research awi! jory board - international joint … · following report on its findings prepared by...

RESEARCH AWI! jORY BOARD

HEALTH IMPlICIITIONS OF H1A

I

REPORTTO THE

OF THE INTERNATIONIIL JOINT COMMISSION ON THE HEALTH IMPLICATIONS OF N'TA

TASK FORCE

MR. I! 0. FOIEY, CHAIRMAN DR.E.BLKlN6 DR. J.MUUER DR. R.II.GllYER OR. H.1.RlK OR. N. CIERNOA

MAY 1177

l NTERNATIONAL JOINT COMMISSION

GREAT LAKES RESEARCH ADVISORY BOARD

May 1977

Great Lakes Research Advisory Board

I n t e r n a t i o n a l J o i n t Commission

Canada and t h e United S t a t e s

Members of t h e Board:

The Task Force on t h e Heal th Impl ica t ions of t h e N i t r i l o t r i a c e t i c Acid (NTA), a s a requirement of i t s Terms of Reference, is submit t fng t h e fo l lowing r epor t on i ts f ind ings prepared by t h e members.

Respec t fu l ly submit ted,

Chairman %I

G. Becking

'i L ,

N. Chernoff

$!L~-' H. L. Falk

R. A . Goyer

Page

PREFACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . CONCLUSIONS 7

. . . . . . . . . . . . . . . . . . . . . . . . . . . . INTRODUCTION 9

TOXICOLOGY

. . . . . . . . . . . . . . . . . . . . . . . . . . Pharmacology 11 Reproduction and F e t o t o x i c i t y . . . . . . . . . . . . . . . . . 11 . . . . . . . . . . . . . . . . . . . . . . . . . . Mutagenici ty 13 . . . . . . . . . . . . . . Acute. Subacute and Chronic Toxic i ty 14

CARCINOGENICITY

D e f i n i t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 . . . . . . . . . . . . . . . . . . . . . . Experimental Design 14 . . . . . . . . . . . . . . . . . . . . . . . . . . . . Resu l t s 1 7

. . . . . . . . . . . . . . . . . . . . . . . LEVEL OF HUMAN EXPOSURE 21

RISK ASSESSMENT

Dose Comparisons with S p e c i f i c Carcinogens . . . . . . . . . . . 23

BIBLIOGRAPHY . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Appendix A . Task Force Membership . . . . . . . . . . . . . . . 41 Appendix B . Task Force Terms of Reference . . . . . . . . . . . 43 Appendix C . L i s t of P a r t i c i p a t i n g Ind iv idua l s , Companies

. . . . . . . . . . . . . . . . and I n s t i t u t e s . . 45 Appendix D . National Cancer I n s t i t u t e Data . . . . . . . . . . 4.7 Appendix E . I J C Great Lakes Research Advisory Board Terms

o f R e f e r e n c e . . . . . . . . . . . . . . . . . . . 55 Appendix F . I J C Great Lakes Research Advisory Board

Membership L i s t . . . . . . . . . . . . . . . . . . 57

Since World War 11, synthesized de te rgen t s have l a r g e l y replaced soap a s c l eans ing a i d s f o r laundering. Phosphates were t h e major de t e rgen t bui ld ing b locks u n t i l concern developed about t h e impact of t h e phosphate o n a l g a l growth i n t h e l a k e s and streams of North America. A s p re s su re grew t o r e s t r i c t t h e use of phosphate i n de t e rgen t s , Sodium N i t r i l o t r i a c e t a t e (NTA) became t h e most l i k e l y replacement. Because of f e a r t h a t NTA might cause cancer o r i n h e r i t a b l e changes i n man, t h e Surgeon General of t h e United S t a t e s Publ ic Health Serv ice i n 1970 requested t h a t NTA be withheld from use i n de te rgen t s i n t h e United S t a t e s . The Canadian Government d id not agree wi th t h e s i g n i f i c a n c e of t h e poss ib l e r i s k and has continued t o al low t h e use of NTA i n de te rgen t s i n Canada.

An I n t e r n a t i o n a l J o i n t Commission Great Lakes Research Advisory Board Task Force, s e l e c t e d from members of t h e North American s c i e n t i f i c community has reviewed t h e d a t a on which t h e o r i g i n a l U.S. dec i s ion was based. The Task Force has a l s o reviewed t h e l abora to ry s t u d i e s which have become a v a i l - a b l e s i n c e t h a t dec i s ion and has analyzed t h e Canadian experience wi th t h e use of NTA. This r e p o r t of t h e i r review i s t h e most up-tq-date a n a l y s i s of t h e h e a l t h impl ica t ions of t h e use of NTA. A second t a s k fo rce , i n i t i a t e d by t h i s Board t o s tudy t h e environmental impact of non-phosphate de tergent b u i l d e r s , has reported i t s in t e r im f ind ings on NTA t o t h e Board and these a r e recorded i n t h e c u r r e n t Annual Board Report t o t h e I J C . A complete r e p o r t of t h a t Task Force s tudy w i l l be published a t a l a t e r da t e .

In summary, NTA, though r e a d i l y absorbed by t h e r a t , mouse and dog, i s poorly absorbed from t h e gut by monkey o r man. The admin i s t r a t ion of huge doses i n r a t s and mice r e s u l t s i n increased e l imina t ion of z inc and an i n c r e a s e i n tumors of t h e kidney and/or u r ina ry t r a c t and l i v e r . Although t h e l a r g e s t doses r e s u l t e d i n an inc rease i n c e r t a i n s p e c i f i c tumors, t h e o v e r a l l incidence of tumors i n t h e t r e a t e d animals and t h e c o n t r o l animals d id not d i f f e r s i g n i f i c a n t l y .

While i t has been demonstrated t h a t NTA is indeed carc inogenic , t h e combination of t h e high dosage requi red t o i n i t i a t e t h e e f f e c t i n t h e t e s t animals , a long wi th t h e poor absorpt ion of NTA i n humans and t h e low l e v e l environmental incidence of NTA demonstrated i n t h e Canadian Monitoring Program would minimize t h e hazard t h a t NTA would have on human h e a l t h .

Because of t h e proven carc inogenic e f f e c t and i n s p i t e of t h e low l e v e l of hazard which NTA r ep resen t s , n e i t h e r t h e Task Force nor t h e Research Advisory Board f e l t capable of recommending a s p e c i f i c course of a c t i o n t o t h e I n t e r n a t i o n a l J o i n t Commission. The ques t ions posed i n t h e 1970 Woods' Committee r epor t have been answered, and a conserva t ive h e a l t h r i s k evalua- t i o n i s provided f o r cons ide ra t ion by the appropr i a t e j u r i s d i c t i o n s . It i s our f e e l i n g t h a t t h e dec i s ion on t h e use of NTA must be based on s o c i e t a l i n t e r e s t s r a t h e r than depending s o l e l y on s c i e n t i f i c cons ide ra t ions .

Great Lakes Research Advisory Board

The Task Force is indebted to W. R. Grace and Company, Monsanto Industrial: ' Chemicals Company and in particular to the Procter and Gamble Company for their assistance in assembling the many scientific references. We also wish to express our appreciation to the National Cancer Institute (NCI), particularly to Dr. R. A. Squire and Dr. G. Flamm and to Dr. I. A. Mitchell of U.S. Department of Health, Education and Welfare (H.E.W.) for their cooperation in making material available to the Task Force.

Finally, we express our thanks to the staff of Dr. Falk at the National Institute for Environmental Health Services (N.I.E.H.S.) and to the staff at the Great Lakes Regional Office of the International Joint Commission for their assistance in the preparation of this report.

In Canada, on August 1, 1970, r e g u l a t i o n s were put i n t o e f f e c t l i m i t i n g t h e P 0 content of de t e rgen t s t o 20%. A t t h a t t ime, Sodium N i t r i l o t r i a c e t a t e ( N T A ) ~ W ~ S considered by many a most l i k e l y s u b s t i t u t e , but l i t t l e was known about i t o t h e r than i t s chemical p r o p e r t i e s a s a b u i l d e r . A s h o r t t ime ' , l a t e r , t h e major de t e rgen t manufacturers i n t h e United S t a t e s , i n response t o concerns expressed by t h e Surgeon General, v o l u n t a r i l y d iscont inued t h e use of NTA pending f u r t h e r s tudy. I n Canada, NTA l e v e l s i n de t e rgen t s now average 15%.

. . . . . , The Research Advisory Board,. recognizzng t h e need f o r a r e - e ~ a l ~ a t i p * . of

NTA, appointed a Task Force t o review a l l e x i s t i n g information ;n<report ';oh';'' . ' . . . -

t h e h e a l t h impl i ca t ions of NTA. , . . .

The committee accepted t h e conclusions expressed by "The Woods Report1',* and reviewed and accepted t h e a d d i t i o n a l s t u d i e s on p o t e n t i a l mutagenic e f f e c t s of NTA which a r e e s s e n t i a l l y negat ive . The committee reviewed pre- pub l i ca t ion m a t e r i a l from two animal feeding s t u d i e s by N C I , one by NIEHS and t h e Progress Report on t h e Canadian Monitoring Program.

'NTA i s r e a d i l y absorbed by t h e r a t , mouse and dog but poorly :absorbed by t h e monkey and man ( see Table 1 ) . 72 hours a f t e r dos ing , l e s s than 3% remains i n any spec ie s s tudied . A small f r a c t i o n of t h e NTA which i s absorbed may be depos i ted on the ' sur face of t h e bone. No evidence of b io t ransformat ion of NTA i n mammals e x i s t s . NTA d id not show any compound-related e f f e c t s of reproduct ion, f e t a l s u r v i v a l , nor neonata l v i a b i l i t y . Fur ther , i t has been demonstrated t h a t NTA has no t e ra togen ic nor fe to- toxic p o t e n t i a l e i t h e r a lone o r i n combination wi th t h e heavy meta ls .

The acu te , subacute and chronic t o x i c i t y d a t a a r e summarized i n Tables 2 and 3. NTA s i n g l e dose a c u t e and subacute t o x i c i t i e s a r e very low and compatible t o t h a t of Sodium Tripolyphosphate (STP) which i t would r ep lace i n de te rgen t formulat ions.

Three new ca rc inogen ic i ty s t u d i e s a r e reviewed i n d e t a i l . I n t h e s t u d i e s , sodium NTA was fed t o r a t s a t doses from 200 t o 20,000 ppm i n d i e t and sodium NTA and a c i d form NTA t o r a t s and mice a t 5,000, 7,500 and 15,000 ppm i n d i e t . One study fed a c i d NTA a t 1,000 ppm i n ' d r i n k i n g water t o r a t s .

A s t a t i s t i c a l l y s i g n i f i c a n t e f f e c t was observed f o r tumors of t h e kidney and/or u r ina ry t r a c t i n a l l s t u d i e s . The d a t a a r e summarized i n Tables 2, 3 and 4, and t h e pre l iminary r e s u l t s w i l l be found i n Appendix D. There was a l s o a n observed i n c r e a s e i n l i v e r cancer al though t h e occurrence was only sporadic.

*See Page 40 f o r Reference

NTA may s tand by i t s e l f i n being absorbed r e a d i l y i n some s p e c i e s , no t metabol ized, bu t r e a d i l y el iminated by t h e kidney and ha rd ly s t o r e d wi th in t h e body.

The ten-fold i n c r e a s e i n u r ina ry e l imina t ion of z inc recorded i n a l l experiments admin i s t e r ing l a r g e doses of NTA must be considered s i g n i f i c a n t bu t i t s e t i o l o g i c s i g n i f i c a n c e towards t h e product ion of tumors of t h e u r i n a r y system has no t been e s t a b l i s h e d nor i t s mechanism e luc ida t ed . One can only s p e c u l a t e t h a t a s e n s i t i v e ba lance has been upse t by t h e l o s s of z i n c without similar l o s s e s of cadmium and o the r t r a c e elements. It i s concluded t h a t t h e p r i n c i p a l human exposure t o NTA is by d r ink ing water . The Progress Report on t h e Canadian Monitoring Program (now being prepared f o r r e l e a s e ) p u t s pub l i c water supply l e v e l s a t 10 ppb wi th a n occas iona l maximum of 50 ppb; 96% of a l l samples contained l e s s than 25 ppb.

The assessment of carc inogenic r i s k t o humans from p resen t l e v e l s of NTA i n t h e environment i s extremely tenuous. The e x t r a p o l a t i o n from high dose i n animals t o low dose i n humans must be done on a most conse rva t ive b a s i s . Table 5 t a b u l a t e s t h e maximum r i s k of u r i n a r y t r a c t tumor formation i n roden t s f o r each of t h e t h r e e s t u d i e s reviewed u t i l i z i n g t h e so-cal led l i n e a r s t r a i g h t l i n e a r i t h m e t i c method. To a s s e s s t h e p o t e n t i a l r i s k t o man, t h e d i f f e r e n c e i n abso rp t ion of NTA i n man and roden t s must a l s o be considered.

CONCLUSIONS

1. A comprehensive review of pas t r e p o r t s and more r ecen t d a t a on NTA and i t s s a l t s r e v e a l s no evidence of t e r a togen ic nor mutagenic p o t e n t i a l .

2. The Committee concluded t h a t t h e only concern under condi t ions of exposure t o NTA of t h e populat ion a t l a r g e , o r of occupat ional ly exposed persons, stems from f ind ings of carc inogenes is of t h e u r ina ry t r a c t of r a t s and mice given l a r g e doses of NTA over t h e i r l i f e t i m e s .

3. S t a t i s t i c a l a n a l y s i s of t h e animal d a t a was coupled wi th a c a l c u l a t i o n of t h e est imated maximum p r o b a b i l i t y of tumor formation i n rodents a t t h e l e v e l s of NTA t o which humans a r e exposed. The e s t ima tes of maximum r i s k f o r t h e environmental doses c i t e d i n Table 5 a r e gene ra l ly w i t h i n a n order of magnitude of 1 i n 2 mi l l i on .

4. I n o rde r t o a p p r e c i a t e t h e poss ib l e r e l a t i o n s h i p s of t hese e s t ima tes t o man, t h e fol lowing f a c t s should be taken i n t o account: (a ) The rodent p r o b a b i l i t i e s . incorpora te a l a r g e measure of conserva-

t i v e exaggerat ion. (b) Absorption of NTA from food o r water by human s u b j e c t s i s approxi-

mately one - f i f th of t h e degree of abso rp t ion demonstrable i n rats.

A t t h e r eques t of t h e Governments of t h e United S t a t e s and Canada, t h e I n t e r n a t i o n a l J o i n t Commission i n 1969 r epo r t ed on t h e causes and ex t en t of p o l l u t i o n t o t h e Great Lakes. The recommended remedia l measures included:

1. Reduction of t h e phosphorus conten t of d e t e r g e n t s , t h e amount of de te r - ' gen t used and replacement of phosphorus compounds i n d e t e r g e n t s by less harmful subs tances a s soon a s p o s s i b l e bu t no t l a t e r than 1972.

2. The inco rpo ra t i on of phosphate removal p rocesses i n municipal and . .

i n d u s t r i a l wastewater t rea tment p l a n t s .

Concern was expressed i n both c o u n t r i e s over t h e p o t e n t i a l h e a l t h and environmental e f f e c t s which might fo l l ow t h e s u b s t i t u t i o n - o f a l t e r n a t e subs tances f o r phosphorus i n d e t e r g e n t s . Sodium N i t r i l o t r i a c e t a t e (NTA) by v i r t u e of i t s chemical p r o p e r t i e s a s a b u i l d e r and c o s t was considered by many a most l i k e l y s u b s t i t u t e but l i t t l e w a s known about i t s o t h e r proper- t i e s . By 1970, s u f f i c i e n t NTA was being used i n bo th c o u n t r i e s t h a t s t u d i e s were implemented t o examine d r ink ing , r i v e r and l a k e water f o r NTA.

