INVESTIGATIONS IN FISH CONTROL

98. History of Acute Toxicity Tests with Fish, 1863-1987

UNITED STATES DEPARTMENT OF THE INTERIOR 

FISH AND WILDLIFE SERVICE
Investigations in Fish Control, published by the Fish and Wildlife Service, include reports on the results of work 

at the National Fisheries Research Center at LaCrosse, Wisconsin, and reports of other studies related to fish control 

and acute toxicity of piscicides. Though each report is regarded as a separate publication, several may be issued 

under a single cover, for economy. See inside back cover for list of current issues.

Copies of this publication may be obtained from the Publications Unit, U.S. Fish and Wildlife Service, 18th 

and C Streets, N. W., Mail Stop 1111, Arlington Square Building, Washington, DC 20240, or may be purchased 

from the National Technical Information Service (NTIS), 5285 Port Royal Road, Springfield, VA 22161.

Library of Congress Cataloging-in-Publication Data

Hunn, Joseph B.

History of acute toxicity tests with fish, 1863-1987 / by Joseph 

B. Hunn.

p. cm. (Investigations in fish control ; 98)

Supt. of Docs, no.: I 49.70:98

1. Fishes Effect of water pollution on Testing. 2. Water 

quality bioassay. 3. Toxicity testing. I. Title. II. Series. 

SH157.7.I58 no. 98 

[SH174]

639.9'77 s dc!9 89-600014 

[597'.2'4] CIP
INVESTIGATIONS IN FISH CONTROL

98. History of Acute Toxicity Tests with Fish, 1863-1987

By Joseph B. Hunn

U.S. Fish and Wildlife Service 

National Fisheries Research Center LaCrosse

P.O. Box 818 

LaCrosse, Wisconsin 54602

1989

History of Acute Toxicity Tests with Fish, 1863-1987

by

Joseph B. Hunn

U.S. Fish and Wildlife Service 

National Fisheries Contaminant Research Center

Route 2 

Columbia, Missouri 65201

Abstract

Acute toxicity tests with fish were first reported in 1863. Test methodology developed slowly 

before World War n, during the period when the goldfish (Carassius auratus) was the recommended 

test animal. After the war, considerable effort was made to standardize test methods. A standardized method was published in 1960, refined in 1975, and updated in 1980. The primary use of the 

static acute toxicity test is to determine the median concentrations of chemicals that are lethal to 

aquatic organisms. Other applications include establishing the half-life of the biological activity of 

chemicals, evaluating the toxicity of mixtures of chemicals, and developing quantitative structureactivity relations used in predicting the toxicity of particular groups of chemicals. Data from acute 

toxicity tests are now required for the registration of chemicals for use in fishery work by the U.S. 

Food and Drug Administration and the Environmental Protection Agency. Acute toxicity data are 

also used in ecological risk assessment. Static acute toxicity tests will continue to play an essential 

role in aquatic toxicology in the foreseeable future.

Although the history of water pollution research was 

discussed by Hynes (1960), Warren (1971), and Tarzwell 

(1978), only limited attention has been given to the use 

offish in acute toxicity tests. Since little historical background is given in recent texts on aquatic toxicology 

(Nriagu 1983; Rand and Petrocelli 1985), the present 

review is intended to be an introduction to toxicity testing 

or of interest to those seeking background information on 

acute toxicity tests. Certain persons and events are highlighted because of their importance in the development 

of test techniques or contributions to the evolution of acute 

toxicity testing. Toxicity testing with fish is briefly summarized later, in the Table.

Statistical procedures used in evaluating aquatic toxicity 

tests are not discussed. Statistical approaches used to 

estimate toxicity in acute tests were given by Brown 

(1973), Stephan (1977), and Gelber et al. (1985).

In much of the early literature on pollution studies with 

fish, the term "bioassay" described what is now termed 

a "toxicity test." These two terms, as defined by Rand 

and Petrocelli (1985), are now distinct: A bioassay is used

to evaluate the relative potency of a chemical by comparing its effect on a living organism with the effect of a 

standard preparation (e.g., a reference hormone or toxicant) on the same type of organism; a toxicity test is the 

means used to determine the toxicity of a chemical or other 

test material.

The Early Years, 1863-1939

Fishes have been used in experiments for nearly 

200 years. Early research included studies of experimental 

embryology and parasitology, genetics, nutrition, renal 

physiology, endocrinology, and nerve physiology (Nigrelli 

1953). One of the first reports of the use of fish in toxicity studies was that of Penny and Adams (1863). 

