inadequacy
of this law is obvious, but some illustra-
tions are illumination. .

Agene was used for artificially aging?
flour over a period of thirty years beinre

it was discovered that dons fed with bread
made from such flour den-elepcd "canine.
hysteria" or ?running tits." The use of
this substance has now been 
by the baking fraternity in favor of a
less 2' harmful substance. Dcnatura-
tion of- the proteins and damage to Vita.
Incidentally.
the United States Supreme Court in li'tlil
declared bleached flour to be
human_consumption. If this judgment is
still on the books, why has it never been
enforced? -

\Vlien the second of the "sulfa" drugs.
sulfanilarnido, was introduced some years

ago. in addition to tablets, a liquid
preparation sought. The drug was
relatively insoluble in the. ordinary sol-

vents. hut finally a chemical was found
that would dissolve an adequate dose.
This was one. of the group now used as
an anti-freeze in ear radiators. \Vitiinut
adequate animal tests the product was
marlictnd. It was condemned only after
a number of deaths due to __tlie solvent
occurred. 

Other examples could be cited. How
long must we wait before 
testing may be counted on to prevent
tragedies of this sort?

The Older Insecticides 

Let us now consider lltt.? more impor-
tant chemicals tisi-d for the destruction
of pests. First of all we shall discuss those
In use for many enough
to brim: tremendous crops to maturity
during the recent world 

With the exc'ption of selenium and
arsenic, these chemicals do not. transmi-
grate from the non-edible portions of a
a

unfit for 

plant to those parts used for food. Nor
do they penetrate to any extent into the
fruit or leaf. Therefore. most of the resi-
dues may be removed by scrubbing with
Soap and water and/or by dipping for
one minute in dilute hydrochlonc acid..
The chronic toxicity of arsenic is well
known. In small doses it may have a tonic
effect. "Arsenic eaters" in the 'l?yrol feed
it to their horses to increase their stamina
and eat it themselves for the same reason.
The custom is not highly recommended.
As the chemical is usually metabolized
and excreted in from one to six weeks.
high chronic doses are necessary to pro-
duce damage. . -
However, such high contamination may
occasionally be found. Livestock grazing
in orchards which have been heavily treat-
ed with arsenie compounds may-show
scriOus from such fodder. Some
years ago the American Medical Associa~
tion condemned apples from one county
in the State of Vashington. These 01'-
charcbt had been so heavily treated with
arsenic that dangerous amounts of this
chemiCal had been taken up by the roots
of the trees and translocated to the fruit.

Arsenic is often applied to fruits in the
form of lead arsenate. The limit for the
compound on apples was set in l9-ifl at
3.6 (parts per million).
component is more dangerous in small
amounts than the arsenic.

cause kidney damage. A high level of

calcium dict tends to be protective.

Mercury is highly toxic and residual
compounds are dangerous. As little as 0.5
may induce storage in the kidney
with resultant harm to that important
organ. Mercury vapor used to preserve

'grain in storage has not been shown to

contaminate such grain. 
Selenium in the soil is absorbed by the

roots and transniierates to all parts_of-
plants. The same process occurs following -
This substance 

the application of sprays.
may not therefore be used on food crops.
It causes abdominal pains. an ?l?l?llit. weieht
loss and liver damage. Cattle. Cropping
piants in seleniferous areas such as parLs
of Wyoming. develop the "blind staggers."
This is followed by paralysis and death.
Selenium sprays are used only on orna-
mental Ilowers. . . -

Fluorine compounds are less toxic than
lead arscnate. Their toxicity lies in their
aliilily to precipitate. calcium in the lmtiy.
?Clry'nlile? is used as an apple spray. \Vlien
so used the shipper is required to wash of?

the residue. Drinking water from I 

0.l to 1 has some effect in_lowerui::
the. incidence of dental caries In yountt
children. Even these may
ireduce mottled enamel and interfere with
)one metabolism.

The lead_

Lead is stored 
in the bones at all dietary levels and may 

I
. .


Bizarr-

 . 

v- 

'eides may be

Nicotine is one of our most powerful.
rapidly-acting poisons. One grain is the
minimum lethal dose for man and even
one-fifteenth of that may cause severe
It is a nerve poison. causing
stimulation which is rapidly followed by
depression. The alkaloid is volatile and
therefore presents no chronic toxicity-
Some of its salts may provide a residue
hazard.
no problem except to the applicator.

The pyrethrins. These were originally
obtained frein the petals of aamember of

For all practical purposes it is

the family grown in the,

Far East. analogues are now
available and have similar properties.
These compounds are rapidly detoxified
in animal tissues and thr'tethal dose is
very high. Therefore. they are compara-
tively safe. However, they may produce
severe asthma or other allergic reactions
in those sensitive to ragweed pollen. 
Roienone from derris root is most poi-
sonous to fish and not very toxic to man.
The thiocynates of which lethane is an
example are absorbed through the Skin.
They releaSe hydrogen cyanide. a power-

.


[Li] enzyme poison. in the body. Caution-

should be exercised if they are used as
garden sprays. The solutions formulated
for the control of flies are of a low order
of toxicity-for man. 
tion should bQ'ZlVOldCd.'

Since the introduction of the newer

pesticidcs- the use of the chemicals just?
'.described has rapidly decreased.

The pe-
culiar properties of the new? compounds.
together with their application'to almost
all feeds. has created a potential hazard
to health of unprecedented proportions.

Part- II

For practical purposes. the newer pesti-
divided into two main
grbups?the chlorinated. hydrocarbons and
the phosphates. din'ering'in
mode of action and other properties. both
types of compounds in sullicicnt quantity
are lethal to all animal life and most
insects.

The Chlorinated Hydrocarbon-s.-

In l939 Swiss chemists discovered the
insecticidal properties of DDT or dieliloro-
Soon afterwards

commonly

known as ienzene hexacliluride. was eon-
rtbutt-d by" France- and? the? United
Kingdom. - -

Since thiil time American ingenuity has
developed other compounds of similar
type. namely toxaphcne. chlordan.
mcthoxyehlur. lindaiie. aldrin and diel-
drin. New compounds are constantly be-
ing 

Although still undergoing investigation.

by airplanes. The unsuspecting mos-"'



- toniulogists and manufacturers had heard

 

 

responsible authorities, DDT was released
for_commercial use in 1945. Farmers. en-

of the wonderful new insectiCIdc and-
clan'i?ored for supplies. The pressure was
great. Early results justi?cdalr cxpec~
tations. .

During the latter part of World War II.
DDT had received extensive ?eld trials
in-the protection of our armed forces from
typhus and malaria. This was a calculated
rislr and there..is no doubt that the de-
struction of lice and mosquitoes radically
reduced the mortality from these danger-
ous diseases. .