I n Canada on August 1, 1970, r e g u l a t i o n s were put i n t o e f f e c t l i m i t i n g t h e P 0 con ten t of d e t e r g e n t p roducts t o 20%. The fo l lowing year t h e 2 5 Canadlan Nat iona l NTA Monitoring Program was e s t a b l i s h e d t o a c q u i r e d a t a upon which a p p r o p r i a t e conc lus ions could be based. A Progress Report on t h i s Program cover ing t h e per iod t o September 1975 is imminent.

I n December 1970, i n response t o concerns expressed by t h e Surgeon General of t h e United S t a t e s Pub l i c Heal th Se rv i ce , t h e major d e t e r g e n t manufacturers i n t h e U.S. v o l u n t a r i l y d i scont inued t h e u s e of NTA pending f u r t h e r s tudy and eva lua t ion of t h e p o t e n t i a l hazard t o human h e a l t h . A s a r e s u l t of t h i s a c t i o n , NTA consumption i n t h e United S t a t e s has dropped t o a n es t imated 10 m i l l i o n pounds annual ly .

I n January 1975, t h e Canadian Government a f t e r due c o n s i d e r a t i o n , f u r t h e r reduced t h e maximum P205 l e v e l s t o 5%. A l a r g e p a r t of t h e phosphorus r educ t ion has been made up through NTA s u b s t i t u t i o n . I n e a r l y 1972, indus t ry- wide NTA conten t i n d e t e r g e n t s averaged 6% and i n 1975, 250 m i l l i o n pounds of d e t e r g e n t s were purchased wi th an average NTA conten t of 15%. It h a s been es t imated t h a t t h e c u r r e n t annual consumption of NTA i n Canada is 60 m i l l i o n pounds.

The 1975 Annual Report of t h e Water Qua l i t y Board t o t h e I n t e r n a t i o n a l J o i n t Commission (IJC) recommended t h a t a l l Great Lakes. S t a t e s should impose

phosphorus l i m i t a t i o n s on d e t e r g e n t s s o l d i n t h e bas in . Canadian po l i cy does no t p r o h i b i t t h e u se of NTA a s a replacement f o r phosphorus whi le i n t h e United S t a t e s t h e Surgeon Genera l ' s r eques t of 1970 is s t i l l being observed. I n t h e meantime, r e sea rch has been completed which appears t o answer t h e ques t ions r a i s e d by "The A s s i s t a n t S e c r e t a r y ' s Ad Hoc Group on NTA, 1972" (The Woods Report) w i th regard t o poss ib l e mutagenic e f f e c t s . Also, t h e Nat iona l Cancer I n s t i t u t e has j u s t r e l e a s e d d e t a i l s on two a d d i t i o n a l animal feeding s t u d i e s bear ing on t h e ques t ion of NTA ca rc inogen ic i ty . F i n a l l y , . t h e Progress Report on t h e Canadian Monitoring Program is c u r r e n t l y being r e l eased .

The Research Advisory Board (RAB) recognized t h a t i t w a s now a p p r o p r i a t e t o re-examine t h e h e a l t h e f f e c t s of NTA and appoin ted , a s is t h e po l i cy of t h e RAB, one-of i ts members, Paul Foley - Chairman of "The Task Force on t h e ~ e a l t h Imp l i ca t ions of NTA" and reques ted t h a t he nominate s c i e n t i s t s repre- s e n t i n g d i s c i p l i n e s p e r t i n e n t t o t h e s u b j e c t t o c o n s t i t u t e t h e t a s k fo rce . The RAB appointed t h e f i v e s c i e n t i s t s l i s t e d i n Appendix A t o s e rve on t h e t a s k force . Also l i s t e d are t h e two t e c h n i c a l adv i so r s . The terms of r e f e r e n c e of t h e t a s k f o r c e a r e given i n Appendix B.

The t a s k f o r c e f i r s t m e t - i n September and arranged subsequent meetings t o provide a n oppor tun i ty t o meet wi th c o n t r i b u t i n g s c i e n t i s t s , review r e p o r t s and o t h e r a v a i l a b l e d a t a and t o develop t h i s r e p o r t . A l i s t of persons , companies and i n s t i t u t e s who m e t w i th t h e t a s k f o r c e i s given i n Appendix C. The Committee reviewed and i n gene ra l accepted t h e conclus ions expressed by "The Woods Report". The r e p o r t on "Current S t a t u s of t h e Environmental and Human Sa fe ty Aspects of N i t r i l o t r i a c e t i c Acid (NTA)" by Arthur D. L i t t l e , I nc . , ha s served as a u s e f u l r e sou rce t o c u r r e n t s c i e n t i f i c l i t e r a t u r e .

This r e p o r t d i s c u s s e s t h e tox ico logy of NTA and w i l l encompass t h e fo l lowing a r e a s :

1. E x i s t i n g r e p o r t s 2 . Pharmacology 3. .Reproduct ion and f e t o t o x i c i t y 4. Mutagenici ty 5. Acute, subacute and chronic t o x i c i t y 6. Carc inogenic i ty 7. ' Exposure l e v e l s

EXISTING REPORTS

With regard t o human h e a l t h e f f e c t s , two a r e a s of concern were i d e n t i f i e d by t h e Woods r e p o r t . For one, i t w a s noted t h a t " the re a r e no r e p o r t s of muta t iona l s t u d i e s on p o s s i b l e NTA metabo l i t e s o r on d e r i v a t i v e s t h a t might be formed i n t h e environment p r i o r t o human exposure. While t h e metabolism of NTA i n mammals appears i n s i g n i f i c a n t , a t t empt s should be cont inued t o i d e n t i f y p o t e n t i a l me tabo l i t e s , and t h e mutagenici ty of t h e s e subs tances should be t e s t e d thoroughly". Secondly, i t w a s t h e opin ion of t h e Woods Committee t h a t " the experimental d a t a a v a i l a b l e a t t h i s t ime d i d no t permit a conclusion as t o whether NTA is o r i s n o t carcinogenic".

Since t h e appearance of t h e Woods r e p o r t , a d d i t i o n a l s t u d i e s r e g a r d i n g t h e s e two concerns have been completed. S t u d i e s . . , . on p o t e n t i a l mutagenic ., . e f f e c t s of NTA a r e e s s e n t i a l l y nega t i ve .

Also, t h e Na t i ona l Cancer I n s t i t u t e h a s j u s t r e l e a s e d d e t a i l s on two a d d i t i o n a l animal f e ed ing s t u d i e s b e a r i n g on t h e q u e s t i o n of t h e ca r c ino - g e n i c i t y o f NTA. A p r e l im ina ry r e p o r t o f r e s u l t s of a n NIEHS s tudy is a l s o a v a i l a b l e . These documents w i l l b e eva lua t ed i n t h i s r e p o r t . F i n a l l y , t h e P rog re s s Report on t h e Canadian Monitor ing Program, though s t i l l n o t r e l e a s e d , was a v a i l a b l e t o t h e Committee f o r review.

PHARMACOLOGY . , . , . ,. . . , . . . .

There i s c o n s i d e r a b l e v a r i a t i o n i n t h e deg ree of a b s o r p t i o n of NTA from t h e g a s t r o i n t e s t i n a l t r a c t among t h e d i f f e r e n t s p e c i e s t e s t e d (Table 1 ) . It i s r e a d i l y absorbed by t h e r a t , mouse and dog and poo r ly absorbed by t h e monkey and man. I n t h e ra t t h e pr imary r o u t e of e l i m i n a t i o n is t h e u r i n a r y t r a c t , wh i l e i n man i t i s t h e i n t e s t i n a l t r a c t . I n man, approximately:75% of a s i n g l e o r a l dose i s recovered i n f e c e s a f t e r 72 hou r s , wh i l e 12% i s found i n t h e u r i n e . A f t e r 72 hours i t i s e s t ima t ed t h a t less t h a n 3% remains i n any s p e c i e s s t u d i e d .

Two s t u d i e s were c a r r i e d ou t i n human v o l u n t e e r s , f ou r v o l u n t e e r s i n t h e f i r s t s t u d y r e c e i v i n g 1 mg ~ T A l k g body weight dose ; t h e o t h e r g i v i n g each s u b j e c t a t o t a l dose of 1 0 mg of C ' ~ - N T A i n a g e l a t i n c apsu l e . I n t h e f i r s t s t u d y 72 t o 98% o f t h e t o t a l dose was recovered , w i t h 5 t o 12% i n t h e u r i n e , t h e rest i n t h e f e c e s ; w h i l e i n t h e r a d i o a c t i v e s t u d y t h e r e s u l t s of e i g h t i n d i v i d u a l exper iments were pooled g i v i n g a 12.3 + 7.4% u r i n a r y r ecove ry and a t o t a l r ecovery of 89.3 + 12.4%. Human and monkey r ecove ry d a t a sugges t t h a t p r imates have low a b s o r p t i o n of NTA, r a b b i t s a somewhat h i g h e r absorp- t i o n rate and rats, mice and dogs much h i g h e r a b s o r p t i o n rates (Table 1 ) .

Evidence ob t a ined from r a d i o i s o t o p e t r a c e r t e chn iques and mass s p e c t r o - g r a p h i c a n a l y s i s i n d i c a t e s t h a t t h e r e is no s i g n i f i c a n t b io t r ans fo rma t ion of NTA i n mammals.

A s m a l l f r a c t i o n of t h e NTA which is absorbed may be d e p o s i t e d on t h e s u r f a c e of bone, where i t h a s a l ong h a l f - l i f e . D e t e c t a b l e amounts (300 ~.lg/g) of NTA were found i n ra t bone 30 days a f t e r c e s s a t i o n of f i v e 1 0 mg/kg dose s of NTA. Depos i t i on i n bone r e a c h e s a p l a t e a u . S t u d i e s i n t h e ra t have i n d i c a t e d t h a t t h e p r e sence of NTA a f f e c t s n e i t h e r bone s t r u c t u r e no r s t r e n g t h .

REPRODUCTION AND FETOTOXICITY

A two-generat ion s t u d y of d i e t a r y NTA a t l e v e l s up t o 5,000 ppm i n rats d i d n o t show any compound-related e f f e c t s on r e p r o d u c t i o n , f e t a l s u r v i v a l , n o r neona t a l v i a b i l i t y .

TABLE 1

MAMMALIAN ABSORPTION AND EXCRETION

OF NTA

Oral Dose of NTA Percent of Dose (48-72 hour) Excreted)

Species* mg/kg P C ~ /kg Urine Feces

R a t

Dog

Rabbit

Monkey

Mouse

Man

*The numbers of animals per spec ie s were: rat ( 3 ) , dog ( I ) , r a b b i t ( I ) , monkey ( I ) , mouse ( 5 ) , and man (8 ) . Oral doses were given a f t e r a 10-16 hr . f a s t (mice non-fasted) and water -- ad l i b . post-dosing. Radioact ive doses a r e c a l c u l a t e d from o r i g i n a l papers and a r e only approximate except f o r mice. Radioac t ive NTA w a s given as t h e disodium salt except f o r mice where f r e e a c i d w a s given i n phosphate b u f f e r pH=7.4

The t e ra togen ic p o t e n t i a l of NTA has been t e s t e d i n a number of spec ie s . NTA administered t o r a t s , mice, o r r a b b i t s throughout t h e period of major organogenesis a t doses a s high a s 500 mg/kg/day i n r a t s and mice and 250 mg/kg/day i n r a b b i t s d id not demonstrate any t e ra togen ic o r o t h e r Eetotoxic e f f e c t .

The p o s s i b i l i t y of f e t o t o x i c e f f e c t s a s a r e s u l t of p o t e n t i a t i o n of heavy metal e f f e c t s by NTA-metal c h e l a t e s has a l s o been inves t iga t ed . S tudies i n t h e r a t have not shown increased f e t o t o x i c i t y when NTA and cadmium o r mercury were administered s imultaneously.

It may be concluded t h a t NTA does not have any f e t o t o x i c p o t e n t i a l , e i t h e r a lone o r i n conjunct ion wi th heavy metal 's , when administered experimental ly t o pregnant animals.

MUTAGENICITY

The Woods Committee concluded t h a t f u r t h e r s t u d i e s were necessary t o a s s u r e l a c k of mutagenic r i s k wi th NTA. Thei r conclusions were based upon experiments done up t o 1972. A t t h a t t ime, a number of mutagenici ty t e s t s i n a v a r i e t y of systems had e i t h e r been accomplished o r were ongoing. P la te . t e s t s with SaZmoneZZa t y p h i w i w n had been done and no mufagenic tendency of NTA was noted. Tes t s with E. c o z i i n c u l t u r e medium a l s o ind ica t ed no mutagenic p o t e n t i a l of NTA and f u r t h e r showed no p o t e n t i a t i o n of t h e e f f e c t s of known mutagens ( e t h y l methanesulfonate and x-ray). Tes t s i n DrosophiZa were a l s o negat ive . I n mammals, a s e r i e s of dominant l e t h a l t e s t s i n mice and r a t s was performed and were negat ive . I n a host-mediated t e s t i n mice which u t i l i z e d S. typhimwliwn t h e r e was no evidence of any i n c r e a s e i n mutat ions. The Woods Committee a l s o repor ted on cytogenet ic s t u d i e s us ing c e l l l i n e s from Kangaroo r a t s ; one which was nega t ive , and one which r e s u l t e d i n a low l e v e l of chromatid breaks a t 500 mg/l. Marrow c e l l s from r a t s exposed t o NTA a t a dose of 250 mg/kg given i n t r a p e r i t o n e a l l y , showed no e f f e c t s . Since t h a t t ime a s u b s t a n t i a l number of a d d i t i o n a l s t u d i e s on a v a r i e t y of systems have been performed. S tudies on Neurospora have been negat ive . Chromatid a b e r r a t i o n s have been shown i n t e s t s us ing cu l tu red human lymphocytes wi th s u b l e t h a l concen t ra t ions of Na3NTA. No chromatid a b e r r a t i o n s were found, however, i n r ecen t -- i n vivo t e s t s us ing rodent bone marrow c e l l s . Dominant l e t h a l t e s t s i n DrosophiZa have proven t o be nega- t i v e . Dominant l e t h a l t e s t s i n mice given 1,000 ppm NTA i n d r ink ing water have a l s o shown no mutagenic e f f e c t s . Her i t ab le t r a n s l o c a t i o n t e s t s i n mice r ece iv ing a similar dose have l i kewise shown no p o s i t i v e e f f e c t s . Attempts t o i d e n t i f y pos tu la ted me tabo l i t e s have a l l been unsuccessful and the re fo re t h e i r t e s t i n g wi th hypo the t i ca l compounds f o r mutagenic p o t e n t i a l is i r r e l - evant .

The Task Force concludes t h a t t h e NTA d a t a i n d i c a t e no p o t e n t i a l f o r t h e induct ion of gene mutat ions o r t ransmi t ted gene t i c e f f e c t s . The only p o s i t i v e e f f e c t s have been chromosome breakage -- i n v i t r o a t high dose l e v e l s ( 1 0 2 ~ ) . The d a t a conta in no evidence t h a t NTA would present a mutagenic hazard i n t h e environment.

ACUTE, SUBACUTE AND CHRONIC TOXICITY

The acu te , subacute and chronic t o x i c i t y has been reviewed by t h e Woods Committee a s wel l a s by Thayer and Kensler. The d a t a a r e summarized i n Tables 2 and 3. Stud ies on s i n g l e dose acu te and subacute exposure t o NTA i n d i c a t e t h a t its l e v e l of t o x i c i t y is very low and comparable wi th t h a t of sodium t r ipolyphosphate (STP), which i t would r ep lace i n de te rgen t formula- t ions.