Laboratory tests were performed on the effects of 

chemicals (used in dye-works) to two fishes a minnow 

because it was sensitive to "disturbing influences" and 

the goldfish (Carassius auratus) because of its "tenacity 

for life.'' Penny and Adams conducted more than 400 ex1
periments with 71 chemicals. Water for testing came from 

the River Leven, and toxicants were added in the laboratory. Powers (1917) published a monograph proposing 

the goldfish as a test animal in toxicity studies and introduced the concept of measuring survival time versus 

concentration of the test chemical. This basic concept is 

still used in some applications such as time to 50% mortality and lethal threshold concentration (Sprague 1973).

The effect of the mining of lead (Pb) on stream fauna 

and flora was studied in England during the 1920's. 

Carpenter (1925) was one of the first investigators to use 

caged fishes to determine the toxicity of runoff from mine 

tailings and to conduct laboratory tests with river water 

to examine the toxicity of Pb to the minnow Leuciscus 

phoxinus and to goldfish. Parallel laboratory studies were 

also conducted at concentrations similar to those measured 

in the field (Carpenter 1925). Signs of Pb poisoning, such 

as increased mucus production and decreased survival 

time at those Pb concentrations, supported the inference 

that the mortality observed in the field was due to Pb 

poisoning. Additional studies with 13 other species of 

freshwater fishes showed that the exposure of fish to Pb 

salts in distilled water caused similar signs of intoxication (Carpenter 1930). Carpenter also noted that the introduction of small amounts of hard water into test vessels 

containing distilled water as the test medium caused erratic 

results in Pb toxicity tests.

Belding (1927) provided an excellent early critical 

review of conditions necessary for the use of fish in the 

successful testing of polluting substances. He identified 

important variables that concerned the test animal and 

dilution water. The important test variables related to fish 

included: individual variation; differences among families 

or species of fishes; differences in age, size, weight, and 

vitality; and differences in the environment (water quality, 

number offish per container, dissolved oxygen, temperature). Six characteristic signs of poisoning irritation, 

inactivity, erratic swimming behavior, oxygen hunger, 

loss of equilibrium, and increased or decreased respiration rate were suggested for use in documenting the toxicity of the test substances. Steinmann (1928) was the first 

to measure both heart rates and respiration rates of fish 

exposed to chemicals. Interest in relating biochemical and 

physiological responses to chemical exposure later developed into sophisticated approaches for defining acute toxicity syndromes in fish (McKim et al. 1987).

The first detailed investigation that combined field and 

laboratory studies on the toxicity of pollutants to fishes 

was conducted by Ellis (1937). His work was notable for 

three reasons: (1) he conducted extensive field studies to 

define water quality characteristics associated with healthy

aquatic communities, including some recommended water 

quality values such as dissolved oxygen concentration and 

pH range that remain applicable today; (2) he compiled 

the known modes of action and toxicity of 114 toxic 

substances; and (3) he and his colleagues performed many 

laboratory toxicity tests with goldfish and Daphnia magna 

in soft and hard waters. He also established hazard rankings to fishes (none, possible, moderate, and critical) for 

30 common types of municipal and industrial effluents.

Many early investigators used distilled water as the test 

medium to reduce possible contamination with metals; 

however, its use was later found to impart an additional 

osmotic stress to freshwater fishes (Hunn and Alien 1964). 

In experiments with the threespine stickleback (Gasterosteus aculeatus), Jones (1938) noted that Pb was far less 

toxic in hard tap water containing 50 mg/L of calcium 

than in soft tap water containing only 1 mg/L. Further 

studies with test waters containing calcium that ranged 

from 0 to 50 mg/L (as calcium chloride or calcium nitrate) 

documented the ameliorating effects of calcium on Pb toxicity, which included reductions in respiratory distress and 

in the accumulation of coagulated mucus. Later studies 

with goldfish and with mucus from the eel Anguilla 

anguilla showed that excess coagulated mucus was not 

formed in the presence of sufficient calcium; however, 

if Pb first interacted with the mucus to cause coagulation, 

the addition of calcium did not reverse the process, 

because Pb has a greater affinity than calcium for binding sites on mucus.

