TOXICITY OF ORGANIC CHEMICALS TO EMBRYO-LARVAL STAGES OF FISH potx

EPA-560/11-7~-007·

TOXICITY OF ORGANIC CHEMICALS

TO EMBRYO-LARVAL STAGES OF FISH

June 1979

Final Report

Contract No. 68-01-4321

Wesley J. Birge

Jeffrey A. Black

Donald M. Bruser

Project Off; cer

Arthur M. Stern

U.S. Environmental Protection Agency

Office of Toxic Substances

Washington, D.C. 20460

PBPO-10I637

'-'~

. TECHNICAL REPORT DATA (pt_l'IltI.tll~tiImJ or. the TPUJ. btt/Dnl t:ompll1tiJf8J

1. RepORT iIIO. 12. :'3. RECIPIENT'S ACCES$IONroNO.

EPA-560/11-79-007 PI:) Y() - /f!) /c.. 3 '7

4. TITLE ANO SUBTITLE I. REPORT DATE I

Toxicity of Organic Chemicals to Embryo-Larval June 1979 (Date of Issue)

$tages of Fish lJ. PEFlIIOAMINQ OFlGANlZATION CODe

7. AUTHOAlSJ ~. PERFOAMING ORGANIZATION REPOAT NC .

.. Wes1ey J. Bi rge, Jeffrey A• Black, and Donald M. Bruser

• I. PERFORMING ORGANIZATION NAME AND AOOFlES$ 10. PROGRAM ELeMENT NO•

rhomas Hunt Morgan School of Biological Sciences

University of Kentucky . 11. C1JNTftA~/(iI'lANTNO.

~ex1ngton, KentuckY 40506 . 68..01-4321 . 12. SPONSOAING AGENCY NAME AND AOORESS 13. TYPE OF REPORT AND PERIOD COVERED

pff1ce of Toxic Substances Final (Oct. 1976-Feb. 1979) ~~S. Environmental ProteCtion Agency , ..SPONSORING AGENCY cooe

~ashfngton, D.C. 20460 . . 1•• SUPPL&MENTAAY NOTES

.

,

1lJ.AATftACTA continuous flow procedure was developed for evaluating effects of insoluble

and volatile organics on embryo-larv&l stages of fish. Test compounds were selected

for different combinations of'solubility and volatility and included aniline, atrazine, chlorobenzene, chloroform, 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid, d10ctyl ~hthalate, malathion, trisodium nitrilotriacetic acid, phenol, and polychlorinated bi￾phenyl (Capacitor 21). Aclosed system devoid of standing air space greatly reduced

volatility as a test variable. Mechanical homogenization proved highly effective in

suspending hydrophobic compounds in influent water. Continuous agitation in the test

chamber\nd regulation of detention time further'precluded the need for carrier sol￾vents. Test results indicated good reproducibility of exposure concentrations. The

most toxic compounds included Capacitor 21, chlorobenzene, 2,4-dichlorophenol, and

phenol: Chlorobenzene at 90 pg/l produced complete lethality of trout eggs. The three

other compounds gave log probit LCSO's of 2 to 70 ug/l when trout stages were exposed in

hard water, and LCII,swere 0.3, 1.0, and 1.7 pg/l for phenol t Capacitor 21, and 2,4~di￾chlorophenol. Chloroform also was· highly toxic to trout stages and Lells ranged from

4.9 to 6.2 Ug/1. -When bass and goldfish stages w~re exposed to chlorobenzene, LCl's

ranged from 8 to 33 pg/l. Compared to other species, trout developmental stages gener- ally exhibited the greatest sensitivity. The LCI values determined in embryo-larval

tests compared closely with maximum acceptable toxicant concentrations developed in I Hf,,_rvrlA dwH..tIl! "'net' ~2hlD ies nf teratic larvae . 17. KEY WORDS AND DOCUMENT ANAl.YSIS

iI. oeSCFlU"TOFlS b.IOENTIFIEFlS/OPEN ENDED TEAMS Co (;OSATI Field/G:oup

Toxicity Tests - Embryos

Larvae .- Embryonic Lethality

Freshwater Fish larval Lethality

Bioassay Teratogenesis

Terata Organic ,Toxicants

Organic Compounds Volatile Organics Insolubl~ ~rganics Water Quality ICarr;-r SolYe~ts i

1a. DISTRIBUTION STATEMENT 19.5ECUFUTY CLASS {nli:l Rl1pOITJ 21. NO. OF PAGiiS I

Unclassified '12. Release Unlimited 20. SECURITY Cl...\SS (Thu pap) 22.l"RICEA:? A1J tt- Unclassified I /)1F-HOl

NOTICE

This report has been reviewed by the Office of

Toxic Substances, EPA, and approved for publication.

Approval does not signify that the contents nec~ssar1ly

reflect the views and policies of the Environmental

Protection Agency, nor does mention of trade names or

commercial products constitute indorsement or

recommendation for use.

;1

ABSTRACT

A continuous flow procedure was developed for evaluating effects of

insoluble and volatile organics on embryo-larval stages of fish. A closed system

devoid of standing air space was used to minimize volatility as a test variable.

!nsoluble compounds were suspended in influent water by mechanical homogeniza￾tion_ without the use of carrier solvents. Tests were performed on aniline,

atrazlne, chlorobenzene, chloroform, 2,4-dichlorophenol, 2,4-dichlorophenoxy￾acetic acid (2,4-0), dioctyl phthalate (OOP), malathion, trisodium nftrllo￾triacetlc acid (NTA), phenol, and polychlorinated biphenyl (Capacitor 21).