DEFINITION

A neoplasm has been def ined by Ewing a s an uncont ro l led new growth of t i s s u e . The d i s t i n c t i o n between benign and malignant neoplasms i s not always p e r f e c t l y c l e a r ; however, s e v e r a l of t h e fol lowing c h a r a c t e r i s t i c s d i s t i n g u i s h a malignant from a benign neoplasm: a h igh m i t o t i c r a t e and t h e occurrence of abnormal mitoses, ,abnormal c e l l s i z e s and hyperchromatic n u c l e i , l o s s of s t r u c t u r a l arrangement of t h e - c e l l s , bu t most important ly t h e invas ive c h a r a c t e r i s t i c and a b i l i t y t o me tas t a s i ze d i s t i n g u i s h malignant from benign and normal t i s s u e .

I n experimental s t u d i e s f o r purposes of t e s t i n g t h e ca rc inogen ic i ty of va r ious agents , it is customary no t t o s e p a r a t e benign and malignant tumours, s i n c e t h e h i s t o l o g i c a l c h a r a c t e r i s t i c s a r e n o t always r e a d i l y i n t e r p r e t a b l e . Secondly, benign tumours may at some l a t e r t ime behave l i k e malignant ones; hence s t a t i s t i c a l cons ide ra t ions i n t h i s r e p o r t cover both c a t e g o r i e s .

EXPERIMENTAL DESIGN

I n t h e Stanford Research I n s t i t u t e (SRI) des ign t h e h i g h e s t dose which d i d not cause any dea th among t h e animals fed e i g h t weeks on t h e d i e t was 20,000 ppm N a NTA.H 0, which w a s chosen even though a 10% weight decrement 2 w a s observed qn males and a 20% decrement i n t h e females. The lower doses were 2,000 and 200 ppm N a NTA.H20. A c o n t r o l group w a s kept on t h e same d i e t without t h e de te rgen t b u d d e r . Twenty-four animals of each sex were chosen f o r each l e v e l and t h e experiments were s t a r t e d when t h e animals were seven t o e i g h t weeks old. Feeding of t h e s e d i e t s was continued f o r two years .

I n t h e L i t t o n B ione t i c s s t u d i e s t h e dose l e v e l s were s i m i l a r l y determined f o r both sexes of t h e two spec ie s us ing t h e f r e e a c i d and t h e sodium sa1.t of NTA. No l e t h a l i t y w a s observed wi th t h e h ighes t dose l e v e l s chosen on e ight - week feeding s tud ie s . For rats and mice 15,000 ppm of t h e a c i d w a s t h e h i g h e s t d o s e chosen; a second l e v e l chosen w a s h a l f t h a t dose. The sodium salt w a s given t o rats a t t h e same doses, but mice rece ived only 5,000 ppm and one-half t h a t dose i n t h e i r d i e t . F i f t y animals of each sex were s t a r t e d on t h e d i e t when s i x weeks of age and f ed these d i e t s w i th de te rgen t b u i l d e r o r t h e c o n t r o l d i e t f o r 18months , but observed f o r 24 months.

TABLE 2

SUMMARY OF TOXICITY ON ORAL INTAKE OF NTA

Compound and Dose Spec ies (glkg b.w.

Acute Tox ic i ty

Mor t a l i t y Pathology Source

Acid

Na s a l t

SD* r a t s 3/10 mala ise , l oose s t o o l s , anorexia Rub i n 11 groups of 10

lower 11

0.5 - 3.5 r a b b i t s 7 groups of 4

none observed

SD* r a t s groups of 10 0

- - - -

no s i g n i f i c a n t pathology

Rhesus decreased motor a c t i v i t y , p a r a l y s i s , 11

groups of LD500.75 dea th , hemorrhagic a r e a s of stomach, 1, 2, and 3 vomiting i n one minute

Dogs LDS05. 0 vomiting, recovery groups of 4

TABLE 3

SUMMARY OF TOXICITY ON ORAL INTAKE OF NTA

Sub-Acute Tox ic i ty -. .

- Compound and Dose Species Pathology o r Biochemical Abnormali t ies Source

i n D ie t

Acid up t o 5% SD* r a t s A t 5%: l o s s of weight , anorexia , M o r t a l i t y 318 Rubin f o r 3 mo. 4 groups No abnorma l i t i e s i n kidney, lung, G I and GU t r a c t ,

of 8 sp leen , h e a r t , b r a i n , nervous t i s s u e and l i v e r

Na s a l t 2%, 0.2% f o r 3mo .

Na s a l t 2% f o r 1 mo.

Na s a l t ' . 2%, 0.5% 0.02% f o r 3 mo.

SD* r a t s weanlings 5 groups of 20

CR** r a t s 10 female 14 male

r a t s

A t 2%: en larged k idneys , decrease i n weight ga in , Nixon hydronephrosis i n 9/10 males, 319 females A t 0.2% no e f f e c t

A t 1%: e x t e n s i v e hydropic degenera t ion of t u b u l e s A t 0.75%: mild degenera t ion of t ubu le c e l l s i n 4/10 males

A t 2%: u r ina ry l e v e l of Zn, Ca, Na inc reased A t 0.2%: none observed

Depressed growth r a t e s , u r ina ry pH inc reased , Zn, Ca, Na' increased

Michael 6 Wakim

2%: Local ized kidney l e s i o n s a f t e r 3 mo. EM s tudy , Niewenhuis membrane l i m i t e d i n t r a c e l l u l a r vacuole formation

Acid 2%, 0.5% r a t s 2%: t u b u l e d i l a t a t i o n , hydropic degenera t ion of tubule A . D. L i t t l e 0.03% c e l l s , l o c a l i z e d l e s i o n s , a t 3 mo. Some changes a l ready f o r 3 mo. d e t e c t a b l e a t 40 and 65 days.

No changes observed a t lower l e v e l s

Na S a l t 1% i n H20 SD* r a t s Hydropic degenera t ion of t ubu le s , uremia, dea th 1 mo.

Goyer

*Sprague-Dawley **Charles River

Protocols f o r handling and maintaining t h e animals were made a v a i l a b l e t o t h e committee a s were t h e d e t a i l s of autopsy a n d pathology pro tocols . The s tudy focused mainly on t h e d e t e c t i o n of neoplas ia i n these animals .

I n a s tudy c a r r i e d out a t NIEHS approximately 200 Charles River white male r a t s were given 0.1% (1,000 ppm) tr isodium n i t r i l o t r i a c e t i c a c i d i n d r ink ing water. Animals were s a c r i f i c e d when moribund o r when tumors were ' g r o s s l y ev ident . The experiment was terminated a f t e r two yea r s and included a s i m i l a r number of c o n t r o l animals. The r e s u l t s of t h i s experiment a r e being prepared f o r pub l i ca t ion and were made a v a i l a b l e t o t h e Committee i n t h e form of a prel iminary d r a f t .

(a ) Weight Gain

A s could be expected male and female animals on t h e h ighes t dose l e v e l s (SRI) s tayed below t h e o t h e r dose l e v e l and c o n t r o l s i n weight gain. I n t h e L i t t o n Bionet ics s tudy where t h e feeding of t h e "detergent d i e t " was discontinued a f t e r 18 months a s t r i k i n g i n c r e a s e i n weight from t h e r e on was observed i n t h e h ighes t dose groups.

(b) Mor ta l i t y

The s u r v i v a l of t h e animals i n a l l groups was comparable except f o r t h e male r a t s on t h e h ighes t dose i n t h e SRI s tudy which showed e a r l i e r dea th -- 75% had d ied be fo re t h e te rminat ion of t h e experiment a t 104 weeks; most of t hese dea ths occurred dur ing the second year of t h a t s tudy and were uniformly spread through t h a t time i n t e r v a l .

I n t h e NIEHS s tudy, t h e o v e r a l l m o r t a l i t y f o r NTA t r e a t e d a n d c o n t r o l r a t s d i d not d i f f e r . However, a higher propor t ion of t he t r e a t e d animals appeared t o d i e i n t h e f i r s t 550 days of t h e s tudy (19.7% vs. 11.2%, p c0.05).

(c ) Tumor Incidence

i. Renal tumors

To ta l tumor inc idence i n a l l s t u d i e s was s i m i l a r f o r a l l groups of r a t s due t o t h e high s u s c e p t i b i l i t y t o tumors of t h e reproduct ive system, bu t when t h e tumor inc idence was compared by organ systems i t became c l e a r t h a t a s t a t i s t i c a l l y s i g n i f i c a n t e f f e c t was observed f o r tumors of t h e kidney and u r i n a r y t r a c t . These tumors were observed i n r a t s of both sexes on t h e h ighes t dose i n t h e SRI s tudy us ing t h e sodium salt of NTA. With t h e a c i d females and males showed u r i n a r y t r a c t cancer bu t only t h e female r a t s showed u r i n a r y bladder cancer induct ion .

I n mice t h e a c i d produced neoplasms of t h e kidney i n both sexes, '

b u t s t a t i s t i c a l l y s i g n i f i c a n t d i f f e r e n c e s were observed only i n t h e males. The sodium s a l t produced no tumors of kidney, u r e t e r , o r

bladder , probably because of t h e low dose used a s h igher doses : c o u l d . n o t be t o l e r a t e d .

The r e l evan t information inc luding t h e s t a t i s t i c a l s i g n i f i c a n c e i s given i n Appendix D obtained from t h e pre l iminary N C I r e l e a s e .

The eva lua t ion of t hese d a t a regard ing human h e a l t h hazards r e q u i r e s e x t r a p o l a t i o n from high t o low dose l e v e l s and comparison of t h e

. . pharmacokinetics of NTA i n rodents and humans f o r which f o r t u n a t e l y some d a t a a r e ava i l ab le . I n a d d i t i o n , d a t a a r e a v a i l a b l e f o r some o t h e r de t e rgen t b u i l d e r s and c h e l a t i n g agents t o a l low some comparisons.

I n t h e NIEHS study t h e l a r g e s t t reatment-control d i f f e r e n c e i n tumor inc idence was a s soc ia t ed wi th r e n a l tumors (Table 4) . Although t h e t o t a l number of tumors observed i n NTA t r e a t e d r a t s exceeded c o n t r o l r a t s , t h i s d i f f e r e n c e was accounted f o r e n t i r e l y by t h e r e n a l tumors.

ii. Tumors of o t h e r organs

The s tudy discovered a l s o some o t h e r s t a t i s t i c a l l y s i g n i f i c a n t carc inogenic dose responses i n these animals; i . e . an increased inc idence of cancer of t h e l i v e r and of t h e lung i n female r a t s on t h e high NTA d i e t , which was no t observed i n t h e male animals.

. , . Although tumors, a p a r t from u r ina ry t r a c t tumors fol lowing NTA feeding, a r e of i n t e r e s t t h e occurrence i s sporadic i n s t u d i e s t o d a t e and only u r ina ry t r a c t tumors show a cons tant s t a t i s t i c a l l y v a l i d inc rease fol lowing NTA inges t ion .

Comparison wi th S tud ie s on Other Chela t ing Agents and Detergent Bui lders

Comparisons of t h e carcinogenic p o t e n t i a l of NTA with t h a t of o t h e r c h e l a t i n g agents and de te rgen t b u i l d e r s i s not poss ib l e because appro- p r i a t e s t u d i e s have not been made a v a i l a b l e t o t h e Task Force. However, t h e r e a r e s i m i l a r i t i e s i n t h e t o x i c e f f e c t s of NTA and o t h e r c h e l a t i n g agen t s , p a r t i c u l a r l y on t h e kidney.

Comparisons of o t h e r de t e rgen t b u i l d e r s and c h e l a t i n g agen t s i n lab- o ra to ry animals have shown t h a t abso rp t ion of h igh doses of de t e rgen t b u i l d e r s is poss ib l e on o r a l i n t a k e of co r ros ive de te rgen t b u i l d e r s which cause, bes ides t h e l o c a l damage, kidney t o x i c i t y ; i .e. t u b u l a r damage due t o carbonate and s i l i c a t e . I n s t u d i e s on o t h e r c h e l a t i n g agen t s such as EDTA hydropic degenera t ion of r e n a l t ubu les w a s a l s o observed when t h e c h e l a t i n g agent w a s given in t ravenous ly .

It should be noted however t h a t ready comparisons wi th o the r c h e l a t i n g agents may not be app l i cab le . EDTA is no t wel l absorbed from t h e g a s t r o i n t e s t i n a l t r a c t , and o t h e r s l i k e c i t r i c a c i d a r e absorbed and metabolized i n t h e body.

TABLE 4

INCIDENCE OF TOTAL TUMORS AND RENAL ADENOMAS I N RATS

GIVEN 0.1% (1000 ppm) NTA I N DRINKING WATER FOR UP TO TWO YEARS TUMORS/ANIMALS AT RISK, NIEHS STUDY - THE WOODS REPORT

* Inc ludes 4 r e n a l adenocarcinomas

Tumor Group

Renal Adenomas*

T o t a l Tumors

T o t a l Tumors Less Renal Adenomas

"P"

p (0.01

p <0.01

N.S.

Cont ro l

5/186

111/186 ,

-

Trea t ed

29/183

131/183

106/186 1 102/183

NTA may stand by itself in being absorbed readily in some species, not metabolized, but readily eliminated by the kidney and hardly stored. However, when chelating agents enter the circulation they will cause damage to the kidney, whether they are NTA, EDTA, polyphosphate, carbonate, or silicate.

Hypothesis of Carcinogenic Action of NTA

In order to assess the potential health hazard to man of very low dose levels of NTA it would be helpful to understand as much about the mechanism of action as possible. Unfortunately, little information is available to elucidate the mechanism of carcinogenesis. Two suggestions have been made. One deals with the formation of a nitrosamine by certain microorganisms in the sewage, the other with chelation of an essential element in the mammalian organism and its excretion.

Nitrosamine production

The production of N-nitrosoiminodiacetate (NIDA) from NTA and nitrate by mixed cultures of microorganisms has been reported. It required a mixed population of Pseudomonas and 30 days of acclimatization to obtain the nitrosamine. In cell-free extracts of Pseudomonas A and C it was possible to obtain the NIDA from nitrate and NTA in vitro. Studies on the mutagenicity of the compound were negative even after hepatic microsomes had been added. However, the compound showed some antimicrobial activity.

In rat and mouse carcinogenicity studies on NTA alone, NTA with nitrate and nitrite or the anticipated N-nitroso derivative (NIDA) were administered in the drinking water at high dose levels. No difference in tumor incidence'was seen even though the rats con- sumed a total of 40 g NTA or iminodiacetate, but the number of animals on study was too small for adequate statistics. The types of tumors expected in animals on nitrosamine treatment were not increased. In the mouse study the dose of 5 g NTA/liter or 2 g NIDA/liter did not significantly increase the number of lung tumors, the total number of lung tumors per animal, nor the number of tumor-bearing animals in this study. The slight increase in lung-adenoma-bearing animals and the small increase in adenomas per mouse in the group given NTA and sodium nitrite in water may be suggestive of a slight stimulation of lung adenoma production, but the inhibitory effect of NIDA at 2 g/liter on lung adenoma incidence may suggest that the system is not suitable for these studies.

(ii) Essential element chelation -- zinc

The.second hypothesis is based on chelation of an essential element. Several studies on NTA presented evidence that even on a medium high dose increased mobilization of zinc is detectable in urine. Observations have been made on rats given sodium NTA in the diet at levels of 1,500 or 5,000 ppm showing kidney pathology and elevated urinary zinc levels. Similar increases in other metals were not

seen. A s t h e r e s u l t of such mobili .zation, some z inc has a l s o been de tec ted i n bone i n these experiments.

Although a ten-fold i n c r e a s e i n u r i n a r y e l imina t ion of z inc has been recorded i n a l l experiments adminis te r ing l a r g e doses of NTA i t s e t i o l o g i c s i g n i f i c a n c e towards t h e product ion of tumors of t h e u r i n a r y system has not been e s t ab l i shed nor i t s mechanism e l u c i - dated. One can only specu la t e , however, without adequate b a s i s but by analogy, t h a t a s e n s i t i v e balance has been upset by l o s s of z inc without causing f o r i n s t ance a s i m i l a r dep le t ion of cadmium o r lead .