Early observations by Carpenter (1930) and Jones 

(1938) on the effects of hardness or calcium on the toxicity of divalent metals ultimately led to water hardness 

criteria for certain metals. For example, criteria for cadmium and lead are based on hardness; lower allowable 

concentrations were suggested for soft water man for hard 

water (Thurston et al. 1979).

The Middle Years, 1940-1960

A new era in aquatic toxicology in the 1940's started 

with the advent of organochlorine insecticides (Dunlap 

1981). To study problems created by these compounds 

and other pollutants in fresh water, the U.S. Public Health 

Service established the Aquatic Biology Section at its 

laboratory at Cincinnati, Ohio, in 1948 (Tarzwell 1978). 

The next 10 years introduced the era of the "pickle-jar 

biologist," as the 5-gallon pickle jar became the standard 

vessel for acute toxicity tests (Fig. 1). Accompanying the 

expansion of research into pollution problems and the need 

to establish water quality criteria, two extensive critical
'ir

Fig. 1. Making observations during a static 

acute toxicity test in the widely-used 5-gallon 

pickle jars of 1940-60 at the Fish Control 

Laboratory, LaCrosse, Wisconsin. Photo by 

W. Mauck.

reviews of the toxicity of industrial wastes and their components to fish were published by Doudoroff and Katz 

(1950,1953). During this period numerous toxicity studies 

with freshwater fishes were being conducted at the U.S. 

Fish and Wildlife Service Laboratory, Leetown, West 

Virginia (Wood 1953; Hollis and Lennon 1954). More 

than 4,000 chemicals were screened for their toxicity to 

six species of fish: brook trout, Salvelinus fontinalis; 

brown trout, Salmo trutta; rainbow trout, 5. gairdneri; 

bluegill, Lepomis macrochirus; yellow perch, Perca 

flavescens; and goldfish.

Another large-scale toxicity testing program began in 

1953 at the Hammond Bay Biological Station of the U.S. 

Fish and Wildlife Service, in a search for a selective toxicant to control the sea lamprey (Petromyzon marinus) in 

the Great Lakes. The testing of more than 4,000 chemicals 

led to the discovery of the selective larval lampricide 

3-trifluoromethyl-4-nitrophenol (Applegate et al. 1957; 

Howell et al. 1980).

Soon after World War n, a major element in the development of toxicity testing was a drive to standardize test 

methods. One early attempt (Hart et al. 1945) was limited 

in scope and effect because only 300 copies of a manual 

of suggested procedures were published. After the manual 

was published, a committee under the leadership of Peter 

Doudoroff was established by the Committee on Research 

of the Federation of Sewage and Industrial Wastes 

Association now the Water Pollution Control Federation (Doudoroff 1986). This committee gathered materials 

from many people conducting toxicity studies with fish 

and aquatic invertebrates (Doudoroff et al. 1951). The 

report laid the foundation for the acceptance and later 

publication of standardized test procedures for the first 

time in the 11th edition of Standard Methods for the Examination of Water and Wastewater (American Public 

Health Association et al. 1960).

By the late 1950's, the number of agencies conducting 

aquatic lexicological research had increased substantially.
Fig. 2. The U.S. Fish and Wildlife Service Fish Control Laboratory at the junction of the Black, LaCrosse, and Mississippi 

rivers was operated in 1959-78, primarily to develop methods for the control of rough fish.

For example, two Fish and Wildlife Service laboratories 

were established in 1959: the Fish Control Laboratory at 

LaCrosse, Wisconsin (Fig. 2), under the direction of 

Robert E. Lennon, and the Fish-Pesticide Laboratory at 

Denver, Colorado, under the direction of Oliver B. Cope. 

In 1966, the Fish-Pesticide Laboratory was transferred 

to Columbia, Missouri. Simultaneously, the capacity of 

the Biology Section of the U.S. Public Health Service 

laboratory in Cincinnati to conduct toxicity tests was greatly expanded at the Newtown (Ohio) State Fish Hatchery. 

Greatly expanded research facilities and progress in 

analytical chemistry markedly increased scientists' abilities to detect and measure pollutants and test their toxicity 

to fish. The development of improved and sophisticated 

spectrophotometric methods of analysis, including atomic 

absorption spectroscopy and gas-liquid partition chromatography by A. T. James and A. J. P. Martin (1952, as 

cited by Ettre 1975), enabled investigators to measure test 

concentrations of chemicals. Although actual concentrations of many chemicals could be measured, numerous 

studies continued to be conducted in which only calculated 

(nominal) concentrations were used (Mayer and Ellersieck 

1986).