In Southern Europe our troops were
dusted wuerDT powder for the elimina-
tion-of body tire. The same treatment
vigorously applied to the population of
Naples undoubtedly halted an epidemic
of typhus t'evcr.

In the tropics whole. islands were dust-

quitos and flies _werc decimated. Other
insets. bees and birds shared the same fate
to a_ lesser extent. 

An epidemic-6f gastroenteritis and so-

virus hepatitis alleeted our troops
-- iiLmanila at the-time this eity was given

problem before us.

and _with.souie reluctance on the part of

N,

4

the same treatment. The cause was never
definitely determined but was attributed
to unwashed vegetables acquired. from
'the natives. -

In the year 1950 it is estimated that one
billion paunds of agricultural chemicals
were manufactured. This gives an idea of

Physical and Chemical Properties

DDT is the least water soluble organic
compound known to science. This fact
helps to explain why chemists assumed
as events have proved-?-that
dairy cattle might be safely sprayed or
dusted with that compound. it is however.
quite soluble in many fats. Its usefulness_
as a pesticide and its danger. is to a con-
siderable extent dependent upon this
property.  - -

DDT is exceedingly stable. It is not
normally cit-rumpused by sunlight nor by
cooking temperatures. The leaching ef-
fect of rain is very. slight. Residues on eit-
i'iis fruits and friliaee in California were
found to be reduced by only 50% in 40
days. 'l'cii per cunt still remained after
90 days. In suit the compound is much
more stable. One test plot still shows



heavy contamination six years after one
application to the sail.

is a mixture of optical isomers
with a musty smell. Its application has
been found to produce an elf-flavor in
potatoes, walnuts and other crops. The
Beech-Nut Packing Company has had to
reject shipments of peanuts and vege-
tables purehascd for its line of baby foods
for this reason. With the exception of its
gamma isomer, lindane. the compound is
fairly stable. Lindane IS the most active
fraction and can now be purchased in a
relatively pure form. 

Chlordan has properties intermediate
between DDT and BHC. but on the whole
is more toxic. Aldrin and dieldrin are
chlorinated naphthalenes of similar na-
ture?wieldrin being the most toxic and
residually effective of the present in-
secticides. -

Toxaahene is a chlorinated 
of marked stability. shares
with lindane the fact that it is relatively
unstable and therefore presents less resi-
dual hazard. liowever, one investigator
has stated. that with repeated exposure,
the to:;ic1ty of lindane may increase as
much as 100 times! This 'merits confirma-
tion. -
- Toxicology

The chlorinated hydrocarbons are 
poisons and their action is associated pri-
marily with their solubility in tissue fats
or lipoitls. The cuticle or chitinnus cov-
ering of most insects is water-repellent
and seems to protect them from dessica-
tion or dcownmg. The outer layer of this
cuticle is ofa waxy nature and has a
de?nite allinity for DDT. This fortuttous
property renders most insets as vulner-
able to contact with DDT as though no
cuticle were present. And the marked sta-
bility of this chemical presents prolonged
Opportunity for such contact. lilies readily
absorb the compound through their feet.

DDT has a similar action on the ner-
vous system of insects and animals.0vcr-
stimulation of the sensory and motor
nerves iy lethal doses leads to tremors.
convulsions and. spastic paralysis. Death

from exhaustion follows. . 

Toxicity for Ian and Animals
The acute toxic oral dose for animals

this would'be about ounce. The toxi-
city_is enhanced _wlien DDT is dissolved
in dietary fat or in cils.

Suspensions and powders applied to
the skin are believed to he nun-toxic. but
in man-have occurred follow-
ing contact with only solutions.

The ordinary household aerosol bomb

- used for flies and niosquitos is thought

to be harmless. Animal toxicity only ap-
pears after exposure to a concentration
4000 times that found in this contraption.
However. Biskind has reported severe

a
I

- is about 250 nip/knreactions to these sprays, probably as a
result of hypersensitivity. ltecent experi-
ments Stillttest that repeated exposure may
lead to respiratory infection in animals.

Because DDT is poorly metabolized and

excreted 'by animals and is in: soluble, it 

is stored in the body fat. It is therefore
a cumulative poison. The amount ingested
in food may be concentrated as much as
30 times in fatty tissue. When the diet is
contamintaed. DDT is excreted in com-
paratively large quantities in bovine and
human milic. being found primarily in
the cream. Thus. cows feci thr0ugh the
winter on hay containing 8443-1 of
DDT, secreted enough of this chemical in
their milk to cause loss of weight in steers
drinking it. Their own suckling calves
accumulated DDT to-'-thc extent of 325
in their body fat. When rats were
fed for several years on a diet containing
only 1 ppm, they stored up to 30 
in their adipose tissue.

Chickens consuming contaminated ra-
tions magnify the DDT intake and excrete
it in the yolks of their eggs.

it is conceivable that a rapid loss of
weight in animals or humans storing large
amounts of DDT in their body fat couid
release enough in the blood stream to
cause of acute poasonmg. Clin-
ically this seems to occur.

At this point_it_should be noted that
fat is a living tissue which is continuously
being broken dawn and rebuilt. More-
over, all types of hodyceiis are surrounded

by semipermeable membranes contain-.

inn lipoids. which can theoretically hold
DDT. This chemical is known to inhibit
one of the oxidative enzymes. cytochrome
oxidase and may interfere with the action
of others. Therefore this type of storage

'in body fat is not like keeping butter in a

refrigerator. Toxic effects on each body
cell even with high dilution are not too
remote a possibility, 

i I Pathology

Whereas rats do not show obvious signs
or of poisoning on a ration con-
taining less than 100 of DDT. the in-
clusion of as little as 5 in the diet
over an extended period of time produces
minimal but definite liver damage. This
is reversible. if all DDT is removed from
the diet. Liver necrosis may be preceded
or accompanied by fatty degeneration or
enlargement. are resistant
to treatment with choline. While early
stages of damage may be repaired. A. W.
A. Brown states. "?liut if further chronic
doses prevent its regeneration. the detoxi-
eatiou mechanism of the liver for DDT is
impaired." .

The damage resulting: from severe poi-
with the chlorinated earl-ions varies
somewhat with the animal and the cam-
pound. However. few organs escape. Path-
ological changes reported include edema






-in oil.

  

. 

(a K3



of the brain and spinal cord. slight kid-

ney damage. necrosis of the muscles. stom-

ach and gall bladder. hemorrhages in the
heart and gastrointestinal tract and degen-
eration of the endocrine glands. TDE has

a unique tendency to cause atrophy of the

adrenal gland.