This balance is e s s e n t i a l t o prevent t h e t o x i c i t y of cadmium. Cadmium may be a carc inogenic agent f o r a kidney, a s was r e c e n t l y suggested i n epidemiologic s t u d i e s by Kolonel: such l eads need f u r t h e r study. I f so, t h e dep le t ion of z inc would have been a b l e t o b r ing out t h i s t o x i c i t y .

The l i t e r a t u r e r e p o r t s t h a t p a r e n t e r a l l y administered EDTA t r i g g e r s t h e e l iminat ion of z inc i n t h e u r i n e i n experimental animals a s we l l a s humans. These papers do not d e a l wi th long-range e f f e c t s of such t reatment a s i t is usua l ly of s h o r t du ra t ion and t h e p a t i e n t s a r e not followed f o r any l eng th of t ime but r e n a l t o x i c i t y has been repor ted a s wel l a s u r i n a r y e l imina t ion of l a r g e amounts of z inc . A connection between t h e two events has not been estab- l i s h e d except t h a t some enzymes have been shown t o l o s e t h e i r bound z inc and be i n a c t i v a t e d . This is p a r t i c u l a r l y t r u e f o r r e n a l a l k a l i n e phosphatase and hepa t i c glutamate dehydrogenase. The important enzyme carbonic anhydrase -- a l s o a z inc enzyme -- is not a f f ec t ed .

I f i t were t r u e t h a t dep le t ion of z inc from s p e c i f i c organs and an imbalance of s p e c i f i c t r a c e elements were e t i o l o g i c a l l y r e l a t e d t o carc inogenes is t h i s could se rve a s a b a s i s f o r experiments t o prove t h i s r e l a t i o n s h i p . If i t could be proven, i t would s e r v e t o con- s i d e r NTA a "secondary" carcinogen, i . e . an agent which only a t a c e r t a i n l e v e l produces a condi t ion which may l ead t o cancer development. Levels of NTA t h a t could no t adverse ly a f f e c t t h e balance of t r a c e elements would not be expected t o have an adverse e f f e c t i n r e l a t i o n t o carcinogenesis .

Humans w i l l be exposed t o NTA v i a percutaneous absorpt ion o r acc iden ta l i nges t ion of NTA-containing de te rgen t s , NTA r e s idues on d i shes a n d . u t e n s i l s , and NTA presen t i n water used f o r cooking and dr inking . For t h e purposes of a s ses s ing t h e exposure of t h e populat ion a t l a r g e , t h e l e v e l of NTA i n t h e workplace w i l l no t be included.

Human exposure t o NTA from a c c i d e n t a l i nges t ion i s . i m p o s s i b l e t o es t imate . Based on comparative pharmacokinetics and emetic a c t i v i t y of NTA-containing de te rgen t s , humans w i l l be a t minimal r i s k from a c c i d e n t a l i nges t ion . I n t h i s context t h e a c t i v i t y of NTA is equiva lent t o STP (6.0 mg/kg i n dogs) .

. .

The amount of NTA present a s a r e s idue on d i shes i s d i f f i c u l t t o calcu- l a t e . Surveys i n d i c a t e t h a t l e s s than 5% of t h e populat ion use laundry de , te rgents conta in ing an average of 15% NTA, f o r dishwashing. This p r a c t i c e r e p r e s e n t s t h e l a r g e s t source of NTA f o r t h e small number of peopie who use NTA-containing laundry de te rgen t f o r dishwashing and do not r i n s e t h e i r d i shes .

An examination of t h e chemistry of NTA i n d i c a t e s t h a t food chain t r ans - mission i s un l ike ly . No d a t a a r e a v a i l a b l e on NTA l e v e l s i n e d i b l e foods tu f f s (vegetables , f i s h , b i r d s , e t c . ) .

Previous r e p o r t s i n a s ses s ing t h e environmental l e v e l s of NTA concluded t h a t t he l e v e l s found i n dr inking water would r e s u l t i n an average d a i l y a d u l t i n t a k e of 63-100 pg.

Since then, t h e Progress Report on t h e Canadian Monitoring Program has been r e l eased . The NTA va lues i n publ ic water supp l i e s a r e r epor t ed t o be u s u a l l y l e s s than 10 p g l l (ppb) wi th a l e v e l of 50 p g / l (ppb) a s a n occas ional maximum., Some h igher va lues (up t o a few ppm) a r e repor ted i n shal low w e l l s which were contaminated by p o l l u t i o n from s e p t i c tank dra inage and r e a l l y u n f i t f o r a d r ink ing water source.

6 Under t h e present Canadian use p a t t e r n (15% NTA, . i . e . 60 x 10 l b s l y r )

d r ink ing water has been found t o con ta in an average of 5 p g / l w i th 96-% of a l l samples conta in ing l e s s than 25 p g l l . Only one c i t y a t one time had a l e v e l of 59 p g l l and t h i s c i t y now has a l e v e l approaching 5 pg / l . A t a consumption of 2 l i t e r s of water lday, man would i n g e s t 50 pg NTA per day. It can be ca l cu la t ed t h a t man would i n g e s t l e s s than 1.4 g NTA over a 75-year l i f e s p a n i f a l l d r inking water contained 25 pg NTA per l i t e r .

RISK IISSESSMENT The s c i e n t i f i c information a v a i l a b l e t o t h e Committee ind ica t ed no

apparent mutagenic o r t e r a t o g e n i c hazard from chronic NTA inges t ion . People i n Canada p r e s e n t l y i n g e s t l e s s than 0.001 mg N T A / ~ ~ bodyweightlday.

I n a s ses s ing t h e carcinogenic r i s k t o man from t h e present l e v e l s of NTA i n t h e environment we a r e faced wi th t h e usual u n c e r t a i n t i e s of e x t r a p o l a t i n g from extremely h igh experimental dose l e v e l s t o very low environmental l e v e l s '

of i n t e r e s t . The s t i l l g r e a t e r unce r t a in ty of e x t r a p o l a t i n g without any f i rm understanding of t h e underlying carcinogenic mechanism obviously makes any r i s k c a l c u l a t i o n very tenuous. Nevertheless , an at tempt has been made t o o b t a i n an e s t ima te of maximum r i s k us ing t h e d a t a a v a i l a b l e t o t h e Committee.

A l l r i s k c a l c u l a t i o n s have been based on t h e so-ca l led l i n e a r s t r a i g h t l i n e a r i t h m e t i c method which was proposed by t h e Subcommittee on Est imation of Risks of I r r e v e r s i b l e , Delayed Tox ic i ty t o t h e Department of Heal th, Education and Welfare Committee t o Coordinate Toxicology and Related Programs (Hoel, -- e t a l . ) u s ing a 95% confidence l e v e l f o r e x t r a p o l a t i o n . Ca lcu la t ions have been performed f o r each ind iv idua l dose l e v e l f o r a l l u r ina ry t r a c t tumors i n each s tudy. A zero background r a t e has been assumed.

This procedure produces an e s t ima te of t h e maximum r i s k a s soc i a t ed wi th any s p e c i f i e d dose, i f t h e t r u e dose response is concave upwards over t h e dose range of i n t e r e s t .

Table 5 con ta ins t h e r e s u l t s of t h i s e x t r a p o l a t i o n process f o r t h r e e environmental dose l e v e l s . I n a s se s s ing t h e s e r e s u l t s i t is important t o bear i n mind t h a t : ( a ) t h e r i s k f i g u r e r epo r t ed i n t h e t a b l e i s an e s t ima te of t h e maximum r i s k , t hus t h e a c t u a l r i s k could l i e w e l l below t h i s va lue ; and (b) t h a t , most impor tan t ly , t h e s e c a l c u l a t i o n s assume t h a t t h e t r u e response curve is concave upwards throughout t h e dose range under considera- t i o n , so t h a t a s t r a i g h t l i n e would produce an upper bound on t h i s curve - see f i g u r e below.

0 dose

DOSE RESPONSE COMPARISONS WITH SPECIFIC CARCINOGENS

(a) Carc inogenic i ty t o t h e Kidney of Af l a tox ins

Af la toxin B1 has been fed t o r a t s a t a l e v e l of 1 .0, 0.5, and .25 ppm f o r o r f i v e months and t h e animals were observed dur ing t h e i r l i f e s p a n f o r tumor product ion. Besides t h e high inc idence of l i v e r tumors 16 o u t of 45 animals a l s o showed r e n a l tumors a t a l l t h r e e dose l e v e l s (S. M. Eps te in , -- e t a l . ) . These observa t ions confirm e a r l i e r s t u d i e s by Bu t l e r and Barnes, who showed r e n a l adenocarcinomas o r carcinoma of t h e r e n a l p e l v i s i n 9% of t h e male r a t s fed a f l a t o x i n B a t t h e l e v e l of 0.5 - 0.1 ppm f o r l i f e . 1

Merkow i n a more r ecen t paper descr ibed r e n a l neop la s i a i n Wistar r a t s , fed 1 - 3 ppm of a f l a t o x i n , i n a l a r g e percentage of t h e s e animals.

From these s t u d i e s , i t can be concluded t h a t a f l a t o x i n B can cause 1 r e n a l cancer i n r a t s fed a d i e t of between 0.25 - 3 ppm of t h e carcino-

gen. I n t h e s e s t u d i e s l i v e r cancers were a l s o observed and i t i s , of course , w e l l known t h a t a f l a t o x i n is a very poten t hepatocarcinogen.

TABLE 5

ESTIMATED MAXIMUM RISK OF URINARY TRACT TUMOR FORMATION IN RODENTS

AT

LOW DOSES OF NTA OR Na3NTA

-

Dose (pg/kg ppb in diet)

5 25 5 0

Na3NTA

Male Rats (SRI) * Male Rats (LB)

Female Rats (SRI)

Female Rats (LB)

Male Mice (LB)

Female Mice (LB)

Male Rats (N1EHS)t

NTA -

Male Rats (LB)

Female Rats (LB)

Male Mice (LB)

Female Mice. (LB)

*The most. representative estimate of maximum risk has been selected for each study.

+Based on a dose of 1,000 ppm. in.-the water rather than diet, using a normal approximation' (cf . Hoel, -- et a1 's . Subcommittee's Report o n Risk Estimation) to take observed background into account .

The information t h a t a f l a t o x i n fed t o r a t s a t a l e v e l a s low a s 0.25 ppm can produce some kidney cancers and may reach a cancer incidence of 50% on feeding 3 ppm of a f l a t o x i n sugges ts t h a t t h e potency of t h i s carcinogen may be about 10,000 times g r e a t e r than t h a t of NTA, bu t i t would be unwise t o make such q u a n t i t a t i v e comparisons regard ing potencies i n humans. However, i t may se rve t h e purpose of i l l u s t r a t i n g some dose response comparisons of s p e c i f i c carcinogens.

(b) Carc inogenic i ty of Lead Compounds

Lead has been s tud ied f o r ca rc inogen ic i ty and was found t o be p o s i t i v e i n mice and r a t s . E a r l i e r experiments were f r equen t ly negat ive but many of them were inadequate.

P o s i t i v e r e s u l t s were obtained wi th l ead a c e t a t e and subace ta t e i n r a t and mouse feeding s t u d i e s . A d i e t con ta in ing 0.1% b a s i c lead a c e t a t e fed t o Swiss mice produced benign and malignant r e n a l tumors (Van Esch & Kroes). The same compound fed t o r a t s a t t h e 0.1 o r 1% l e v e l i n t h e d i e t a l s o produced t h i s type of tumor. (Van Esch, -- e t a 1 .). Renal tumors were a l s o observed i n r a t s fed d i e t s conta in ing 1% lead a c e t a t e . (Boyland, e t a l . ) Benign and malignant kidney tumors were observed i n r a t s fed l ead a c e t a t e ( 3 mg/day f o r 2 months and 4 mg/day f o r 16 months). Besides t h e kidney tumors,. neoplasms of t h e t e s t e s , ad rena l s , t hy ro id , p i t u i t a r y , and p r o s t a t e were observed i n t h e s e Wistar r a t s . (Zawirska & Medras) . I n male Sprague-Dawley r a t s on a d i e t con ta in ing 1% lead subace ta t e , 2 gliomas were observed bes ides 1 3 kidney tumors i n 17 animals. (Oyosu, e t a l . ) . Subcutaneous i n j e c t i o n s of l ead phosphate, repea ted over a 16 -- month per iod , produced adenomas, papi l lomas, and cystadenomas of t h e r e n a l c o r t e x i n a l b i n o r a t s . The t o t a l dose of lead administered v a r i e d between 120 and 680 mg Pb i n t h e animals producing tumors (Zo l l inge r ) .

N o p o s i t i v e f ind ings have been repor ted on humans.exposed t o high l e v e l s of l e a d , only nega t ive d a t a were repor ted by Dingwall-Fordyce and Lane; who followed 425 persons wi th a h i s t o r y of exposure t o l ead i n a b a t t e r y f ac to ry . They showed no i n c r e a s e i n cancer inc idence but an i n c r e a s e i n cerebrovascular acc iden t s w a s observed. This i s o f fe red as a b a s i s f o r t h e suggest ion t h a t s p e c i e s d i f f e r e n c e s i n t h e case of l ead t o x i c i t y a r e d i s t i n c t l y apparent . There a r e s u f f i c i e n t pub l i ca t ions on t h e ill e f f e c t s of l e a d exposure of humans t h a t ca ses of cancer i n these people, even i f anecdota l , would have been repor ted i n t h e l i t e r a t u r e over t h e years . A r ecen t s tudy of m o r t a l i t y among more than 7,000 l ead workers a l s o shows no inc rease i n r e n a l tumors (Cooper & Gaffey) .

When t h e experimental d a t a on rodents a r e used f o r e x t r a p o l a t i o n t o human exposure, t h e equiva lent dose f o r r e n a l tumor development , i n man would be 550 mg l ead per day, which appears t o exceed t h e maximum t o l e r - a t e d dose of l ead f o r man (IARC Monographs).

Afghan, B. K . and P. D . Goulden. Determination of Trace Quan t i t i e s of N i t r i l o t r i a c e t i c Acid by D i f f e r e n t i a l Cathode - Ray Polarography, Env. Sci . Technol., (1971), - 5 , 601.

Afghan, B. K . , P. D. Goulden and J. F:Ryan. Automated Method f o r Determination of N i t r i l o t r i a c e t i c Acid i n Natural Water, Detergents and Sewage Samples, Anal. Chem., (1972), 44, (2 ) , 354-359.

Ambient NTA Levels Seem Harmless, Even Benef i c i a l t o Aquatic Systems, Quar ter ly Report of t h e Environmental Research Laboratory - Duluth,, 6201 Congdon Blvd., July-September (1976), pp. 4-5, U . S. Environmental . ' Pro tec t ion Agency, Of f i ce of Research and Development. -

Arthur, J . W . , A. E. Lemke, V . R . Mattson and B. J. Hal l igan , Toxic i ty of Sodium N i t r i l o t r i a c e t a t e (NTA) t o t h e Fathead Minnow and an Amphipod i n Sof t Water, Water Research, (1974), - 8 , ( 3 ) , 187-193.

Ashforth, G. K. and G. Calvin. Safe ty Evaluat ion of S u b s t l t u t e s f o r Phosphates i n Detergents , Water Research, (1973), - 7, (1-2) 309-320.

Assessment of t h e P o t e n t i a l of N i t r i l o t r i a c e t a t e (NTA) t o Compromise Human Health; Ass i s t an t Sec re t a ry ' s Ad Hoc Group on NTA, F ina l Report t o t h e Ass i s t an t Sec re t a ry f o r Health and S c i e n t i f i c A f f a i r s , Dept. of Heal th, Education and Welfare, Apr i l 19 , 1972.