The Recent Years, 1961-1987

Standardization of test procedures in aquatic toxicity 

testing has been a primary goal of toxicologists in recent 

years. The Committee on Methods for Toxicity Tests with 

Aquatic Organisms (1975), represented by the Environmental Protection Agency, U.S. Fish and Wildlife Service, and private industry, first recognized the importance 

of common methods among all researchers involved in 

aquatic toxicity tests. This committee recommended 

common species, test temperatures, test waters, and procedures for toxicity tests. Reconstituted test water formulations and pH buffer schedules developed at the Fish
Control Laboratory, LaCrosse, Wisconsin (Marking 

1969), were adopted by the committee. The standardization work of this committee was further promoted by the 

American Society for Testing and Materials (ASTM). The 

work of the 1975 committee was adopted with only format changes by ASTM and published as the standard practice for conducting acute toxicity tests with fishes in 1980. 

Then, in the 15th (1981) edition of Standard Methods for 

Examination of Water and Wastewater, the American 

Public Health Association, American Water Works 

Association, and the Water Pollution Control Federation 

adopted many of the standards promoted by the 1975 committee, including the use of reconstituted water formulations for standardized tests. Thus, the late 1950's dream 

of a standardized methodology for acute toxicity tests was 

fulfilled.

Throughout this period, attempts have been made to 

better define the biotic and abiotic factors that affect the 

outcome of toxicity tests. Biotic characteristics include 

such variables as test species, life stage, nutrition, health 

of test organisms, and acclimation; abiotic factors include 

temperature, hardness, alkalinity, and pH (Sprague 1985). 

Mayer and Ellersieck (1986), in their analysis of more 

than 4,000 toxicity tests with 410 chemicals (mostly 

organic pesticides) and 66 species of aquatic animals, 

found an increase or decrease in LC50 (the concentration estimated to be lethal to 50% of the test organisms after 96 h of exposure) per 10° C change in 

temperature similar to that predicted by the Qio concept. 

However, hardness had little effect on the toxicity of 

organic chemicals that could not be explained by changes 

due to pH, even though hardness ions especially 

calcium sometimes greatly influence the toxicity of 

metals to fish (Hunn 1985; Mance 1987), as first 

documented by Jones (1938). Although pH affected toxicity in only 20% of the chemicals examined, it caused 

the greatest average change in toxicity (Mayer and Ellersieck 1986).

Hydrogen ion concentration (pH) influences chemicals 

in at least three ways. (1) It can change the physical state 

of the molecule (ionization), which in turn changes its 

lipid-solubility and thus its bioavailability (Hunn and Alien 

1974; Marking 1975). Although changes in lipid-solubility 

can explain much of the change in toxicity of some 

chemicals such as 3-trifluoromethyl-4-nitrophenol (Marking and Olson 1975), the uptake and toxicity of ionized 

molecules have not been fully determined and the 

mechanism of uptake remains speculative (Alien and Hunn 

1986). (2) The pH can influence the rate of degradation 

(hydrolysis) of the chemical (Mayer and Ellersieck 1986). 

(3) The pH also affects metal speciation as well as other

salts in solution that may determine bioavailability of the 

metal. Statistical models such as REDEQL-EPAK can be 

used to estimate the amounts of various species of metal 

in a particular test water, thus improving the interpretation of toxicity data (Palawski et al. 1985).

Acute toxicity tests can be used for a variety of purposes, one of which is the estimation of the half-life of 

biological activity of a toxicant (Marking 1972; Marking 

and Dawson 1972). Two approaches were outlined, one 

calling for the use of only one species of fish and aged 

solutions and the other involving aged solutions and 

several species of fish, each with different sensitivities 

to the toxic chemical (Marking 1972). The resulting LC50 

data can be used to estimate the half-life of biological activity or to calculate a deactivation index. A deactivation 

index greater than 1.0 indicates loss of biological activity, and an index of less than 1.0 indicates an increase 

in toxicity of the aged solution over time.

Another application of acute toxicity tests is in on-site 

toxicity assessment. For example, on-site tests have been 

used in sea lamprey control in the Great Lakes basin 

(Howell and Marquette 1962) to establish concentrations 

of fish toxicants for use in fish control (Burress 1975), 

to measure the toxicity of effluents (Norberg and Mount

1985), and to monitor water quality (Hall et al. 1985; 

Susan E. Finger, personal communication). Most tests involve static or static-renewal exposure systems but a few 

use a flow-through arrangement (Fig. 3).