-

Acute DDT poisoning produces. in labo-
ratory animals. a period of nervousness
and hyperescitability. with excessive
blinking. cold skin. ruffled fur. lack of
appetite and muscular weakness followed
by the onset of fine tremors due to muscle
fibrillation. particularly in the heart
muscle. hind legs and back. Advanced
paisoning leads to rapid weight loss. clonic
convulsions. paralysis. and death. In ad-
dition to these. cattle and sheep develop
staring eyes. diarrhea and st. ggering gait.

of mild chronic may

include only nervousness and failure to

gain weight normally. 

Through 1951 a total of only 14 human
fatalities due to DDT were reported. Most
of these were the result of swallowing.
accidentally or with soicidal intent. rela-

tively large amounts of DDT dissolved
A few severe reactions probably-

of allergic nature?have been described.
These included two deaths. Six cases of
hemorrhagic purpura and one of agranulo-
eytosis were recently reported.

resulting: from the accidental
ingestion of DD?IF?i?clude nau-
sea. vomiting, diarrhea. nervousness. sore
throat. joint pains. muscular twitching,
impairment of sight and insomnia. ltecov-
cry-often taltes place in a few days. but

in one case took as long as twelve months.

Biskind [rem his observations has added
the following: overwhelming fatigue and
muscular weakness.
shooting pains. areas of skin hyperesthe-
sia. sense of a lump in the throat. severe
emotional depression and vague fear. and
decreased vibratory .sense.

It will be noted that many of these
are typical-of the so-called
virus infections that have been widespread
in the past five or airy-cars. I -

Bisl-iind has called attention to the fact
that the described during an
epidemic of "Virus in California sev-
eral years ago corresponded almost ?10%
with those tabulated by him in cases of
DDT toxicity. As a result of these obser-

vation?s some writers have quoted him as



peripheral neuritis.'

saying that the "Virus epidemic was
the result of DDT poisoning. Biskind is
emphatic in remarking that he ?did not.
and could not from the evidence at hand.
make such a statement." While a virus
believed to be responsible for this epi-
. dcmic has been isolated. similarity of
is conducive to much thought.

Evidence of Danger

At present the harmful effects of the
chlorinated hydrocarbon pesticides on Mr.
and Mrs. John Q. Public are debatable.
The great majority of entomologists be-
lieve that residues on crops. in milk and
?eggs and meat are low enmigh to be
harmless to humans. and that any pos-
sible toxicity is overbalanced by the ne-
cessity of these chemicals for the produco
tion of adequate food supplies. Their con-
clusions are open to question and I shall
cite 50me evidence to the contrary.

'Biskind. as one of the original physi-
cians suspecting toxic effects from the
new insecticides. has reported numbers
of cases illustrating dei'iniie reactions to
DDT. chlordan and BHC suggesting both
allergic and direct toxic reactions from
these compounds. While some afflictions

were short-lived, others perSisted for many

weeks or months.'

Pottenger noted that patients who re-
ported gastrointestinal upsets. followed
one or two weeks later by respiratory
often showed mild jaundice

and evidence of liver damage. Fat biopsies 1'

analysed for DDT varied in content from
0 to 33 ppm. He felt the exhaustion and
other associated could be at-
tributed to exposure to DDT in food or
household sprays. There was no question

in his mind but that exposure to DDT 

could cause serious illness including one
type of pneumonia-?

strongly recommends that

any foods sprayed in the process of grow- 

ing, preparation for market. or the final
handling. shouid carry labels to that effect.
Let the public know what it is buying.
Krohn_ and Pottenger in a medical pa-
per awaiting publication. and included in

- their. testimony before the Select com--
mittee of the House of Representatives

to Investigate the Use of Chemicals in
Foods and Cosmetics. conclude as follows:

(I) The widesprcad'_ use of chlorinated
hydrocarbon insecticides has produced
many cases of toxicity. -

(2) These insecticides accumulate in fat
and principally damage fat containing or-
gans. as the liver and nervous tissue.

(3) lit the presence of characteristic
signs. and blood studies. dqo
tection of chlorinated hydrocarbon insecti-
cides in a biopsy of the patient's fat _con-
firms. the diagnose: of insecticide 
ing.

(4) Chronic toxicity from these insecti-
- 

is

23331??

up};

.r I

cides has been observed more frequently

than acute toxicity.

The writer has observed a small number
of cases with neeative fat biopsies but
evidence of liver damage. that gave a sig-
nificant history of exposure. He has also
seen several cases of indisputable allergic
reactions to these substances.

In other words. the presence of DDT
in body fat proves that absorption of this
chemical has taken place but does not
necessarily indicate actual poisoning from
that substance. 

However the presence of liver damage
favors such a supposiLinn?particularly if
more than 5 of DDT :5 present in
adipose tissue.

Contrariwise. an allergic or'hypersen-
sitive individual may theoretically ex-
hibit violent reactions to minute amounts
of DDT without absorbing measurable
amounts. The only criterion necessary for
diagnosis is tlte reproduction_of symp-
toms upon repeated contact.

Sources ot Exposure
There is adequate evidence to prove that
many foods raised commercially contain
at least traces of DDT or Similar com-
pounds._ These include staple vegetables
and fruits. 
For all practical purposes DDT does not

penetrate thr0ugh' the skin of fruits. How-A
everE it becomes incorporated in the shin 
or mud and can not be removed there-"

from by any washing method known. In
spite of the loss of nutrients present in
this layer the only safe method of avoid-
ing possible high contamination is to
pool all fruit. - .

Beef. chicken. lamb and pork may con-
tain relatively large amounts of DDT if
the fodder of these animals was containi-
natcd with the substance. The use of corn
huslts. pea vines and other heavily tainted
sources of livestock feed renders this pass

sibilily a real one. Fatty portions of meat.

contain the largest quantities. 

As has been mentioned before. milk."-

ereatn. butter and eggs may be serious
offenders if contact with sprays or DDT
containing food is adequate. Nor is mar-
garine innocent. The vegetable fats in this
product may be contaminated as a result
of spraying or dusting.

It took many months before it was rea-
lized that dairy cattle sprayed for fly con-
with DDT or in low dilution
would these substances in their
milk for ten days or inure. 'l?his even
occurred when barns were foimcd or
whitewashed with DDT spite of the

fact that food bins we're covered and the'

cattle removed. At the present time these
chemicals are not recommended for use
in dairies or. on beef cattle. instead. lin-
daiie and are. suggested as

substitutes._

.4.

ironically enough. flies in most areas
are becoming resistant to DDT. And what
is more?once resistant to this chemical
they rapidly seem to acquire immunity to
others. Authorities therefore. are once
again stressing the old methods of control
?cleanliness and eradication of breed-
ing places. And so we come full Circle
once more. -

While translocation of DDT {rem the?
ground or foliage to the edible portion of
plants seldom occurs. enough BHC and
lindane may be so transported to give an
off-flavor to potatoes and other vegetables.
Therefore the use of these chemicals is
decreasing.  .