Bagley, D. H . , J r . , E. J. Zapolski , M . Rubin and J. V . P r i n c i o t t o . The Metabolism and P l a c e n t a l Transfer of Oral and P a r e n t e r a l I ron Chela tes , C l in i ca Chimica Acta, (1971), - 35, (2 ) , 311-315.

Banat, K . , U . Fgrs tner and G . M u l l e r . Experimental Mobilizat ' ion of Metals from Aquatic Sediments by N i t r i l o t r i a c e t i c Acid, Chemical Geology, (1974), - 14, 199-207.

Baner j i , S. K. L i t e r a t u r e Review: Detergents . J.W.P.C.F., (1973), 45, (6) , 1059-1063 .. -

Barica, J . , M. P. S t a in ton and A . L. Hamilton: Mobi l iza t ion of Some Metals i n Water and Animal Tissue by NTA, EDTA and TPP, Water Research, (1973), 7 , (12) , 1791-1804. -

Bender, M. E . , W . R. Matson and R. A. Jordan. On t h e Signi f icance of Metal Complexing Agents i n Secondary Sewage E f f l u e n t s , Env. Sc i . Technol., (1970), - 4, 520.

Bies inger , K. E . , R . W . Andrew and J. W. Arthur , Chronic Toxic i ty of NTA ( N i t r i l o t r i a c e t a t e ) and Metal-NTA Complexes t o Daphnia magna, J. Fish. Res. Board Can., (1974), - 31, (4) , 486-490.

Bies inger , K. E . , and G. M. Chr i s t iansen . E f fec t of Various Metals on t h e Surv iva l , Growth, Reproduction and Metabolism of Daphnia magna, J. F ish . Res. Board Can., (1972), - 29, 1691.

Bioassay of NTA and Na NTA. H 0 f o r P o s s i b l e Carc inogenic i ty , December 1976. Z Unpublished r e p o r t of ?he Nat ona l Cancer I n s t i t u t e .

Bjoerndal , H . , H . 0. Bouveng, P. Solyom and J . Werner. NTA i n Sewage Treatment. P a r t 3. Biochemical S t a b i l i t y of Some Metal Chela tes , Vat ten, (1972), 28, ( I ) , 5-6.

Bora, K. C. E f f e c t s of N i t r i l o t r i a c e t i c Acid (NTA) on Chromosome Rep l i ca t ion and S t r u c t u r e i n Human C e l l s , Mutat. Res., (1975), - 31, 325.

Bourquin, A. W . , and V . ,A. Przybyszewski. D i s t r i b u t i o n of Bac te r i a w i th N i t r i l o t r i a c e t a t e Degrading P o t e n t i a l i n an Es tua r ine Environment, unpublished r e p o r t , U.S. EPA Environmental Research Laboratory, Gulf Breeze, F lo r ida .

Boyland, E . , C. E. Dukes, P. L. Grover and B. C . V . Mitchley. The Induct ion of Renal Tumours by Feeding Lead Aceta te t o Rats , B r i t . J. Cancer, (1962), 16, 283.

Budny, J. A. Metabolism and Blood P res su re E f f e c t s of Disodium N i t r i l o t r i a c e t a t e (Na2NTA) i n Dogs, Toxicol . Appl. Pharmacol., (1972), - 22, ( 4 ) , 655-660.

Budny, J. A., and J. D . Arnold. N i t r i l o t r i a c e t a t e (NTA): Human Metabolism and i t s Importance i n t h e T o t a l Sa fe ty Evaluat ion Program, Toxicol . Appl. Pharmacol., (1973), 25, ( I ) , 48-53.

Budny, J. A., R. J . Niewenhuis, E. V. Buehler and E. I. Goldenthal. Subacute Ora l Tox ic i ty of Trisodium N i t r i l o t r i a c e t a t e (Na3NTA) i n Dogs, Toxicol . Appl. Pharmacol . , ( l973) , - 26, ( I ) , 148-153.

~ i r g i , H. The E f f e c t s of NTA on t h e Growth o f Phytoplankton wi th P a r t i c u l a r Considerat ion Given t o I r o n a s a Microelement, Schweiz. , Z . Hydrol. , (1974), 3 6 , '(I),, 1-70. -

Bunch, R. L., and M. B. E t t i nge r . Biodegradabi l i ty of P o t e n t i a l Organic S u b s t i t u t e s f o r Phosphates, Proc. 22nd. Ind. Wastes Conf., 1967, Purdue Un ive r s i t y , p. 393.

Busch, E. A. B io log ica l Assessment of t h e Tox ic i ty of Organic Complexing Agents by Means of Protozoa and Lower Metazoa, Z . Wasser, Abwasser Forsch., (1974), L, ( 3 ) , 65-91.

B u t l e r , W. H . , and J. M. Barnes. Fd. Cosmet. Toxicol . , (1968), - . 6 , 135-141.

Carey, J. H. Photochemical Degradation of NTA, Nat iona l Research Council of Canada, Post-Doctoral Fellow, Water Chem. Sec t ion , Canada Centre f o r In land Waters, Bur l ing ton , Ontar io - personal communication t o D. H . Matheson.

Carey, J. H. and C. H. Langford. Photodecomposition of Fe (111) Aminopoly- carboxyla tes , Can. J. Chem., (1973), 51, 3665. -

Chau, Y . K . , R. ~ g c h t e r and K. Lum-Shue-Chan. ~ e t e r m i n a t i o n of t h e ~ p ~ a r e n t -

Complexing Capac i ty of Lake Waters, J . F i sh . Res. Board Can. , (1974) , 31, 1515.

Chau, Y. K . , and M. T. Shiomi. Complexing P r o p e r t i e s of N i t r i l o t r i a c e t i c Acid i n t h e Lake Environment, Water, A i r and Sewage P o l l n . , (1972) , L, 149.

Ch i l d s , C. W . Chemical Equi l ib r ium Models f o r Lake Waters Which Contain N i t r i l o t r i a c e t a t e and f o r "Normal" Lake Water, Proc. 1 4 t h . Conf. Great Lakes Res., (1971), 198.

C h r i s t i e , A. E. Trisodium N i t r i l o t r i a c e t a t e and Algae, Water and Sewage Works, (1970) , - 117, 58.

Chu, I . , D. C. V i l l eneuve and G . C . Becking, (1976) - unpubl i shed d a t a .

C i l l e y , W. A . , and D. A. Nicholson. An Equi l ib r ium Model f o r t h e NTA Complexation o f Metal I o n s i n Na tu r a l Waters, Envt l . L e t t e r s , (1971), 2, ( 3 ) , 121.

Claesson, A. Anaerobic B a c t e r i a l , Degradat ion of N i t r i l p t r i a c e t a t e , NTA, Va t t en , (1971), 27, ( 4 ) , 410-411.

Cleasby, J. L . , D . W. Hubly, T. A. Ladd and E. A. Schon. T r i c k l i n g F i l t r a t i o n of a Waste Conta in ing NTA, J.W.P.C.F., (1974) , - 46 , ( 8 ) , 1873-1887.

"Commentary" - Report on Carc inogenes i s ~ i o a s s a y s of N i t r i l o t r i a c e t i c Acid and N i t r i l o t r i a c e t i c Acid, Trisodium S a l t , Monohydrate; Carc inogenes i s Program, D iv i s i on of Cancer Cause and P reven t i on , Na t i ona l Cancer I n s t i t u t e , June 1, 1976.

Cooper, W. C . , and W. R. Gaffey. M o r t a l i t y of Lead Workers, J . of Occup. Medicine, (1975) , - 17 , 100-107.

David, J . A . , and J. Jacknow. Heavy Meta l s i n Waste Water i n Three Urban Areas, J.W.P.C.F., (1975) , - 47, 2292.

Dingwall-Fordyce, I . , and R. E. Lane. A Follow-up Study of Lead Workers, B r i t . J . I n d u s t r . Med., (1963), 20, 213.

Doig, Marion T., 111, and Dean F. Mart in . A Note Concerning t h e Environmental A c c e p t a b i l i t y o f N i t r i l o t r i a c e t i c Acid (NTA): t h e E f f e c t of NTA on t h e Growth of Gymodinium breve, Env t l . L e t t e r s , (1974), 5, ( I ) , 31-36.

Dunlap, W. J . , R. L. Cosby, J. P. McNabb, B. E. Bledsoe and M. R. Sca lp . Probab le Impact of NTA on Ground Water, Ground Water, (1972), 10 , (I), 107-117. - Eden, G. E . , G . F. Cul ly and R. C . Rootham. E f f e c t of Temperature on t h e Removal of NTA During Sewage Treatment , Water Research , (1974) , - 6 , 877.

E i s l e r , R . , G. R. Gardner, R. J. Hennekey, G. Laroche, D . F. Walsh and P. P. Yevich. Acute T o x i c i t y of Sodium N i t r i l o t r i a c e t i c Acid (NTA) and NTA-Containing De t e rgen t s t o Marine Organisms, Water Research, (1972), 6 , ( 9 ) , 1009-1027. -

El-Masry, A , , A. Ghazal, M. E. El-Sayed and H. M. Farag. Pharmacological S tudies of t h e I n t e r a c t i o n s Between Some Ant id i abe t i c s and t h e Calcium Chelat ing Agent N i t r i l o t r i a c e t i c Acid i n t h e Rabbit , Egypt. J. Pharm. S c i . , (1972), 13, ( I ) , 23-37.

Enfors, S. - 0. and N. Molin. Anaerobic Degradation of N i t r i l o t r i a c e t a t e (NTA) by Bacter ia , Vatten, (1971), 27, ( 2 ) , 162-163.

Enfors, S. - 0. and N. Molin. Biodegradation of N i t r i l o t r i a c e t a t e (NTA) by Bacter ia - I. I s o l a t i o n of Bac te r i a Able t o Grow Anaerobical ly wi th NTA a s a Sole Carbon Source, Water Research, (1973), - 7, ( 6 ) , 881-888.

Enfors, S. - 0. and N. Molin. Biodegradation of N i t r i l o t r i a c e t a t e (NTA) by Bacter ia - 11. C u l t i v a t i o n of an NTA-degrading Bacterium i n Anaerobic Medium, Water Research, (1973), 1, ( 6 ) , 889-893.

Environmental P ro tec t ion Agency (U.S.), Great Lakes Water Qual i ty : S t a t u s of Per t ine l i t EPA Research, EPA 66019-74-002.

Epstein, S. M . , B. Bartus and E. Farber. Cancer Res., (1969), - 29, 1045-1050.

Epstein, S. S. Toxicologica l and Environmental Impl ica t ions on t h e Use of N i t r i l o t r i a c e t i c Acid a s a Detergent Builder - I, I n t . J. Environ. S tud . , (1972), 2, ( 4 ) , 291-300.

Epstein, S. S., Toxicologica l and Environmental Impl ica t ions on t h e Use of N i t r i l o t r i a c e t i c Acid a s a Detergent Builder - 11, I n t . J. Environ. Stud. , (1972), - 3, ( I ) , 1'3-21.

Erickson, S. J . , T. E. Maloney and J. H. Gent i le . E f f e c t s of N i t r i l o t r i a c e t i c Acid on t h e Growth and Metabolism of Es tuar ine Phytoplankton, J.W.P.C.F., (1970), 42, R329.

F i res tone , M. K. and J. M. T iedje . Biodegradation of M e t a l - N i t r i l o t r i a c e t a t e Complexes by a Pseudomonas Species: Mechanism of React ion, Appl. Microbiol . , (1975), - 29, (6 ) , 758-764.

Flannagan, J. F. Tox ic i ty Evaluat ion of Trisodium N i t r i l o t r i a c e t a t e t o Se lec ted Aquatic I n v e r t e b r a t e s and Amphibians, F i s h e r i e s Research Board of Canada Technological Report No. 258, (1971), 1-15.

Flannagan, J. F. In f luence of Trisodium N i t r i l o t r i a c e t a t e on t h e Mor ta l i t y , Growth and Fecundity of t h e Freshwater S n a i l (HeZisoma t r i v o l i s l Through Four Generations, J. Fish. Res. Board Canada, (1974), - 31, ( 2 ) , 155-161.

Flynn, F. G. and P. H. Jones. The Environmental and Hygenic,Safety of N i t r i l o t r i a c e t a t e (NTA), Proc. of t h e S ix th Canadian Symp. on Water Pol ln . Research, Toronto, February 1971, 271 pp.; 13-37.

Focht, D. D. and H. A. Joseph. B a c t e r i a l Degradation of N i t r i l o t r i a c e t i c Acid (NTA), Can. J. Microbial., (1971), 17 , (12) , 1553-1556. - -

Focht, D. D. and H. A. Joseph. Nitrilotriacetic Acid Abatement of Heavy Metal Toxicity Towards Pseudomonas, Abstr. Ann. Meet. Am. Soc. Microbial., (1972), 72, 6.

Garry, V. P. and E. M. Oyeki. Detergent Effects on the Development of Antibody Producing Cells -- in vitro. Model for the Study of Chemical Effects on Cell Differentiation in a Developing System, Oncology, (1973), 27, (2), 160-167.

Gilbert, C., J. Gillman, P. Loustalot and W. Lutz. The Modifying Influence of Diet and the Physical Environment on Spontaneous Turnover Frequency in Rats, The British Journal of Cancer, (1958), - XII,565-593.

Golterman, H. L. Natural and Accelerated Phosphate Mobility, Geol. Mijnbouw, (1974), 53, (4), 205-206.

Goyer, R. A. and K. R. Mahaffey. Metabolic and Renal Effects'of Oral Trisodium Nitrilotriacetate (NTA) Ingestion in Control and Lead-Fed Rats, Fed. Proc., (1972), - 31, (2), 638..

Greenblatt, M. and W. Lijinsky. Carcinogenesis and Chronic Toxicity of Nitrilo- triacetic Acid in Swiss Mice, J. of the National Cancer Institute, (1972j, 2, (4), 1123-1126.

Gregor, C. D. Solubilization of Lead in Lake and Reservoir Sediments by NTA, Environ. Sci. Technol., (1972), - 6, (3), 278-279.

Gudernatsch, H. Biologischer Abbau von ~itrilotriessigs~ure, Wasser abwasser, (1974), - 115, (9), 418-421.

Gutzke, E. R., J. R. Reed and G. C. Grant. The Influence of Nitrilotriacetic Acid on Heavy Metal Ion Uptake in Blue Gills (Lepomis macrochirus), Va. J. Sci., (1973), 24, (3), 145.

Hamilton, R. D. NTA: Environmental Acceptability, Nutrients and Eutrophication Symposium, (1971), 1, 217.

Hoel, -- et al. Subcommittee on Estimation of Risks of Irreversible, Delayed Toxicity Report to the U.S. Department of Health, Education and Welfare Committee to Coordinate Toxicology and Related Programs.

Hoover, T. B. Polarographic Determination of NTA: Environmental Protection Technology Series, Monitoring Agency Rept. No. W 72-13665, EPA-R2-73-254; PB 222 940/9; Proj. EPA-16020-EWE; 53 pp., June 1973.

Huber, L. Studies on the Biodegradability of Nitrilotriacetic Acid (NTA) , Tenside Detergents, (1974), - 11, (I), 17-26. The Bayer Biological Research ' '

Institute, Munich.

IARC Monographs on the Evaluation of Carcinogenic Risk of Chemicals-to Man, IARC, Lyon, France, (1972), 1, 40-50.

Industrial BIO-TEST Laboratories, Inc., 1810 Frontage Road, Northbrook, Ill. Report to Monsanto Company, Teratogenic Study with Methylmercuric Hydroxide, Methylmercuric Chloride or Cadmium Chloride in Combination with Synthetic or Natural Chelants, June 24, 1971. IBT No. B162.