The acute toxicity of sediments can be assessed by using 

a liquid phase elutriate test or solid phase tests in which 

fish are exposed directly to the sediment and water or to 

water flowing over the sediment, in a modified static test 

system based on the technique of Fremling (1975). The 

elutriate or liquid phase testing has shown little promise 

in defining the toxicity of sediments (Dillon and Gibson

1986). Marking et al. (1984) tested suspended sediments 

against fish to define the potential toxicity of sediments 

suspended by commercial boat traffic on the Upper Mississippi River system. Prater and Anderson (1977) used 

the modified Fremling (1975) assay system to assess the 

96-h toxicity of harbor sediments. A further modification 

of this system was discussed by LeBlanc and Surprenant 

(1985) in an assessment of toxicity of contaminated 

freshwater sediments. Sediment toxicity testing remains 

a subject of active research on the development of methods 

and the standardization of test protocols.

In the last 15 years, interest has grown in assessing the 

toxicity of mixtures of chemicals to fish. Marking and 

Dawson (1975) published a method of assessing the toxicity of mixtures of chemicals by using an additivity index in which additive values were represented by zero,
Fig. 3. Chemist evaluating the toxicity of river water to larval striped bass using an on-site flow-through toxicity test system. 

Photo by P. Winger.

greater than additive values by positive numbers, and less 

than additive values by negative numbers. The method 

for assessing additive toxicity of chemical mixtures was 

further defined by Marking (1977, 1985). Konemann 

(1981) proposed the use of a mixture toxicity index. This 

scale for measuring the toxicity of mixtures fulfills two 

requisites he outlined: (1) the scale should give, independently from the number of compounds in the mixture and the ratio between the concentrations, constant 

values for two reference points no addition (zero), and 

concentration addition (1.0); and (2) it should have a 

logarithmic form because of the log-normal distribution

of LCSO's. Konemann (1981) proposed the use of the mixture toxicity index for mixtures of two or more chemicals. 

Broderius and Kahl (1985) demonstrated concentration 

additive, acute joint toxicity for 27 organic chemicals from 

7 different classes, using mixtures of as many as 

21 chemicals. Their results are consistent with those of 

other investigators and suggest that numerous industrial 

chemicals are likely to kill fish through a similar narcosis 

type of toxic action.

Also during the last 15 years, data generated from acute 

toxicity tests have been increasingly used to develop quantitative structure-activity relations (QSAR's) in an effort
to predict the toxicity of pollutants (chemicals) from their Drug Administration and the Environmental Protection

physiochemical properties (Lipnick et al. 1985; Hermens Agency (Meyer and Schnick 1980). The conduct of some

1986; Kaiser 1987). Hermens (1986) suggested that acute aquatic toxicity tests now falls under the Good

QSAR's will eventually replace range-finding tests and Laboratory Practice Standards (GLPS; Federal Register

will be used for several other purposes: to set priorities 1987). The GLPS also apply to studies conducted for the

for testing existing compounds, to predict the aquatic tox- Federal Insecticide, Fungicide and Rodenticide Act and

icity of pesticide products during their development, to Toxic Substances Control Act. In addition, the use of acute

classify large numbers of compounds into a limited num- toxicity data plays an integral role in ecological risk assessber of groups, and to estimate the joint toxicity of mixtures, ment procedures (Urban and Cook 1986). Static acute toxData from acute tests are also used to register the uses icity tests will thus continue to play an essential but limited

of various chemicals for fisheries work by the Food and role in aquatic toxicology in the future.

Table. Fish toxicity testing (1863-1987): a brief review (year, investigators, significant event, and reference).