Exposure to DDT in the home is all too
common. Aerosol bombs for control of
flies and fleas should be considered un-
sa fe?as should citlordan powder or sprays

for the destruction of ants and other pests. 

A case of poisoning from the latter com-

'pth was recently-reported in the

J.A.M.A.

The increasing use of lindane. DDT and .

similar compounds in restaurants and'
other public places by vaporization has
been mentioned in a previous report and

is.attended by definite hazards. -
individuals may incur 

damaee'from contact with outdoor sprays

used domestically or commercxally. They

should be .avoided. .
Tentative Conclusions'

The purpose of this article is not to?

create-alarm bill to alert thoughtful per-
sons to the hazards inherent in the use
of DDT and allied substances.

At the present time food production

-in the United States would be disrupted

in a few weeks it the newer pesticides
were discarded. Tins would be disastrous.

However. 'he evidence herein presented 

points out inherent dangers which were
not forseen. and if suspeeied were and
still are - ignored by many of those to
responsible positions. More evidence of
human toxicity is needed.: . a. I
YourJlousc Select Committee is doing
a fine piece of investigation and deserves
encouragement.

PART 

'The Organic Phosphates
These compounds were developed in
Germany durini: the past war for use as
insecticides. Rumor has it that they may
he the so-called ?nerve eases" whose po-
tentiality for harm brought bad dreams to
our intelligence staff at that time. lie that
as it may. these products are so tusie that
they are limited to applications in the
field by trained operators. Their use on
livestock and pets is not recommended.

(Department of Understateinent.)

Physical and Chemical Properties

The orizanie phosphorus compounds are
among the most toxic chemicals used for

raaztr1



pest control. Exposure to relatively small
amounts may result in severe illness or
death.

TEPP (or is
the most potent ol' the group from the
acute standpoint. it is several times more
toxic than nicotine. 
tetraphosphate) is on a par with nicotine
and owes most of its destructive potential
to the perCentage of present in the
mixture. Both these compounds are ra-
pidly hydrolysed and therefore dangerous
lor only a few days after application.
They are also rapidly detoxified in the
body and therefore not stored in the

tissues.

Parathion is an ester of thiophosphoric
acid and contains a sulfur radical. It is
more slowly decomposed and as a result
presents more of a hazard. It is also fat

soluble to some extent.

Toxicology

There is little difference in suscepti-
bility among various animal species to
the toxic effects of these compounds.
Several hundred human cases of poisoning
have been reported with a number of
fatalities?most of them due to careless-
ness in handling the products.

The organic phosphates are toxic by
absorption through the skin, by inhalation

and by ingestion. As little as- one drop of .
concentrated solutmn in the has

caused death. Whereas and 
have failed to show cumulative effects.
animal experiments have shown that re-
peated contact with small amounts of
parathion may produce poisoning.

It has been estimated that 25 mg. of
TEPP by mouth will produce moderately

severe in man. 100 mg. orally-

would probably be fatal as would 500 mg.
applied to the skin. .
The all-tyl phosphates inhibit the enzyme
cholinestcrase. This enzyme destroys ace-
one of the chemicals liberated
at nerve junctions and endings to transmit
the nerve impulse. Thus a lack of the
enzyme results in a marked prolongation

of each nerve impulse affecting primarily'

the parasympathetic part of the nervous
system. \licotine-lilte efteets are also pro-
duced.  7 7

On acute exposure develop
rapidly once the cholinesterase level has
been sufficiently lowered. They include
loss of appetite. nausea, vomiting. abdom-

inal cramps, exec-ssch sweating. papillary 
constriction. diarrhea. respiratory diffi-

culty. headache. dizziness. muscle twileh-_
ing and weakness. Severe cases develo
coma. convulsions and death. 
Repeated exposure to small dosvs may
result in illness of severe degree. The en-
zyme system recovers fairly rapidly after
one episode. but may take weeks after

 numerous subclinical contacts. Testing a



tion enters the picture. -
Susceptibility to the organic phosphates 
is variable as one would expect. in cases 

blood sample for cholinesterase levels may I

help in diagnosis. Sedatives and the intra-
venous administration of atropine in doses
of 1 to 2 mg. as often as every hour may
be life saving in severe cases of poisoning.
Death may occur within six to ten hours
of the development of Recovery
from acute attacks is usually complete.
Adequate protective clothing and special
masks must he used by those handling
these chemicals.

TEPP and 1181?? present no problem.
Fruit pickers have become ill after enter-
ing an orchard treated Willi parathion
several days after its application. Food
crops are not suppOSed to be sprayed

within thirty days of harvest. A safe resi-

due on any one item of diet is censidcred
to be 2 of parathion. In the case of

'citrus fruit the peel will absorb and re-

tain higher residues than are allowable.
This is also true of olhcr_fruils. :?ts re-
gards apples and pears. if parathion is

applied?"stricin in accordance_ with the 

recommendations of the Bureau of Ento-
mology Plant Quarantine of the U. 5-.
Dept. of Agriculture with particular refer-
ence to the time between the last spray-
ing and the harvesting of the fruit. normal
weathering should result in parathion
residue no greater than a fraction of a
part per million." 7

Nevertheless toxic parathion residues
have been reported by Pottenger in bread
and canned fruit. Again the human equa-

of suspected poisoning a physician should
always be called. Blood tests may uncover
chronic cases. -

Absorption and Translocation
TEPP is rapidly?absorbcd into plant
tissues as shown by growth inhibition or
stimulation and other metabolic disturb-
ances. The rapid breakdown of these com-
poands limits the importance of their
absorption to the effect on plants.

Parathion-has been reported to kill a

. variety of insects on plants when applied
only to the soil. Biochemical studies and 

bioassay have shown that absorption and
transloeation does take place in insecti-
cidal amounts. The relative stability of
this chemical together with its toxicity
makes these findings of great importance
to animals and humans. Chemically al-
tered parathion has beeniound in some

citrus fruits.. The toxicity has not yet

been reported.? I
systemic Insecticides
These are

lranslocated by actively growing plants
*in sufficient amounts to kill insects feed-
ing at a site distant from the original ap-
plication. The principal compounds of this
group are 

 

chemicals absorbed and-

L-

-. 

.


- 







phoramidel. "Systox" filictliyl etliylmer-
triphosphoric
acid penta dunetliylamide and the his
fluorophospliine oxides. .

After absorption the chemicals may he
changed to less or more toxic compounds.
Injury to the plant is possible. Transloea-
tion to edible portions may make them
toxic or produce a tainted Hat-or. Al the
present time theme of these svsiemic
pi-stiCides on food crops is not permitted.
They are still in the experimental stage
and should always remain so as far as

food for man or animal is coucerned.