Industrial BIO-TEST Laboratories, Inc., 1810 Frontage Road, Northbrook, Ill. Addendum Report to Government/Industry Committee, Internal and Skeletal Examination of Fetuses Obtained from the Teratogenic Study with Methylmercuric Chloride or Cadmium Chloride in Combination with Synthetic or Natural Chelants in Albino Rats, August 10, 1971. IBT No. B327.

Investigation of the Mutagenic Effects in Mice of Nitrilotriacetic Acid (NTA) by the Dominant Lethal Gene Test, Report from Stanford Research Institute, June, 1971.

IJC Task Force Presentation on NTA Health Implications by the Procter & Gamble Company and Monsanto Industrial Chemicals Company,' October 14, 1976, Cincinnati, Ohio - Unpublished. Jacobson, K. H. Effect of Nitrilotriacetic Acid Chelation on the Subacute Oral Toxicity in Rats of Cadmium and Methyl Mercury, Toxicol. Appl. Pharmacol., (1972), - 22; (2), 296.

Jaques, L. B. and T. K. Sue. Mobilization of Chondroitin Sulphate by Nitrilotriacetic Acid, Toxicol. Appl. Pharmacol., (1975), - 34, (3), 521-524.

Jarrett, C. L. and L. B. Jaques. Effect of I.V. and Oral Administration of Heparinoids G31150, G3115-A and of Nitrilotriacetic Acid on Blood Coagulation, Thromb. Diath. Haemorrh. , (1971) , - 25, (1) , 187-200.

Jenkins, D., W. J. Kaufman, P. H. McGauhey, A. J. Horne and J. Gasser. Environmental Impact of Detergent Builders in California Waters, Water Research, (1973), 1, 265.

Jolley, R. L. Chlorination Effects on Organic Constituents in Effluents from Domestic Sanitary Sewage Treatment Plants, Univ. of Tennessee Ph.D. Thesis, (1973).

Jorgenson, E. A., G. W. Newell, L. G. Scharpf, P. Gribling, M. 0. O'Brien and D. Chu. Study of the Mutagenic Potential of the Monocalcium. Salt of Nitrilo- triacetic Acid (Na Ca NTA) bythe Dominant Lethal Test in Mice, Stanford Research Institute, Menlo Park, California; Monsanto Industrial Chemicals Co., St. Louis, Missouri.

Jorgenson, E. A., G. W. Newell, L. G. Scharpf, P. Gribling, M. 0. 0'Brien and D. Chu. Study of the Mutagenic Potential of Nitrilotriacetic Acid (Na Ca NTA) in Mice.by the Translocation Test, Stanford Research Institute, Menlo Park, California; Monsanto Industrial Chemicals Co., St. Louis, Mo.

Kaiser, K. L. E. Determination and Differentiation of Ethylenediaminetetra- acetic Acid (EDTA) and Nitrilotriacetic Acid (NTA) in Freshwater, water Research, (1973), - 7, (lo), 1465-1473.

Kamat, M. H., P. J. Mennert and J. J. .Seebode. Effect of Oral Polyphosphate and Nitrilotriacetic Acid on Experimental Rat Urolithiasis, Surg. Forum., (1971), - 22, 12-13.

Kania, H. J., and H. J. Beyers. NTA and Mercury in Artificial Stream Systems, EPA Ecol. Res. Series EPA-66013-73-025, 25 pp., Feb. 1974.

Klein, S. A. NTA Removal in Septic Tank and Oxidation Pond Systems, J.W.P.C.F., (1974), - 46, (I), 78-88.

Kolonel, L. N. Association of Cadmium with Renal Cancer, Cancer,-(1976), 37, (4), 1782-1787.

Kramers, P. G. N. Mutagenicity Studies with Nitrilotriacetic Acid (NTA) and Citrex S-5 in Drosphila, Mutation Research, (1976), - 40, 277-280.

Laham, S., J. M. F. Rajabalee and M. Potvin. Thin Layer Chromatography.of . . .

Chelates of Nitrilotriacetic Acid, Assoc. Can. Pr. Av. Sci., (1972), - 3.9, 1-43. . .

Langford, C. H., M. Wingham and V. S. Sastri. Ligand Photoxidation in Copper (11) Complexes of Nitrilotriacetic Acid; Implications for Natural Waters, Env. Sci. Technol., (1973), - 7, 820.

Laroche, G., G. R. Gardner, R. Eisher, E. H. Jackim, P. P. Yevish and G. E. Zaroogian. Analysis of Toxic Responses in Marine Poikilotherms, Bioassay Tech. Environ. Chem., (1973), 199-216.

Larson, H. A., J. C. Weston and S. M. Howell. Quantitative Gas Chromatographic Determination of Nitrilotriacetic Acid in the Presence of Other Carboxylic Acids, J. Chromatograph. , (1975) , 111, (I), 43-50.

Lautenbacher, H. W., and H. W. Baker. Determination of NTA (nitrilotriacetic acid) in Aquatic Toxicity Studies. Compleximetric Method Using Atomic Absorption Spectroscopy, Bull. Environ. Contam. Toxicol., (1974), - 11, (I), 57-63.

Lawson, D. W. National NTA Monitoring Program. Progress Report No. 2, (1972);

Lebish, Capt. I. J., Lt. Col. D. G. Simons, H. Yagoda, P. Janssen and W. Haymaker. Observations on Mice Exposed to Cosmic Radiation in the Strato- sphere, a Longevity and Pathological Study of 85 Mice, Military Medicine, (1959) , - 24, (12), 835-847.

Lekokmakher, S. S. Behaviour of Zinc, Calcium and Manganese Chelates in Animals, Tr. Instr. Ekol. Rast. Zhivotn., Ural. Nauchn. Tsentr., Akad. Nauk. SSSR, (1974), - 89, 67-71.

Lijinsky, W., M. Greenblatt and-C. Kommineni. Brief Conmrunication: Feeding Studies of Nitrilotriacetic Acid and Derivatives in Rats, Journal of the National Cancer Institute, (1973), - 50, (4), 1061-1063.

Litchfield, C. D. and R. P. Huber. Effect of Selected Pollutants on the Specific Growth Rate and Extracellular Enzyme Synthesis of Aeromonas proteolytica, Bull. Ecol. Res. Comm., NPR, (1973), 17, 464-466.

Little, Arthur D., Inc. Report - See Thayer & Kensler.

Liu, D., P. T. Wong and B. J. Dutka. Studies of a Rapid NTA Utilizing Bacterial Mutant, J.W.P.C.F., (1973), 45, (8), 1728-1735.

Longman, G. F., M. J. Stiff and D. K. Gardiner. The Determination of Nitrilotriacetic Acid in Sewage and Sewage Effluent, Water Research, (1971), 5, (121, 1171-1175. -

~uning, K. G., J. Lindsten, K. Lindstrom, W. Sheridan, M. Multen and T. Linne. Personal Communication to Monsanto Industrial Chemicals Co. from S. S. Epstein.'~ Staff Report to Committee on Public Works, U.S. Senate, Dec. 1970, in .Int.'J. Environ. Studies, (1972), 2, 291.

Macek, K. J., and R. N. Sturm. Survival and Gill Condition of ~luegill (Lepomis macrochirces) and Fa thead Minnows (Pimepha Zes prome Zas) , Exposed to Sodium Nitrilotriacetate (NTA) for 28 Days, J. Fish. Res. Board Can., (1973), 30, (2), 323-325. -

Maguire, J. Kinetics of the Cu (11)-nitrilotriacetic Acid Reaction, Can. J. Chem., (1974), 52, 4106.

Mahaffey, K. L., and R. A. Goyer. Trisodium Nitrilotriacetate in Drinking Water: Metabolic and Renal Effects in Rats, Arch. Environ. Health, (1972), 25, (4), 271-275.

Malcolm, A. R., B. H. Pringle and H. W. Fisher. Chemical Toxicity Studies with Cultured Mammalian Cells, Bioassay Tech. Environ. Chem., (1973), 217-230.

Manahan, S. E., and M. J. Smith. The Importance of Chelating Agents in Natural Waters and Wastewaters, Water and Sew. Wks., (1973), 120, 102.

-Mancini, P. :Use of Nitrilotriacetic Acid (NTA) in Synthetic Detergent Formulations, Ross. Chim., (1975), 27, (4), 163-167.

Martin, A. E. Medical Considerations in the Abstraction of Potablewaters from Polluted Sources, Water Treatment and ~xamination, (1972), 21, 202.

Martin, D. F. Biostimulatory Effect of Nitrilotriacetic Acid on Growth and Photosynthetic Rate of the Red Tide Dinoflagellate GonyauZm tamarensis, Comments, Environ. Letters.?(l974), 1, 175-177.

Matheson, D. H. A Report on ~itrilotriacetic Acid in the Canadian Environment, unpublished report prepared for the Canada Centre for Inland Waters, Burlington, Ontario.

Merkow, L. P., et al. Cancer Res., (1973), 33, 1608-1614.

Michael, W. R., and J. M. Wakim. Metabolism of Nitrilotriacetic Acid (NTA), Toxicol. Appl. Pharmacol. (1971), 18, (2), 407-416.

Michael, W. R., and J. M. Wakim. Calcium and Zinc Metabolism in the Presence of Trisodium Nitrilotriacetate, Toxicol. Appl. Pharmacol., (1972), 22, (2), 297.

Michael, W. R., and J. M. Wakim. Effect of Trisodium Nitrilotriacetate (Na NTA) on the Metabolism of Selected Metal Ions, Toxicol. Appl. Pharmacol., (1913)~ 2, (4), 519-529.

Mottola, H. A. Nitrilotriacetic Acid as a Chelating Agent: Applications, Toxicology and Bio-Environmental Impact, Toxicological and Environmental Chemistry Reviews, (1974), 2, (2), 99-161.

Mottola, H. A., and G. L. Heath. Detection and Variable Time Kinetic Determination of Micro and Submicrogram Amounts of Nitrilotriacetic Acid, Anal. Chem., (1972), - 44, (14), 2322-2325.

Muggenburg, B. A., J. L. Mauderly, F. F. Hahn, S. A. Silbaugh and S. A. Felicetti. Effects of the Ingestion of Various Commercial ~eter~ent' Products by Beagle Dogs and Pigs, Toxicol. Appl. Pharmacol., (1974), 30, (I), 134-148.

Mugwira, L. M., B. D. Knezek and B. G. Ellis. Effect of Chelates on Metal Activation from River and Lake Sediments, Ann. Mtg. Am. Soc. of Agron., (Abstract), Aug. 15-20, 1971.

Munawar, M., and W. A. Glooschenko. The Effects of NTA Upop Primary Production by -- in situ Phytoplankton in Lakes Erie and Ontario, Unpublished Manuscript, Canada Centre for Inland Waters, Burlington, Ontario.

Murillo, A., M. S. Campos and G. Varela. Factors Affecting the Digestibility, Absorption and Retention of Calcium. Effect of Oxalate, Ethylene-diaminete- tracetic Acid Disodium Salt, Nitrilotriacetic Acid, Lysine, and Protein Quality, Rev. Espan. Fisiol., (1972), 28, (2), 115-23.

Murphy, K. L., R. Zaloun and D. Fulford. Effects of Chlorination Practice on Soluble Organics, McMaster Univ., Hamilton, Ontario, Dept. of Chem. Eng., 73-1211-4, (1973).

Nilsson, R. Removal of Heavy Metals by Chemical Treatment of Municipal Wastewater, Water Research, (1971), 2, 51.

Nixon, G. A. Toxicity Evaluation of Trisodium Nitrilotriacetate, Toxicol. Appl Pharmacol., (1971), 18, (2), 398-406.

Nixon, G. A., E. V. Buceler and R. J. Niewenhuis. Two-Year Rat Feeding Study with Trisodium Nitrilotriacetate and its Calcium Chelate, Toxicol. Appl. Pharmacol., (1972), 21, (2), 244-252.

Nolen, G. A., R. L. Bohne, and E. V. Buehler. Effects of Trisodium Nitrilotriacetate, Trisodium Citrate and a Trisodium Nitrilotriacetate-Ferric Chloride Mixture on Cadmium and Methyl Mercury Toxicity and Teratogenesis in Rats, Toxicol. Appl. Pharmacol., (1972), 23, (2), 238-250.

Nolen, G. A., E. V. Buehler, R. G. Geil and E. I. Goldenthal. The Effects of Trisodium Nitrilotriacetate on Cadmium and Methyl Mercury Toxicity and Terato- genicity in Rats, Teratology, (1972), 5, (2), 264.

Nolen, G. A., E. V. Buehler, R. G. Geil and E. I. Goldenthal. Effects of Trisodium Nitrilotriacetate on Cadmium and Methyl Mercury Toxicity and Teratogenicity in Rats, Toxicol. Appl. Pharmacol., (1972), 23, (2), 222-237.

Nolen, G. A., E. V. Buehler, R. G. Geil and E. I. Goldenthal. Effects of Trisodium Nitrilotriacetate, Trisodium Citrate and a Trisodium Nitrilotri- acetate-Ferric Chloride Mixture on Cadmium and Methyl Mercury Toxicity and Teratogenesis in Rats, Toxicol. Appl. Pharmacol., (1972), 23, (2), 238.

Nolen, G. A., L. W. Klusman, D. L. Back and E. V. Buehler. Reproduction and Teratology Studies of Trisodium Nitrilotriacetate in Rats and Rabbits, Fd. Cosmet. Toxicol., (1971), 9, (4), 509-518.

Novosielski, O., bD. Knezek and B. G. Ellis. Evaluation of Toxicity of Metals, NTA and EDTA by AspergiZZus,niger Bioassay, Amer. Chem. Soc., Div. Water, Air Water Ch., (1971), 11, (2), 32-38- General Paper.

Oyasu, R., H. A. Battifora, R. A. Clasen, J. H. McDonald and G. M. Hass. Inductionof Cerebral Gliomas in Rats with Dietary Lead Subacetate and 2-Acetylaminofluorene, Cancer kesearch, (1970), 3, 1248.

Palmer, M. D., et al. Hamilton Harbour Study, Ontario Ministry of the Environment, Toronto, (1974).

Parks, S. Mj., and P. E. Stukus. The-Microbial Metabolism of Nitrilotriacetic Acid, J. Sci. Lab. Denison Univ., (1973), 54, (3-6), 79-86.

Pickaver, A. H. The Production of N-nitrosoiminodiacetate from Nitrilotriacetate and Nitrate by Microorganisms Growing in Mixed Culture, Soil Biol. Biochem., (1976), 8, (I), 13-17.

Prakash, A. NTA (Nitrilotriacetic Acid) - An Ecological Appraisal, National Research Council of Canada, Ottawa, NRCC No. 15023, (1976), 46 pp; Associate Committee on Scientific Criteria for Environmental Quality.

Prejean, J. D., J. C. Peckham, A. E. Casey, D. P. Griswold, E. K. Weisburger and J. H. Weisburger. Spontaneous Tumors in Sprague-Dawley Rats and Swiss Mice, Cancer Research, (1973), 33, 2768-2773.

Procter & Gamble Company. Nitrilotriacetic Acid (NTA) Levels in Canadian Waters. Drinking Water Survey, Progress Report No. 1 (Nov. 1971 to July 1973).

Procter 6 Gamble Company. New York State (Long Island Study) 1972. Mimeograph Dated April 10, 1974.

Rajabalee, F. J. M., S. Laham and M. Potvin. Chemical Studies on Environmental Pollutants; thechelates of Nitrilotriacetic Acid with Alkaline Earth Metals, Chemosphere, (1973), 2, 19-22.

Renn, C. E. Biodegradation of NTA Detergents in a Wastewater Treatment System, J.W.P.C.F., (1974), - 46, (lo), 2363-2371.

36

Rudd, J. W. M. and R. D. Hamilton. Biodegradation of Trisodium N i t r i l o t r i a c e t a t e i n a Model Aerated Sewage Lagoon, J . Fish. Res. Board of Canada, (1972), 3, (8) , 1203-1208.