1863

C. Penny and C. Adams

First recorded toxicity tests with fish. More than 400 tests with 71 chemicals in river water and test solutions. (Jones 

1964) 

1917

E. B. Powers

Recommended the goldfish as a test animal and introduced the concept of measuring survival time versus concentration. (Powers 1917) 

1919-30

K. E. Carpenter

Used caged fishes to document toxicity of runoff from lead and zinc mine tailings. In studies with 13 species of 

freshwater fishes, showed that exposure of fish to lead salts in distilled water resulted in similar symptoms of intoxication. (Carpenter 1930)

1927

D. L. fielding

First published critical analyses of conditions necessary for successful testing of polluting substances against fish, 

(fielding 1927)

1928

P. Steinmann

First recordings of respiration and heart rate of trout exposed to toxic concentrations of chemicals. (Steinmann 1928)

1937

M. M. Ellis

Classic paper on "Detection and measurement of stream pollution" brought together most of knowledge on mode 

of action of toxicants. Also includes extensive laboratory toxicity tests with goldfish and Daphnia magna. (Ellis 1937)

1938

J. R. E. Jones

Documented the negative effect of calcium on the toxicity of lead and zinc to the stickleback (Gasterosteus aculeatus)

in soft water. (Jones 1938) 

1942

Introduction of DDT as an "ideal" insecticide. (Dunlap 1981) 

1948

C. M. Tarzwell

U.S. Public Health Service established Biology Section in Cincinnati, Ohio, laboratory to study water pollution 

problems. (C. M. Tarzwell, personal communication, 1985) 

1950

P. Doudoroff and M. Katz

Critical review of literature on the toxicity of industrial wastes and their components to fish. Parts I and n (Doudoroff 

and Katz 1950, 1953)
Table. Continued.

1951

P. Doudoroff and committee members

Development of standardized criteria for acute toxicity tests began in earnest and culminated in the 1970's with 

the acceptance of standardized testing criteria. (Doudoroff et al. 1951; P. Doudoroff, personal communication, 1986) 

1952-54

E. M. Wood, E. Hollis, and R. E. Lennon

Publication of the largest data set on the toxicity of chemicals to freshwater fish to that time. More than 4,000 chemicals 

were screened against six species of fish. (Wood 1953; Hollis and Lennon 1954)

1952

A. T. James and A. J. P. Martin

Established gas-liquid partition chromatography as an analytical technique. (Ettre 1975)

1953

V. C. Applegate and coworkers

The screening of chemicals for use in the control of the sea lamprey in the Great Lakes drainage began in 1953 

at the Hammond Bay Biological Station. More than 4,000 chemicals were tested before the discovery of the selective lamprey larvicide, 3-trifluoromethyl-4-nitrophenol. (Applegate et al. 1957; Howell et al. 1980) 

1959

R. E. Lennon

Opening of the Fish Control Laboratory in LaCrosse, Wisconsin, to investigate physical, chemical and biological 

methods for the control of nuisance fishes. (R. E. Lennon, personal communication, 1985)

1959

O. B. Cope

Establishment of the Fish-Pesticide Research Laboratory at Denver, Colorado. (O. B. Cope, personal communication, 1985)

1960

C. M. Tarzwell and co workers

The capacity of the Biology Section of the U.S. Public Health Service. Laboratory to do toxicity testing with fish 

greatly expanded at the Newtown (Ohio) State Fish Hatchery. (Tarzwell 1978) 

1960

Standard Bioassay Methods Committee

Publication of "Bioassay methods for the evaluation of acute toxicity of industrial wastes and other substances to 

fish" in the llth edition of Standard Methods. (American Public Health Association et al. 1960) 

1975

Eleven-member committee

Publication of Methods for Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians. (U.S. Environmental 

Protection Agency 1975) 

1980

Subcommittee E 35.23 on Safety of Aquatic Organisms

Approval and publication of Standard Practice for Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates, 

and Amphibians. (American Society for Testing and Materials 1980)

Acknowledgments American Public Health Association, American Water Works

Association, and Water Pollution Control Federation. 1960.

I thank P. Doudoroff, R. E. Lennon, and C M Tarz- Standard methods for the examination of water and wastewell for contributing to this review. ¥*? "* ^ American ^^ Health Association, New 

& York. 626 pp.

American Public Health Association, American Water Works

P r- Association, and Water Pollution Control Federation. 1981. 

K.e Terences Standard methods for examination of water and wastewater.

15th ed. American Public Health Association, Washington, 

Alien, J. L., and J. B. Hunn. 1986. Fate and distribution studies D.C. 1134 pp.

of some drugs used in aquaculture. Vet. Hum. Toxicol. American Society for Testing and Materials. 1980. Standard 

28(Supp.) 1:21 -24. practice for conducting acute toxicity tests with fishes, macro-
invertebrates, and amphibians. E729-80. American Society 

for Testing and Materials, Philadelphia, Pa. 25 pp.

Applegate, V. C., J. H. Howell, A. E. Hall, Jr., and M. A. 