It should be stated once more that in
addition to the systemics just described.
evidence of absorption and translocation
exists for arsenical compounds. parathion,
benzene hexucliloride. DDT. tosapnene.
chlordane. dicidrin. selenium comnountls.
fluoroacetates. and fiuorohyn?rins. The last
four groupshavc been considered too toxic
for use on food crops. dicldrin
has recently been released for use on
peaches in California. Benzene hexa-
chloride (as reported in Modern Nutrition
for October) may no lancer be applied in
vegetables or fruits in this. state. lime.
over, where it has been used in the past
the soil may remain cantaminatcd to a
decreasing extent-for five years or more.

It has taken a long: time for the au-
thorities to take Such a step and now
treated ground will not be free from
traces of the chemical for eight or ten
years more. In the meantime the popula-
tion has been exposed to the effects of
BHC for a period of four 0r five years.
This is but one example of the price the
public is paying for the use of new chem-
icals in and on our food without adequate
investigation of possible harmful effects
from these paisons.

Practical Aspects

By this time those who have followed
these articles should be considerably
alarmed over the number of pmsons hu-
man beings in this atomic age must en-
counter. And so! It is a deadly and
grim situation. Naturally the question
now arises?so what? Where do we go
from here?
family. myself and our \vciniierftil Nation
from what seems to be serious harm?

Unfortunately there is no pat answer.
However. I am going to discuss steps
which seem imzical at the present time.

Legislative Action 

While your government representatives
in local. state and rt?llPl'Ill real-
lae llIt?l'L? is a problem and wish to pro-
tect the public, they are under. pressure
from pesticide mantit?acturers and dis-
tributors whose chief interest naturally is
in sales. These tzrt-ups refuse to believe
that exposure to sinail amounts of insecti-
cides constitutes a great threat to public

health.

What can I do to protect my-

Entouwlogists being primarily rcspon-'

sible for the production of large crops to
feed a hungry world are loth to accept
proof of chronic toxicity?anti such proof
in humans is difficult to obtain. t'l?here is
plenty of evidence of such potential haz-
ard from animal experiments.) Meanwhile
Departments of :?iei'iculttire are busy con-
ducting investigatmns along lines of ?010
icity for crops, animals and man.

You are the guinea pigs and must write
your state and federal representatives
asking them to take action. You must 3150

contact your local departments of health 

along the. same lines:7 The other side of
the question m?ust be heard from.

Personal Precautions

in an attempt to avoid contaminated
fonds one must not go off the deep end
and develop malnutrition in the process.
Many foods from large truck farms where
spraying directions are carefully followed
and supervised contain no detectable resi-
dues. However. a certain number of crops.
because of ignorance, wdlful neglect of
:?rlvice. drift from nearby fields or other
show slight or moderate and oc-
casmnally, heavy contamination. Surveys

in the Les Aneelcs area in l950 showed

detectable residues of DDT in more than
half the samples examined. About four
percent were heavy. Arsenic contamina-
tion was negligible This risk is unpre-
dictable and at the present time cannot
be prevented. The following suggestions
may be helpful:

I. Whenever possible. purehase' foods
that have been grown without the-use
of insic insecticides. At present the supply
of such foods is markedly limited. Encour-
age the development of organic farms in
your area. Grow your own vegetables.

2. Do not us: sprays or aerosols in your
house that contain DDT. Chlordane. iin-
dime. BliC Ior Maire sure
that commercial companies

do not use these chemicals in your house 

or on your trees and shrubs. Do not eat
in restaurants usmg vaporizer: for fly
control.

3. Peel all fruits and vegetables that
lend themselves to such treatment. Scrape
the outside layer from celery. Washing
will not remove residues. l?urchase ones
from local ranches that are not using DDT.
UllC or chlordane in hen houses. (Metho-
xyt-hlur and lindaiie are not cumulative
and do not ordinarily iitiild up in chicken
tissues. hence in cans.) -

Milk. cream and butter are much safer
formerly. Must (lairii's now use

methuxyehlnr and lindanc for fly control

and very little is excreted in the milk.
Cni-i-k with your local dairy to find out
what chemicals they use. Shift to another
dairy if yours is ?ll? tislni: DDT. BliC or
chlordane or tlieldrin by the "strip"
method. 

988'le 

 .Vitamin supplements such

4'


- - - .


Most vegetables growing in the ground

such as potatoes. carrots and heels are 

comparatively safe. Contamination by
Ill-1C will produce an oil flavor and thus
give warning. Potato skins may be eaten.

Fruits and vegetables that have at in-
tervals shown contamination with DDT
in the Los Angeles area are celery. cab-
bage. cauliflower. greens (spinach. par-
sley. endive. turnip. etc.) and lettuce.

.Apples and torriames are included, but
quite safe when peeled. _ln case of stom-
ach Upsets it would be wise to avoid these
vegetables for a few weeks.

Dried fruits may be higher in DDT than
fresh fruit and they cannot be peeled.
Some olives and grapes are suspect. Prunes
show very little residue as a result of the
alkali treatment given them.

Moderate amounts of salads greens are
fairly safe and desirable food stuffs. Home
sprouted soy beans, mum: heans. alfalfa and
the like from untreated seeds provide an
excellent source of uncontaminated and
nutrilious salads.

Lean meats are satisfactory sources of
protein and sea foods are naturally free
of any resndues. . -

4. Attention to other factors' affecting
the general health is important. Adequate
rest. avoidance of worry and tension. mod-
eration in the use of coffee. alcohol and
tobacco, sufficient exercise and a diet low
in refined carbohydrates need emphasis.
as yeast or
vegex. dess'icated liver. rice bran con-
centrates. fresh wheat germ. cod liver oil
or its concentrates and kelp for trace min-
erals are helpful. More potent prepara-
tions are often indicated but should pre-
ferably be taken only under medical super-
vision. - -

Allergie.states and chronic infection of 

any sort interfere with adequate metabol-
ism and should be corrected if possible-
Naturally other ailments should receive
the necessary care. 

5. Repeated attacks consisting of up-
per respiratory infections and/or gastro-
intestinal together with loss of

weight and marked fatigue- suggest pesti:

cide toxicity or allergy. In any case of
this sort see your physician. Other causes
may be Operative. If he finds nothing
wrong suggest the possibility of insecticide

poisoning-and ask for a bland count and


4

- liver function tests.

Do not try to diag-
nose anti treat yourself. You are almost
bound to be mistaken and to do more
harm than good. -

In? de?nite cases of DDT poisoning a
strict diet together with other nutritional
therapy will be necessary. Recovery is
often discouragimzly slow but. to he expected
once the cause has been recognized.