Maintaining water hardness at 50 and 200 mg/l CaC03, exposure was continuous

from fertilization through 4 days posthatching for largemouth bass, bluegill

sunfish, channel catfish, goldfish, rainbow trout, and redear sunfish.

Exposure levels which produced 50% (LeSO) and 1% {LeI} control-adjusted

impairment (lethality, teratogenesis) of test populations were calculated by

log probit analysis. The LCI's were used as a basis for estimating threshold

concentrations for toxic effects. To determine reliability of LCI values, they

were compared with maximum acceptable toxicant concentrations (MATC) developed

in partial and complete life-cycle studies. Good correlations were obtained

when data were adequate to permit comparisons, and the findings indicated that

LCI values determined in embryo-larval tests carried through 4 days posthatching

were useful in estimating long-term @-jrfects of aquatic pollutants.

Test results indicated good reprQducibility of exposure concentrations

for both volatile and insoluble toxicants. The most toxic compounds included

Capacitor 21, chlorobenzene, 2,4-dichlorophenol, and phenol. Chlorobenzene at

90 pg/l produced complete lethality of trout eggs, and LCI's ranged from

8 to 33 pg/l in tests with the largemouth bass and goldfish. The tnree

other compounds gave log probit lCSO'S of 2 to 70 pg/l when trout stages were

exposed 1n hard water, and LeI's were 0.3, 1.0, and 1.7 pgll for phenol,

Capacitor 21, and 2,4-dichlorophe~01. Phenol was less toxic to developmental

stages of the goldfish and bluegill. When tests were conducted in hard water,

the LCSO'S were 0.34 and 1.69 mg/l and the LC1's varied from 2.0 to 8.8 ~g/l.

Depending on water hardness, LCI's determined in P9/l with the rainbow trout

iii

ranged from 4.9 to 6.2 for chloroform, 21.9 to 32.5 for 2,4-0, and 29.0 to

77.2 for atraztne. Though not tested on the trout, LCt'S determined with the

goldfish ranged from 143.2 to 215.0 ~g/l for aniline and 141.1 to 439.6 ~g/l

for malathion. The organics least toxic to the trout included NTA and COP. and

the LCSO's varied from 90.S to 114.0 and 139.5 to 149.2 mg/l, respectively.

Compared to the other species, trout developmental stages generally exhibited

the greatest overall sensitivity. Though water hardness did not substantially

alter toxicity of the selected organic compounds. phenol was somewhat more

toxic 1n hard water. All compounds produced appreciable frequencies of teratic

larvae.

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TABLE OF CONTENTS

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Teratogenesis in Fish Embryos .

SUMfttA.RY • • • • • • • •• " • •

REFERENCES .• . . . • • • • . . . • . •

ABSTRACT .••

LIST OF TABLES ••.••••••.••

LIST OF FIGURES • • • • . • . •

ACKNOWLEDGMENTS .

INTRODUCTION .••

CONCLUSIONS • • . • • • •

RECOMMENDATIONS • • • • • • • • • .

DEVELOPMENT OF TEST SYSTEM AND PROCEDURES • .

Materials and Methods ••••••••••

Selection of animal species • • • . • . • •

Selection of organic toxicants ••••••.

Test conditions and expression of data

Test water • • • . • • • • • . • • • •

Embryo-larval test system .•••••

Analytical procedures •••••

Initial Performance Evaluation

APPLICATION OF TEST SYSTEM • . .

Embryo-Larval Toxicity Tests

Aniline • • • • • •• Atrazine • • • • • • . • • •

Capacitor 21 • • • .

Chlorobenzene • •...

Chloroform • • • • • • • . • •

2,4-Dichlorophenol •.•••••

2,4-Dichlorophenoxyacetic acid

Dloctyl phthalate ••.•.•••

Malathion •••••••••••••.•

Trisodium nitrilotriacetic acid ••••••• Pheno1 ••••••••••

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LIST OF TABLES

Table

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49

52

54

50

51

48

11

13

23

38

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• . • . 55

• • • . 22

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Organic compounds used 1n toxicity tests •••••••••

Toxicity tests performed on embryo-larval stages of fish

Water quality characteristics observed

during toxicity tests with organic compounds

Reconstituted test water • • •.•••...••

Regulation of Sudan IV-chlorobenzene

1n continuous flow tests '!'"

Regulation of organic compounds 1n continuous flow

toxicity tests with fish embryo-larval stages

Log problt LCSO values for organic compounds

Log probit LCI values determined at

4 days posthatchlng for organic compounds . • • • • • 41

Toxicity of aniline to embryo-larval stages of fish 43

Toxicity of atrazine to embryo-larval stages of fish 44

Toxicity of Capacitor 21 to embryo-larval stages of fish •••• 45

Toxicity of chlorobenzene to embryo-larval stages of fish • • . • 46

Toxicity of chloroform to embryo-larval stages of rainbow trout. 47

Toxicity of 2,4-dichlorophenol (DCP)

to embryo-larval stages of fish • • • • • •.•.•

Toxicity of 2,4-dichlorophenoxyacetic acid (2,4-0)

to embryo-larval stages of fish .

Toxicity of dioctyl phthalate (DOP)

to embryo-larval stages of fish.

Toxicity of malathion to embryo-larval stages of goldfish ••

Toxicity of trisodium nitrilotriacetic acid (NTA)

to embryo-larval stages of fish ••••••••..

Toxicity of phenol to embryo-larval stages of fish

Comparison of LeI's determined in embryo-lar~~:

tests wi th MATt I S der; ved from 1ife-eye1e s tu'~ I es

7

8

1

2

3

4

5

6

19

20

17

18

15

16

9

10

11

12

13

14

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