Rudd, J. W. M. , B. E. Townsend and R. D. Hamilton. Discharge of N i t r i l o - t r i a c e t a t e (NTA) from Two Sewage Treatment F a c i l i t i e s Located i n a Mid-Continental Climate, J . Fish. Res. Board of Canada, (1973), 30, 1026. -

Sanchez, I . , and G . F. Lee. Sorpt ion of Copper on Lake Monona Sediments: E f f e c t s of NTA on Copper Release from Sediments, Water Research, (1973), 1, (4 ) , 587-593.

Sass , B . , L. S. Rabstein, R. Madison, R. M. N i m s , R. L . P e t e r s and G . J. Ke l lo f f . Incidence of Spontaneous Neoplasms i n F344 Rats Throughout t h e Natural Life-Span, Journa l of t he National Cancer I n s t i t u t e , (1975), 54, (6 ) , 1449-1455.

Scharpf, L. G . , Jr . , I. D. H i l l , P. L. Wright and M. L. Keplinger. Teratology Studies on Methylmercury Hydroxide and Sodium N i t r i l o t r i a c e t a t e i n Rats, Nature (London), (1973), 241, 5390, 461-463.

Scharpf, L. G . , Jr., I. D. H i l l , P. L. Wright, J. B. Plank, M. L. Keplinger and J. C. Colandra. E f f e c t s of Sodium N i t r i l o t r i a c e t a t e orl Toxic i ty , Terato- g e n i c i t y and Tissue D i s t r i b u t i o n of Cadmium, Nature, (London), (1972), 239, 5369, 231-234.

Scharpf, L. G . , Jr., F. J . Ramos and I. D. H i l l . In f luence of N i t r i l o t r i a c e t a t e (NTA) on the Toxic i ty , Excre t ion and D i s t r i b u t i o n of Cadmium i n Female R a t s , Toxicol. Appl. Pharmacol. , (1972), 22, ( 2 ) , 186-192.

Shannon, E. E . , P. J. A. Fowlie and R . J. Rush. A Study of N i t r i l o t r i a c e t i c Acid (NTA) Degradation i n t h e Receiving Stream, Environment Canada, EPS 4-WP-74-7, (1974).

Shannon, E . E. , and L. J. Kamp. Detergent S u b s t i t u t i o n S tud ie s a t Canadian Forces S t a t i o n , Gloucester , Environment Canada, EPS 4-WP-73-3, (1973).

Sher, S. P. Tumors i n Control Mice: L i t e r a t u r e Tabulat ion, Toxicol. Appl. Pharmacol. , (1973), - 30, 337-359.

Shepard, T. H . N i t r i l o t r i a c e t a t e i n ~ e t e r ~ e n t s and Human Health, Teratdlogy, (1972), 6, (2) , 127-128.

Shumate, K. S., J. E. Thompson, J . D. Brookhart and C. L. .Dean. NTA Removal by Activated Sludge--Field Study, J.W.P.C.F., (1970), 42, (4 ) , 631-640.

Sprague, J. B. Promising Ant i -Pol lu tant ; Chelat ing Agent NTA P r o t e c t s F ish from Copper and Zinc, (1968), Nature, - 220, 1345. ( F i s h e r i e s Research, .Board. of Canada Study No. 1291).

S t ine , G . J. Ef fec t s of N i t r i l o t r i a c e t i c Acid on Development of ~ e u r o s ~ o r a crassa, Abstr . Ann. Meet. Am. Soc. Microbiol. , (1973), 73, 31.

S t i n e , G. J . , and P. Adams. E f f e c t s of N i t r i l o t r i a c e t i c Acid on Development of Neurospora crassa.

Stine, G..J., and A. A. Hardfgree. Effects of Nitrilotriacetic Acid (NTA) by the Domlnant Lethal Gene Test, Report from Stanford Research Institute, June, 1971.

Stine, G. J., A. A. Hardigree. Effect of Nitrilotriacetic Acid on Growth and Matting in Strains of Escherichia coZi K-12, Can. J. Microbiol, (1972), 18, (7), 1159-1162.

. . . .

Stolzberg, R. J., and D. N.;Hume. Determination of NTA in Environmental Water by Gas Chromatography, Anal. Lett., (1973), 6,' (41, 829-837.

Stolzberg, R. J., and D. N. Hume. Rapid Formation of Iminodiacetate from Photochemical Degradation of Fe (111) Nitrilotriacetate Solutions, Env. Sci. Technol., (1975), 2, 654.

Sturm R. N., and A. G. Payne. Environmental Testing of Trisodium Nitrilotri- acetate Bioassays for Aquatic Safety and Algal Stimulation, In Bioassay Techniques and Environmental Chemistry, Ed. Gary E. Glass, (1973), 403-424; 499 pp. Ann Arbor Science Publ. Inc., Ann Arbor, Michigan.

Swisher, R. D., M. M. Crutchfteld and D. W. Caldwell. Biodegradation of Nitrilotriacetate in Activated Sludge, Env. Sci. Technol., (1967), - 1, 820.

Tabatabai, M. A., and J. M. Bremner. Decomposition of Nitrilotriacetate (NTA) in Soils, Soil Biol. Biochem. , (1975), L, (2), 103-106.

Taylor, J. K., R. Alvarez, R. Paulson, T. C. Rains and H. L. Rook. Interaction of Nitrilotriacetic Acid with Suspended and Bottom Materials, Natl. Bureau of Standards, Washington, D.C. 24080, 31 pp. Water Polln. Control Res. Series 16020 GPR 07-71, Environmental Protection Agency (Progress Report) 1971.

Teratology Studies on Methylmercury ~~dr&ide and Nitrilotriacetate Sodium in Rats, Nature, (1973), 241, (5390: Feb: 16), 461-463.

Thayer S., and C. J. Kensler, Arthur D. Little, Inc. Current Status of the Environmental and Human Safety Aspects of Nitrilotriacetic Acid (NTA), Critical Reviews in Environmental Control, (1973), 2, (4), 375-404, Copyright, The Chemical Rubber Company, 1973.

Thom, N. S. Nitrilotriacetic Acid: A Literature Survey, Water Research, (1971), 5, (7), 391-399. -

Thompson,. . J. E., and J. R. Duthie. . The Biodegradability and. Treatability of NTA, J.W.P.C.F., (1968), 40, 305.

Thompson, S. W., R. A. Huseby, M. A..Fox, C. L. Davis and R. D. Hunt. Spon- taneous Tumors in the Sprague-Dawley Rat, Journal of the National Cancer Institute, (196.1), - 27, (5), 1037-1056.

Tiedje, J. M., and B. B. Mason. Biodegradation of Nitrilotriacetic Acid in Soils, Bacteriol. Proc., (1971), 71, 1.

. .

Tiedje, J. M., and B. B. Mason. Biodegradation of Nitrilotriacetate (NTA) in Soils, Soil Sci. Soc. Amer. Proc., (1974), 38, 278-283.

Tiedje, J. M., B. B. Mason, C. B. Warren and E. J. Malec. Metabolism of Nitrilotriacetate by Cells of Pseudomonas Species, Appl. Microbiol., (1973), - 25, (5), 811-818.

Tjalve, H. A Study of the Distribution and Teratogenicity of Nitrilotriacetic Acid (NTA) in Mice, Toxicol. Appl. Pharmacol., (1972), (2), 23, 216-221.

Trott, T., R. W. Henwood, and C. H. Langford. Sunlight Photochemistry of Ferric Nitrilotriacetate Complexes, Envtl. Sci. Technol., (1972), 6, (41, 367-368.

Usenik, P. M., and K. L. Murphy. Chlorination of Organic Chemicals, McMaster Univ., Hamilton, Ontario, Dept. of Chem. Eng. 73-1211-2, (1973).

Van Duuren, D. L., B. M. Goldschmidt, C. Katz, I. Seldman and J. S. Paul. Carcinogenic Activity of Alkylating Agents, J. Natl. Cancer Inst., (1974), 53, (31, 695.

Van Esch, G. J., and R. Kroes. The Induction of Renal Tumours by Feeding Basic Lead Acetate to Mice and Hampsters, Brit. J. Cancer, (1969), 23, 765.

Van Esch, G. J., H. van Genderen and H. H. Wink. The Induction of Renal Tumours by Feeding of Basic Lead Acetate to Rats,(1962), Brit. J. Cancer, 5, 289. - .

Walker, A. P. Ultimate Biodegradation of Nitrilotriacetate in the Presence of Heavy Metals, Progress in Water Technol., (1975); _Z, (3/4), 555-560.

Wallace, A., R. T. Mueller and G. V. Alexander. Effect of High Levels of Nitrilotriacetate on Metal Uptake by Plants Grown in Soil, Agron. J., (1974), 66, (5), 707-708. - Warren, C. B. (Monsanto Chemical Company). Reaction of NTA with Chlorine, July 1974 Report; December 1974 Report - unpublished.

Warren, C. B., and E. J. Malec. Biodegradation of Nitrilotriacetic Acid and Related Imino and Amino Acids in River Water, Science (Wash., D.C.), (1972), 176, (4032), 277-279. -

Warren, C. B., and E. J. Malec. Quantitative Determination of Nitrilotriacetic Acid and Related Aminopolycarboxylic Acids in Inland Waters: Analysis by Gas Chromatography, J. Chromatogr., (1972), 64, (2), 219-237.

Wong, P. T. S., D. Liu and B. J. Dutka. ~a~id'~iodegradation of NTA by a Novel Bacterial Mutant, Water Research, (1972), 5, 1577.

Wong, P. T. S., D. Liu and D. J. McGirr. Mechanism of NTA Degradation by a Bacterial Mutant, Water Research, (1973), z, 1367.

Woods, L. A., Chairman, J. Cornfield, R. A. Goyer, A. E. Martell, T. S. Miya, W. E. Ribelin, W. R. Richter, T. H. Shepherd and W. E. Sutton. Final Report: Assessment of the Potential of Nitrilotriacetate (NTA) to Compromise Human Health, Submitted by the Assistant Secretary for Health and Scientific Affairs' Ad Hoc Group on NTA, April 19, 1972.

Wright, P. L., M. L. Keplinger, I. D. Hill and J. C. Calandra. A Study of the Effect of Sodium Nitrilotriacetate on the Toxicity of Mercury Compounds, Toxicol. Appl. Pharmacol., (1972), 22, (2), 296.

Yang, S. -S. Thesis: Study of Na EDTA, University of Massachusetts, Amherst. Summary in Pd. Cosmet. Toxicol., ?1964), 2, 763-767.

~entsch, C. M., C. S. Yentsch, C. Owen and M. Salvaggio. Stimulatory ~ffects on Growth .and.Photosynthesis of the Toxic Red Tide Dinoflagellate, GonyauZax tamarmsis, with the ~ddition of Nitrilotriacetic Acid (NTA) , Environ. Lett., (19741, ;a, (3), .231-238.

. . . .

Zawirska, B., and K. Medras. Tumours and Disorders of the Porphyrin Metabolism in Rats with Chronic Experimental Lead'Poisoning, I. Morphologic Studies, Zbl. allg. Path. path. hat., (1968), E, 1.

Zitko, V., and W. V. Carson. Release of Heavy Metals from Sediments by Nitrilotriacetic Acid, Chemosphere, (1972), 2, 113.

Zollinger, H. U. Durch chronische Bleivergiftung erzeugte ~ierenadenome und - carcinome bei-Ratten und ihre Beziehungen zu den entsprechenden Neubildungen des Menschen, Virchows Arch. path. Anat., (1953), 323, 694.

APPENDIX II Task Force Membership

NTA TASK FORCE MEMBERSHIP

M r . Paul D. Foley Co-ordinator Development & Research Group P o l l u t i o n Control Branch Ontar io Min i s t ry of t h e Environment 135 St . C l a i r Avenue West Toronto, Ontar io M7V 1P5

D r . G. Becking Toxicology Divis ion Environmental Heal th D i r e c t o r a t e ~ u n n e y ' s Pas tu re Ottawa, Ontar io K I A OL2

D r . R . A. Goyer Chairman and Professor Dept. of Pathology Univers i ty of Western Ontar io Heal th Sciences Centre London, Ontar io N6A 5C1

D r . J. Xu l l e r Environmental Heal th Occupational Heal th P r o t e c t i o n Branch 15 Overlea Blvd. Toronto, Ontar io M4H 1A9

D r . H . L. Fa lk Assoc ia te D i r e c t o r f o r Heal th Hazard Assessment N.I.E.H.S. P. 0 . Box 12233 Research Tr i ang le Park, North Caro l ina 27709

D r . N . Chernoff Heal th E f f e c t s Research Lab. E.P.A. Research Tr i ang le Park, North Caro l ina 27711

D r . L . Golberg, M . B . , D.Sc., D.Phi1. ( ~ e c h n i c a l Advisor) P re s iden t Chemical Indus t ry I n s t i t u t e of Toxicology P. 0 . Box 12137 Research Tr i ang le Park, North Caro l ina 27709

D r . Michael Hogan Qechn ica l Advisor) Biometry Branch N.I.E.H.S. P. 0. Box 12233 Research Tr i ang le Park, North Caro l ina 27709

S e c r e t a r i a t R e s p o n s i b i l i t i e s

D r . A. E. P. Watson Research S c i e n t i s t I n t e r n a t i o n a l J o i n t Commission Great Lakes Regional Of f i ce 100 O u e l l e t t e Avenue,. 8 t h Floor Windsor, Ontar io N9A 6T3 (519) 256-7821 (313) 963-9041

Task Force Terms of Reference

TERMS OF REFERENCE FOR TASK FORCE ON ,THE HEALTH IMPLICATIONS OF NTA

INTRODUCTION

The data now in hand suggest strongly that phosphorus in effluents must be removed to the smallest concentration possible because some major sources, such as land runoff, are less controllable. Experience to date shows that phosphorus removal at some sewage treatment plants is less than had been expected and in addition creates significant sludge problems.

Replacements for P in detergents is apparently a more effective way to reduce P in STP effluent. It is probably a better environmental alternative from the standpoint of energy use and solid waste volume.

Canadian policy does not prohibit the use of NTA as P replacement but animal feeding studies in the U.S. suggesting possible cariinomas at very high doses have been used as a basis for not allowing the use of NTA in the U.S.

The RAB should provide advice to the IJC regarding a recommendation to Governments about the use of NTA. To do so, the RAB must review the NTA health related research.

CHARGE

A special task force needs to be created to:

(1) Bring together in an encapsulated form the research data pertaining to the health related research on NTA.

(2) Provide the RAB with advice as to the adequacy and validity of the research.

(3) Identify the health implications of the findings.

(4) Recommend a course of action for the International Joint Commission to take regarding the United States and Canadian policies on NTA.

The report of the task force should be delivered to the RAB during its first meeting of 1977.

Mr. Jim Amirsakis W. R. Grace Company Nashua, New Hampshire

Dr. Allan Bourquin U.S. EPA Environmental Research Lab. ,

Gulf Breeze, Florida

Mr. F. Alan Brownridge Manager, Prof. & Reg. Services Procter & Gamble Co. of Canada !