Summary and Conclusions

The various types of chemicals used for
the control of insect pests have been de-
scribed together with their potential tox-
icity for man and animals. The fact that
some of the chlorinated hydrocarbons. even
when absorbed in trace amounts. may be
concentrated at least thirty in the
body fat of animals and human beings.
renders even slight contamination of our
feeds a public health problem. of first
importance. Such contamination is pres-
ent and in susceptible individuals is pro-
ducing illness. This fact must be faced
and steps taken to protect the public in-
terest. The Delaney Congressional Com-
mittee has been investigating the problem
and will recommend legislation if this
seems needed. The Committee needs the
support of all. 

Suggestions have been made by means
of which the public may reduce the intake
of insecticides to a minimum. It is impos-
sible to avoid all contact.

If the use of insecticides ivere'suddenly
stopped crop failure and hunger Would be
widespread. Perhaps the solution may lie
in the following steps:

ll) Fairly rapid abandonment of the
more toxic pesticides in favor of less harm-
fut ones. 

(2) The development of widespread pest

control by naturalencmies. Excellent re-'
search on this is being carried on-

at the Riverside Experimental Station.

(3) As Pottengerdies-suggested, foods

that have been sprayed or dusted should
be so lahelled including the insecticides

what it is buying. .

used. The public has the right to know.

Testing of food crops before market--

ing. While costly it might be cheaper in
the end. to test for BHC. DDT. arsenic.
lead and parathion. where any or all of
these have been used on food crops be-

fore the crop is sent to market. To allow 

for differences in concentration and other
factors. a sample front each acre might be
indicated. Thus heavxly contaminated

crops would be discovered before reaching -

the consumer. .

- (5) The passage of State or National
laws condemning the manufacture of
aerosols. sprays or vaporizers containing

DDT. chlordane .lindane or 

for use in homes or public places. 'l?hes:
chemicals should be used only.outdoors
and then with extreme caution.

(6) The gradual spread of organic farm-
ing: including the use of city
tvastes for fertilizer purposes. The work of

Pfeiffer along the lines of special cul-

tures to produce a rapid breakdown of
organic material shows much premise. In
spite of a great deal of skepticism. an in-
creasing number of fat-titers and soil ex.
perts believe that healthy plants are more

-. -.- ..

-M (1/1

933311;

An? -- -bu-o-

-.

-4. 

resistant to insect pests and that healthy
plants can only grow in rich soil. It is
probable that the future of America and.
indeed of the world. depends upon the
rebuilding of our impoverished soil
through the addition of organic matter.
trace minerals and judiciously used com-
mercial fertilizer?not upon the develop-
ment of newer and more powerful insec-
tieides.

2901 ??ilshire Blvd.. Suite 345



References

BISKIND. M. 5.: Statement before the Select
Committee to lnveslnzate The Use of Chemicals
in Food Products (Dec. 12) ltifio.

BISKIND. M. S. and Irvine Bieher:
tll. No. 2 (Aprili Isl-tn.

Louis: Statement before the Select
Committee to Investinate the Use of Chemicals.
tn Food Products (May) tail. 

BROWN. A W. A.: Insect Control by Chemicals.
39'9" New York: John Wiley .5: Sons. .lne..

a .

CASIDA. E. and T. C. ALLEN:
and Translucalion of Insecticides by plants.
Agricuit. Clicin.. Vol. No. ti tJunei I552.

COUNCIL on PHARMACY AND CHEMISTRY:
J.A.M.A.. 1491.1?? li-lay 

Am. Jl.

Absorption

on PHARMACY AND CHEMISTRY:

Pharmarotocy and toxic-omit)! of certain phi-s-
pnortis insecticides. tf-i-pt.
9t 1530.

John: Statement before the Select Com- -

mittee to lnvesueate the Use of Chemicals in
Food Products Iliayl lt?l?il.

EDITORIAL. DEPT. Insecticide storage
in adipose tissue. 145: 135-136 til-far. l0l tail:

W. M.: Deposit and residue of recent
insecticmcs resulting from various control prac-
tices in California. Journ. Economic Entomol-
-

KNIGHT. G. R: Testimony before the House Select
Committee to the Use of Chemicals in
Fonda and Cosmetics. Part 2. 193i.

KROHN. Bernard: Statement before the House
Select Committee to Investii'ate the Use of
Chemicals in Food Products. Part 2. 1951.

LANG. E. P.. F. M. NUNZE: and C. S. PRICKETT:
Occurrence of DDT in human int and milk.
A.M.A. Archives Ind. Its-g. Occ. Med.. 3:245-
216 tMar.) lDSl.

A. J.: Pharmacological consideration
ff insecticides. J.A.M.A.. H-l:l0-l-tOB IScth

LEIIMAN. A. 1.: Bulletin of the Association of
Food and Drug Oti'icials.of the United States.
Vol. XIV. No. 3 (Julyt 1950.

LEIIMAN. A. J.: The Major toxic actions of in--
secticides. Bulletin N. Y. Acad. Med.. 25:332-
331? tJunc) l?-lD.

A. J.: Some toxicological reasons why
sortain chemicals may or may not be permitted
..s tot-d additives. Bull. Assn. Food and Drug
Officials of U. 5. Vol. H. No. 3 _lJul)'l IDJD.

LEMMON. Allen 3.. Chief. '?ureau of Chem..-
Annual report for the calendar year l950. The
Bulletin. Dept. Aerieult.. State of. California.
Vol. .14. No. t. 1950.

It. Mode of Action at Oreanie

Insecticides. hat. Research Council. Wasti..
D. C.. lit-la.
PM. Jr.. Testimony before the

Home Select
Chemicals in Foods and Cosmetics. Part 2. 135i.

l'tOllWEl'l. 5. A. and H. L. ila'llct': Pharmacology
and toxicny of certain pnusimc-i'us insecticides.
mam-ma (Sept. 9t 1950.

Select Committee to tm?eshcatc the Use of Clicin-_
mats in Food Products: Report No. 3254. 
Congress. 2nd Session. llouse of Representa-
tives. Union Calendar No. 1135 ?an. in tail.

SIMMONS. S. W.: llcalth hazards of Economic
poisons and related substances. Am. J. Trop.
Med.. Vol. No. 4  (July: 195]. I

- . ADDENDUM 7: i

In reprinting' thisi'article. which first ap-

Eearcd 20 years ago. it seems logical to
rielly review what has happened since that.
time.

Chemical contamination of our environ-
ment has Iii-come obvious. DDT and other
chlorinated hydrocarbon pesticides have un-
expectedly been spread by wind currents
thouglimit the world: They have penetrated
our food chains and are now found in high
concentrations in some ?sh and seals. bulb in
the Arctic and Antarctic. Many of our rivers
are sewers and some of our lakes are dying
as a result of changes caused by chemical
ef?uents. Our Coastal waters are so badly
contaminated that in some art-as many liin
have lu-i-n [mind with cancerous growths.
As a result ul' "It? brown pi-lii'ttn niny
simii be extinct and other birds are seriously
lhrt-alvni-il.