Hamilton, Ontario

Dr. Douglas G. Cowan Manager of Product Coordination Procter & Gamble International Div. Cincinnati, Ohio

Dr. Cypriano Cueto Carcinogenesis Program Bioassay Program National Cancer Institute Bethesda, Maryland

Dr. Gary Flamrn Assistant Director I

Division of Cancer Cause & Prevention, Bethesda, Maryland

Mr. Fred Huber W. R. Grace Company Nashua, New Hampshire

Mr. Robert R. Pollard Vice President, Marketing Organic Chemicals Division W. R. Grace Company Lexington, Massachusetts

Dr. Martin Rubin Consultant, W. R. Grace Co. Georgetown University

Dr. Lewis Scharpf, Jr. Manager, Research & Development Monsanto Industrial Chemicals Company St. Louis Missouri

Mr. George H. Scott Director, Corporate Professional & Reg. Services Research & Development Dept. Procter & Gamble, Cincinnati , Ohio

Dr. R. A. Squire, Formerly Acting Director Bioassay Program National Cancer Institute Bethesda, Maryland

Dr. Robert E. Winters Manager, Professional and Reg. Services Procter & Gamble Cincinnati, Ohio

Mr. J. W. Schmidt 1 Director, Wastewater Technology Centre Environmental Protection Service I

Canada Centre for Inland Waters 1 Burlington, Ontario ~

NCI Institute Data

INCIDENCE OF TUMORS I N MALE RATS (SRI, Na3NTA.H20)

Cont ro ls Low Dose Mid Dose High Dose 0 ppm 200 ppm 2,000 ppm 20,000 ppm

Inc idence of Urinary Tumors ( inc lud ing primary tumors of t he kidney, b ladder , and u r e t e r ) . 0124 (0%) 0123 (0%) 0124 (0%) 14/24 (38%)

S t a t i s t i c a l Tes tsa

95% Confidence I n t e r v a l s

P < 0.001, N.S. P < 0.001

Weeks t o F i r s t Observed Tumor --- --- --- 6 7

Inc idence of Tumors of t he Kidney 0/24 (0%) 0123 (0%) 0124 (0%) 9/24 (38%)


CI 95% Concidence I n t e r v a l s V

P < 0.001, N. S. N.S. P 0.001 P < 0.001 --- --- --- 3.41 - a:

Weeks t o F i r s t Observed Tumor --- --- --- 56

Inc idence of Trans i t iona l -Cel l Carcinoma of t h e Kidney



Weeks t o F i r s t Observed Tumor

N.S.

---

Inc idence of Tumors of t he Ureter 0124 (0%) 0123 (0%) 0124 (0%) 8/24 (33%)

S t a t i s t i c a l Testsa


N.S.

--- Weeks t o F i r s t Observed Tumor --- --- --- 5 6

N C I TABLE 2

INCIDENCE OF TUMORS I N FEMALE RATS (SRI, Na3NTA.H20)

Cont ro ls Low Dose Mid Dose High Dose 0 ppm 200 ppm 2,000 ppm 20,000 ppm

Inc idence of Primary Urinary Tumors ( i n c l u s i v e of pr imary tumors of t h e kidney, b l adde r , and u r e t e r ) 0124 (0%) 0124 (0%) 1/24 (4%) 13/24 (54%)

S t a t i s t i c a l ~ e s t s ~ N.S. P < 0.001

95% Confidence I n t e r v a l s --- --- --- 6.73 - a Weeks t o F i r s t Observed Tumor --- --- 99 4 2

Inc idence of Primary Tumors of t h e Kidney 0124 (0%) 0124 (0%) 0124 (0%) 4/24 (17%)

P S t a t i s t i c a l ~ e s t s ~

00


Weeks t o f i r s t Observed Tumor

P = 0.006, N.S. P 0.003

Inc idence of Tumors of t h e Ureter 0124 (0%) 0124 (0%) 0124 (0%) 6124 (25%)

S t a t i s t i c a l ~ e s t s ~


N.S.

---

N.S. P =, 0.011

--- 1.79 - a

Weeks t o F i r s t Observed Tumor --- --- --- . 42

Inc idence of Tumors of t h e Bladder 0122 (0%) 0123 (0%) 1/22 (5%) 5/24 (21%)

a S t a t i s t i c a l T e s t s


Weeks to First Observed Tumor

N.S.

N C I TABLE 3

INCIDENCE OF TUMORS I N MALE RATS ( L i t t o n , NTA)

Cont ro ls Low D ~ s e High Dose 0 PPm 7,500 ppm 15,000 ppm

-

Inc idence of Tumors of t h e Urinary T r a c t 0120 (0%) 1/49 (2%) 7/48 (15%)

S t a t i s t i c a l T e s t s a



P 0.021, N.S. P = 0.076 P = 0.006

a Beneath t h e un t r ea t ed c o n t r o l inc idence a r e two q u a n t i t i e s : ( a ) t h e P v a l u e f o r a 2x3 contingency t a b l e u s ing ch i squa re theory; (b) t h e P va lue f o r t h e t e s t f o r l i n e a r t r end i f a l i n e a r t r end f i t s t h e d a t a o r t h e word "nonlinear" , which means t h a t a l i n e a r t rend on a l o g i s t i c s c a l e does no t f i t t he d a t a .

Beneath t h e dosed group inc idence is t h e Fisher-Irwin one- ta i led t e s t f o r comparison of t h a t group wi th t h e c o n t r o l group when i t is below 0.10; o r , o therwise N.S. - n o t s i g n i f i c a n t .

This f o o t n o t e a p p l i e s t o N C I da t a l i s t e d i n Tables 1-9.

N C I TABLE 4

INCIDENCE OF TUMORS I N FEMALE RATS (LITTON, NTA)

Controls Low Dose High Dose 0 ppm 7,500 ppm 15,000 ppm

Incidence of Tumors of the Liver ( a l l non-malignant) 2/15 (13%) 8/49 (16%) 22/49 (45%)

S t a t i s t i c a l ~ e s t s ~ P = 0.003, N.S. p = 0.024 P - 0.001

95% Confidence In t e rva l s --- --- 1.00 - 52.13

Weeks t o F i r s t Observed Tumor 9 91104 104 9 3

Incidence of Tumors of the Urinary Tract 0120 (0%) 2/50 .(4%) 14/50 (28%)

a S t a t i s t i c a l Tests

95% Confidence In t e rva l s bl 0 Weeks t o F i r s t Observed Tumor

N.S. P = 0.005

--- 2.06 - a

Incidence of Tumors of the Bladder ( a l l malignant) 0118 (0%) 2/45 (43) 12/48 (25%)

S t a t i s t i c a l ~ e s t s ~

95% Confidence In t e rva l s

Weeks t o F i r s t Observed Tumor --- 104 91

Incidence of Tumors of the Lung ( a l l malignant ) 0115 (0%) 3/49 (6%) 7/46 (15%)

a S t a t i s t i c a l Tests

95% Confidence In t e rva l s

N.S. P - 0.033

N.S. . .

---

N.S.

---

Weeks t o F i r s t Observed Tumor --- 104 7 3

N C I TABLE 5

INCIDENCE OF TUMORS I N MALE MICE (LITTON, NTA)

Cont ro ls Low Dose High Dose 0 ppm 7,500 ppm 15,000 ppm

Inc idence of Tumors of t he Urinary Trac t ( a l l kidney tumors) 0120 (0%) 5/49 (10%) 24/44 (55%)

S t a t i s t i c a l ~ e s t s ~ P < 0.001, N.S. P < 0.001 nonl inear

95% Confidence I n t e r v a l s --- --- 6.34 - a Weeks t o F i r s t Observed Tumor 9 1 8 8 7 4

N C I TABLE 6

INCIDENCE OF TUMORS I N FEMALE MICE (LITTON, NTA)

Cont ro ls Low Dose High Dose 0 PPm 7,500 ppm 15,000 ppm

Inc idence of Tumors of t h e Urinary Trac t ( a l l malignant kidney tumors) 0120 (0%) 0139 (0%) 4/50 (8%)

S t a t i s t i c a l - T e s t s a



P = 0.086, N.S. P = 0.041

N.S.

. , . # . . . NCI TABLE 7

INCIDENCE OF TUMORS IN MALE MICE (LITTON, Na NTA.H20) 3

Controls Low Dose High Dose 0 PPm 2,500 ppm 5,000 ppm

Incidence of Tumors of the Hematopoietic System (a11 malignant) 0120 (021 4/47 (9%) 9/50 (18%)

Statistical ~ e s t s ~ N.S. P = 0.038

95% Confidence Intervals - - - . --- 1.11 - a Weeks to First Observed Tumor --- 7 5 8 6

NCI TABLE 8

SUMMARY OF PRIMARY EPITHELIAL NEOPLASMS OF THE URINARY TRACT - RATS (mM NTA/kg Diet)

I All Controls

KIDNEY

( Transitional - Cell Papilloma 1 0163 0164 1 / Tubular-Cell Adenocarcinoma 1 0163 0164 1

Tubular-Cell Adenoma

URETER Transitional<ellCarcinoma

Papillary Adenoma ,0164 0164

URINARY BLADDER Transitional-Cell Carcinoma 1 0157 0156 1

I Squamous-Cell Carcinoma ( 0157 0156 1 Total Number of Animals with

These Urinary Tract Tumors 0 0

* In female rat receiving 2.0.00 ppm.

Na NTA-H20 3 Na3 NTA-H 0

7 mM1kg 27 mM1kg 2,000 ppm 7,500 ppm

M/F l M 2F

NTA

40 mM1kg 7,500 pprn M F

Na3 NTA-H20

54 mM1kg 15,000 pprn M F

Na3 NTA-H20

70 mM/kg 20,000 pprn M F

NTA

80 mM/kg 15,000 pprn M F

NCI TABLE 9 ..: .

S-Y OF PRIMARY EPITHELIAL NEOPLASMS OF THE URINARY TRACT - MICE

A1 1

Controls

Na3NTA.H20 Na 3NTA. H20 NT A NTA

9 mM/kg 18 mM/kg 40 mM/kg 80 m~/kg

2,500 ppm 5,000 ppm 7,500 ppm 15,000 ppm

SPECIFIC ORGAN/TUMOR TYPE M F M F M F M F M. F VI CI

KIDNEY

Tubular-Cell Adenoma 0/40* 0139 - - - - - - 1/43(2%) -

Tubular-Cell Adenocarcinoma 0140 0139 - - - - 5/48(10%) - 22/43 (50%) 4/30(8%)

- - - - - - 1/43(2%) - 0140 0139 Papilloma

Total Number of Animals with 0 0 0 0 0 0 These Urinary Tract Tumors

"~enominators equal number of tissues examined histopathologically.

IJC Great Lakes Research Advisory Board

Terms of Reference

GREAT LAKES RESEARCH ADVISORY BOARD TERMS OF REFERENCE

1. As used herein, "research" includes development, demonstration and research activities, but does not include regular monitoring and surveillance of water quality.

2. The functions and responsibilities of the Research Advisory Board relating to research activities in Canada and the United States concerning the quality of the waters of the Great Lakes System shall be as follows:

(a) To review at regular intervals these research activities in order to:

(i) examine the adequacy and reliability of research results, their dissemination, and the effectiveness of their application;

(ii) identify deficiencies in their scope, and inadequacies in their funding and in completing schedules;

(iii) identify additional research projects that should be undertaken;' (iv) identify specific research programs for which international

cooperation will be productive;

(b) To provide advice and consolidations of scientific opinion to .. the Commission and its boards on particular problems referred

to the Advisory Board by the Commission or its boards;

(c) To facilitate both formal and informal international cooperation and coordination of research;

(d) To make recommendations to the Commission.

3. The Research Advisory Board on its own authority may seek analyses, assessments and recommendations from other professional, academic, governmental or intergovernmental groups about the problems of the Great Lakes water quality research and related research activities.

4. The International Joint Commission shall determine the size and composition of the Research Advisory Board. The Commission should appoint members to the Advisory Board from appropriate Federal, State and Provincial Government agencies and from other agencies, organizations and institutions involved in Great Lakes research activities. In making these appointments the Commission should consider individuals from the academic, scientific and industrial communities and the general public. Membership should be based primarily upon an individual's qualifications and potential contribution to the work of the Advisory Board.

5 . The Research Advisory Board should work at all times inclose cooperation with the Great Lakes Quality Board.

IJC Great Lakes Research Advisory Board

Membership L i s t


UNITED STATES SECTION

I Dr. Donald I. Mount (Chairman) Director Environmental Research Laboratory -

Duluth 6201 Congdon Avenue Duluth, Minnesota 55804 (218) 727-6692 FTS 783-9548

Professor Joseph Shapiro Geology and Ecology Department University of Minnesota 220 Pillsbury Hall Minneapolis, Minnesota 55455

Dr. Eugene J . Aubert Director Great Lakes Environmental Research

Laboratory National Oceanographic and

Atmospheric Administration 2300 Washtenaw Avenue Ann Arbor, Michigan 48104 (313) 994-3374

Professor Archie J. McDonnell Department of Civil Engineering Water Resources Research Center The Pennsylvania State University University Park, Pennsylvania 16802 (814) 865-9362 (814) 863-0291

Dr. Virginia L. Prentice Associate Research Geographer Environmental Research Institute of

Michigan P. 0. Box 618 Ann Arbor, Michigan 48107 (313) 994-1200

Dr. Mitchell R. Zavon Corporate Medical Director Hooker Chemicals and Plastics Corp. 222 Rainbow Blvd. North P. 0. Box 728 Niagara Falls, New York 14302 (716) 278-7408

Dr. Herbert E. Allen Associate Professor Dept. of Environmental Eng. Illinois Institute of Technology Chicago, Illinois 60616 (312) 567-3559

Mr. John J. Convery Director Wastewater Research Division Municipal Environmental Research Lab. U.S. EnvironmentalProtection Agency Cincinnati, Ohio 45268 FTS: 684-7601

Secretariat Responsibilities

Dr. Dennis E. Konasewich Research Scientist Great Lakes Regional Office International Joint Commission 100 Ouellette Avenue, 8th Floor Windsor, Ontario N9A 6T3 (519) 256-7821 (313) 963-9041

GREAT LAKES 'RESEARCH ADVISORY BOARD

CANADIAN SECTION

D r . A. R. LeFeuvre (Chairman) D i r e c t o r Canada Centre f o r Inland Waters Environment Canada P. 0 . Box 5050 Burl ington, Ontario L7R 4A6 (416) 637-4272

M r . Paul D. Foley Coordinator , Development & Research Group

' P o l l u t i o n Control Branch 'Ontar io Minis t ry of t h e Environment 135 S t . C l a i r Avenue West Toronto, Ontario M4V 1P5 (416) 248-3739

M r . J. Douglas Roseborough, D i rec to r F i sh and W i l d l i f e Research Branch Ontar io Minis t ry of Natura l Resources P. 0 . Box 50 Maple, Ontar io LOJ 1EO (416) 832-2261

D r . J . R. Val lentyne Senior S c i e n t i f i c Advisor Ocean & Aquatic A f f a i r s F i s h e r i e s & Marine Serv ice Environment Canada 580 Booth S t . Ottawa, Ontar io K I A OH3 (613) 992-8989

Mrs. F. Edna Gardner 110 North Drive I s l i n g t o n , Ontar io M9A 4L2 (416) 231-3418

D r . G. H. Tomlinson, I1 Vice Pres ident Research & Development Technology Domtar Limited P. 0. Box 7210 Montreal, Quebec' H3C 3M1 (514) 282-5400

D r . . James H. Day Divis ion of Allergy Kingston General Hospi ta l Kingston, Ontario K7L 2V7 (613) 547-3309

Prof. Jose Llamas acuity of Science & Eng. Water Resources Center P a v i l l i o n Pou l io t Univers i ty of Lava1 Rue 3713 S t . Foy, Quebec J 1 K 7P4

S e c r e t a r i a t R e s p o n s i b i l i t i e s

D r . Dennis E . Konasewich Research S c i e n t i s t Great Lakes Regional Of f i ce I n t e r n a t i o n a l J o i n t Commission 100 O u e l l e t t e Avenue, 8 t h F l o o r Windsor, Ontario N9A 6T3 (519) 256-7821 (Windsor) (313) 963-9041 (De t ro i t )


EX OFFICIO MEMBER -

Great Lakes Fishery Commission and IAGLR (President)

Carlos M. Fetterolf, Jr. Executive Secretary Great Lakes Fishery Commission 1451 Green Road Ann Arbor, Michigan 48107 (313) 662-3209 (FTS) 374-5481

research awi! jory board - international joint … · following report on its findings prepared by...

Documents