Human beings carry in their fatty tissues
about 12 partsti-r million of DDT or its
degradation products Many individuals
have measurable residues at other eiiluri-
natcd hydrocarbons. includint: the pin-nutty
herbicides and the polyehlurinated bi-

It



ph?enyls (PCBs). Degenerative diseases. in-

cluding cancer. are on the increase. --

Studies by the Bionetics Laboratories in
196:": but not released until lQii? 
showed that the pliennxy herbicides. 
Bali?'1?. 2.1l-Dl? 'and Silvex not only pro-

duced birth defects but cancer in experi- I,

mental inice.'. So did DDT and related
chlorinated hydrocarbons. as well as other
herbicides such as the Trinziniis. l?artiquat
and Zi-t'trzin. It is of considerable concern
that DDT and have been user! on our
food crops since about 194:3 and that 2.4-0.
Silt-ear and other herbicides have
been available for use on lawns also during
this period, Since :inrl Silvr-x are
contaminated by the 2.3.7.3 
Iii-tizn-par:i-diusinn. in tiniimtl experi-
im'nls. Irma: In be up to mu: 

firm-s iitori.? orfir'i: Him: Hirrlt'rlmnirfi' in prii-.

rim-in: young. ll)?
flu-so Itcrtu'i'iifrs "my be much 
flimi Misfit-civil. Incidentally. the toxic climb;
of the were 
triisively documented by M. Jacqueline. Ver-
relt. i?li.D.. Division of 'l'uxicology. F.D.A..

Committee to the Use of.

Ainw



irnApril of 1910 Obviously, the results of
animal experiments cannot be necessarily
applied to human beings. Nevertheless. the
results are of great: importance since minute
amounts of carcinogenic chemicals may. at a
much later date. produce canCer in suscepti-
ble individuals or animals. According to the
Delaney Aint-tutnu'nt to the Food. Drug and
Cosmetic Act of HMS. any chemicals which
protltn'e cancer in animals must be banned
from foods or cosmetics.

'l'henoxy lierhicides .evcn though in trace
amouan. now contaminate about thirty per-
cent of our water supplies and a recent
report indicates that the ubiquitous l?CBs
have been detected in fifteen to twenty per~
cent of our drinking water. Such contamina-
tion may occur from local applications.
ef?uents of various types. drift from aerial
Spraying. transport by winds from other
parts of the country. or by as yet unknown
means. 

"Agent Orange." .1 mixture of equal parts
of and was used for many
years as a dcfoliant in South Vietnam. This
chemical weapon may have irreversibly de-
stroyrd the balance of life in the estuaries.
as well as ruining as much as thirty percent
of' their hardwood forests. Not many indi-
viduals are aware of.the fact. that "Agent.
Orange." contaminated by the fantastically
potent tctrachlorodihenzo-p-clioxins, was
used for many years in our own forests be-
fore it was sprayed as a defoliant in Viet-
nam. Since it ms banned as too dangerous
in Vietnam in 1970, why hasrits use not. been
forbidden anywhere in the United States?

r. Director of the Environ-
mental l?rotection Agency. is attemptingr to
limit. the use of dangerous chemicals. such

as DDT. Silt-ex anti similar chlori-l

nated hydrocarbons. in order to protect our
environment. He needs support. Powerful in-
terests are opposing this move. Proof of
de?nite damage is now at. hand through the
repeated pesticide spraying of Globe.
Arizona. and the resulting extensive research
of one of the victims Billee. Shoecraft 
whose book "Sue the Bastards!" provides a
cogent and moving account of her experi-
ences. in the Globe area. plants and crops
were destroyed: domestic and wild animals
died or disappeared :human beings and ani-
mals became ill; birth deformities were noted
in animals. as ?'I'li as human beings. Repro-
ductive dif?culties atTi-ctcd women amt some
of them long past the menopause much
to their consternation: bled from the

uterus. Recurrent attacks of "virus" were

frequent. - .

In 1971. the flureau of Reclamation used
Silver: on the ltiu titan-hr lliver near San
Acacia'. about sixty miles south of Alim-
qtlerque, in an attempt to clear salt-hush
front the river banks The Burr-nu felt. that.
this was usint: too water. prior
warning. helicopters sprayed the area. run-
turninatirn;I in the process members of two
families and two herds of cattle_ Both the
mount beings and the cattle became ill and

many of the latter died. The remainder of
the herds. even though they hart been re-
moved from the area and were eating vora-
ciously. lost weight and had to he sold at a
tremendous loss. Post-mortem examinations-
indicati-d damage to the liver and intestines.
as well- as to other internal organs. Thus;
we know that Silver can not only cause se-
rious illness in cattle. hut death as well, if
the exposure is great enough. Fortunately.
the human beith recovered.

is contaminated by some of the di-
oxins and,other noxious compoands. A re-
cent. study. reported to the French Academy
of Sciences. involving the use of pheasant
eggs injected wirh revealed startling
findings: I-latchability was reduced by about
70 percent and numerous embryos were
deformed.

Mast alarming was the ?nding that in
fifty pert?ent of the-emhryos. "the genital
tract of the male embryo reSemblerl that of
a normal female." The authors is
important to note that. even the vapors of
2. 4-D have an analogous action . . . \Vhen
non-treated eggs were put in treated nests.
the embryos show the same sexual anom-
alies." Conclusion: ?It is a question of fem-
inization or physiological castration." (Are
human beings exempt from this in?uence?)

Disturbing reports of cancer developing in
Swedish and 2.4.5-T
alongr raiInwarfrights-of-way and of deformed
babies being born to two women in Australia
exposed to have apparently had no
inlluence on those who are using these com-
pounds in our forests and on our ranges.

Californians should know that for the past
twenty years, 2.4.5? and]
or_Silvex have been. and still are being used
in variOus combinations in. our national for-
ests up and down the coast. in order to con-
trol weed growth along firebrealts and power
lines. as well as to destroy brush in planned

brush conversion areas; \Vhether spraying_

is done by helicopter or ground crews. drift
is inevitable and this may account for
the fact. that many Californians are shewing
the presence of these herbicides in their
blood.- Whereas herbicides are being used

atong.sL-zte highways. the application of.

anrl Silvex have not. been authorized
by the State of California for three years.
Compounds being used include Simazine
and smaller amounts of Atrazine and l'ara-
quat. It is. therefore. probable that most
of the phenoxy herbicide contamination is
the" result. of air-borne residues; This needs
more study. 

The extremely high. incidence of virus
in experimental animals eat-
posed to pesticides during the 
Studies for terntogenit'ily and carcmogentc-
ity is highly significant. - .

in tin: past few months. an 
number of illnesses 511??! to be associated
with ln-rhicitle spravimz in the Les Anm?lt?a
at 'l'ln-se Su-Citllt'il "virus infections" tend
to recur in waves and are 
toting. Hall a dozen patients have said they

. I

.



felt they were dying. These occurrences are
extremely disturbing. .

Same specimens of heel contain residues
of the estrogen. 
which has cancer-inducing properties. even
in minute amounts. Meat. and particularly
liver from these creatures. mav also contain
residues of the phennxy herhit'ides and other
chlorinated pesticides. Chickens supplied
with food containing stillnestrol. of course.
may also exhihit residues in their meat and
livers, Just. recently. the use of stilbestrol
in animal feeris has been' harmed. llowevcr.
pellets may still he implanted subcutaneous-
ly in animals. and \'-'hcrcas the residues will
prolrahly be less. there is no proof that
some will not remain.

Lindanc vaporizcrs are heing phased out
but are still helm: advertised and sold. "No
Pest" strips may he just as dantarrous as tho
vaporizers. f'l'wo monkeys who chewed on
their flea collars containinr: dichlorvns. the
active ingredient of many "strips." were re.
ported as suffering from repeated attacks of
virus pneumonia together with other com-
plications. (toe of them died and the other
reenvered after a prolonged illness.)

sponge baths for infants

were recently found to cause serious poison-
ing and a numlter of deaths. A three percent
Solution of hcsaehlorephene had been used
and ti. is now Lnown that: the scrotal area
absorle practically 100 percent of most
compounds applied to it. 
has also been shown to contain other toxic
contaminants. sut'it as the ridden-antiinxius.
has now her-n banned for
use In tlrutts and cosmetics. est-cpl; on pre-
scription. This is another example of long
delay In. recognizing toxic effects from the
chlorinated 

Last year. liquid formulations of 
were banned. together with any applications
near water supplies. campgrounds, etc. On
June 14. 1972. with a few exceptions. the
use. of DDT in the United States was ordered
discontinued by the Environmental l?roiec-
tioo Agency as of December W72. This
is as it should he. If it 

1 Suspect that so-t?allerl "virus
infections? which have appeared with in-
creasing frequency over the years and which
produce such as nausea. vomit-
ing. diarrhea. prostration. severe headache.
hotly pains. depression. sweats. low grade
fever, sinus congestion. sore throat. laryn-
gitis. hoarseness. feeling of Constrictinn un-
der the "virus pneumonia." and

.severe pains in various parts of tin: body

may all he due to contact. with pesticides
in our air. fund and water. eittn-r acting as
Virus per se. or lowering resistance In iruses.
(Morton llisl.ind. MJL. \?l?estpm't. (Touucc-
ticut. speculated on this fart years ago.)

The suggestions in my original article are
still Valid. However. since the Mtl'tjl?t'l of
memory has surfaced. sword?sh were found
to he hinher in tnr-reury content tl:.m other
fish. They were. therefore. haunt-d tempu-
rarity and ?renew carefully screened so

a

that swordfish appearing on the shelves is
fairly safe if one does not eat It more than
once weekly. Insemination or museum spece

mom; of various types of ?sh has shown the -

presence of mercury in those which were
twenty. thirty or forty years old. This indi-
catcs that. the importance of mercury has
not yet been completely evaluated.

Lead contaminates the air of our cities. as
do fluorides. nitrous asides. etc. A good in-
take of Vitamin helps to reduce lead
toxicity. Vitamins A and  together with C.
help to minimize srnn: damage. to the lungs.
Liver. yeast and B-compiex in addition to
these vitamins and a good diet. high in raw.
uncontaminated foods grown. infertile soil,
constitute logical nutritional insurance
against infections and the development-of
decenerative diseases including cancer.

Obviously. resistance to toxic chemicals is
related to active enzyme chemistry which.
in turn. depends upon good nutrition and
avoidance. insofar as that. is possible. of all
chenncal compounds which are enzyme

poisons.

Fluorides are now one of the most serious

'cmuaminants of our ecosystem. They are

producing allergic and toxic reactions.
'l'hereinre. the endless drive by the U.S.
l?nhiic Health Service and the American
Dental Association backed hy our tax
dollars to promote arti?cial ?uoridation
of water supplies. seems not only stupid and
fooiliardy hut dictatorial.

Unfortunately. other toxic herbicides.
aside from the pitenosy are stiil
being; used for weed control on vacant lots'
and alon: highways and freewzys in Les
Anecles County. as well as in agricultural
areas. These include Diquat. l?arar;1tat. Atra.
zine. and Sirnazine. (l?arar;ttat possesses the
unique quality of causing lung damage when
it is inhaled; swallowed. or applied to the
skin in sufficient. quantity.)

Research funds are urgently needed to 

monitor foods. water. plants. and people for
the presence of organic phosphates. herbi-
cides anrl other chlorinated pesticides as well
as lead. arsenic and mercury. Twenty thou-
sand dollars is needed for an adequate tn-
vestization. Any donation will help to keep
the work moving until larger gifts are re-
ceived. Checks made payable to the Price-

l?otlenger Nutrition Foundation. lahcicd for
_"rescurclt purposes and to me.

will be most welcome. Such donations are

deductible for income tax purposes and will

he acknowledged. need your help!
Once again. I would :nlsazest. that. you look
for sources of funds: grow
vegzt-tahh-s in your own backyard: matte your
opinions felt hy writng your legislators; if
our civilization is to survive. the. use of
putt-rd. and persistent. lh?hltCitlt'S must he re-
tillt't'tl tu an alu-ulute minimum: mineral and
or;::mic Inattcr returned to our soils and
lnutl grown for "rptality" rather than
?quantity.? as reennunended many years
aizn try A. Albrecht. Chairman.
Dept, of Soils at the University of hlissouri.





 In?.lvYour future and that of ymar family do- be controlled if our food supply and health I f.

pcnd_upon yom :Iciinns. are to he .-..
. acu'nty Is Inmorinnt. gram} autumn-Is more - "Vcrhum saptcnu sans est."  

effective. SopMurn Califqrmang ?luglgId ch- . - .
tact and mm :m hulnpon bo- 

cicty chaptrr. 'l?iu-ru arc chapters Pasa- Unl?t Int Ilium-ml: Cn?ginnsl?iog? .
. . . .. run. . ow . .1 . nc.. 1 . t. -

gcna? san drs?sr?tuxhh vaucii ll. Conant-er natureSimon ShuNll?f'. 1571. 53.95 -
snununns may rc?qmrc M. L-  Clucmca} ratings.

the us_c 9f pang-Isles and lhc_ju:llcious 
 of a?tl?iclnl the 1n. Fag?; ?35  3mm?; 
discriminate use of these subsmecs mus: --

. 

8211 Moo




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