Smith. 1957. Toxicity of 4,346 chemicals to larval lampreys 

and fishes. U.S. Fish Wildl. Serv., Spec. Sci. Rep. Fish. 

207. 157 pp.

Belding, D. L. 1927. Toxicity experiments with fish in reference 

to trade waste pollution. Trans. Am. Fish. Soc. 57:100-119.

Broderius, S., and M. Kahl. 1985. Acute toxicity of organic 

chemical mixtures to the fathead minnow. Aquat. Toxicol. 

6:307-322.

Brown, V. M. 1973. Concepts and outlook in testing the toxicity of substances to fish. Pages 73-95 in G. E. Glass, ed. 

Bioassay techniques and environmental chemistry. Ann Arbor 

Science Publishers Inc., Ann Arbor, Mich.

Burress, R. M. 1975. Development and evaluation of on-site 

toxicity test procedures for fishery investigations. U.S. Fish 

Wildl. Serv., Invest. Fish Control 68. 8 pp.

Carpenter, K. E. 1925. On the biological factor involved in the 

destruction of river-fisheries by pollution due to lead mining. 

Ann. Appl. Biol. 12:1-13.

Carpenter, K. E. 1930. Further reaches on the action of metallic 

salts on fishes. J. Exp. Zool. 56:407-422.

Committee on Methods for Toxicity Tests with Aquatic 

Organisms. 1975. Methods for acute toxicity tests with fish, 

macroinvertebrates, and amphibians. U.S. Environmental 

Protection Agency, Ecological Research Series, EPA660/3-75-009. 61 pp.

Dillon, T. M., and A. B. Gibson. 1986. Bioassessment 

methodologies for the regulatory testing of freshwater dredged 

material; proceedings of a workshop. Miscellaneous paper 

EL-86-6. U.S. Army Engineers Waterways Experiment Station, Vicksburg, Miss.

Doudoroff, P., and M. Katz. 1950. Critical review of literature 

on the toxicity of industrial wastes and their components to 

fish. I. Alkalies, acids, and inorganic gasses. Sewage Ind. 

Wastes 22:1432-1458.

Doudoroff, P., and M. Katz. 1953. Critical review of literature 

on the toxicity of industrial wastes and their components to 

fish. H. The metals, as salts. Sewage Ind. Wastes 25:802-839.

Doudoroff, P., B. G. Anderson, G. E. Burdick, P. S. Galtsoff, 

W. B. Hart, R. Patrick, E. R. Strong, E. W. Surber, and 

W. M. Van Horn. 1951. Bioassay methods for the evaluation of acute toxicity of industrial wastes to fish. Sewage Ind. 

Wastes 23:1380-1397.

Dunlap, T. R. 1981. DOT scientists, citizens, and public policy. 

Princeton University Press, Princeton, N.J. 318 pp.

Ellis, M. M. 1937. Detection and measurement of stream pollution. Bull. U.S. Bur. Fish. 48:365-437.

Ettre, L. S. 1975. The development of gas chromatography. 

J. Chromatogr. 112:1-26.

Federal Register. 1987. Good laboratory practice standards. 40 

CFR Parts 160 and 792, Vol. 52, No. 248:48920-48946.

Fremling, C. R. 1975. Acute toxicity of the lampricide 

3-trifluoromethyl-4-nitrophenol (TFM) to nymphs of mayflies 

(Hexagenia sp.). U.S. Fish Wildl. Serv., Invest. Fish Control 58. 8 pp.

Gelber, R. D., T. Lavin, C. R. Mehta, and D. A. Schoenfeld. 

1985. Statistical analysis. Pages 110-123 in G. M. Rand and 

S. R. Petrocelli, eds. Fundamentals of aquatic toxicology. 

Hemisphere Publishing Corp., New York.

Hall, L. W., Jr., A. E. Pinkney, L. O. Horseman, and S. E. 

Finger. 1985. Mortality of striped bass larvae in relation to 

contaminants and water quality in a Chesapeake Bay tributary. 

Trans. Am. Fish. Soc. 114:861-868.

Hart, W. B., P. Doudoroff, and J. Greenbank. 1945. The evaluation of the toxicity of industrial wastes, chemicals and other 

substances to freshwater fishes. Waste Control Laboratory, 

The Atlantic Refining Company, Philadelphia, Pa. 332 pp.

Hermens, J. L. M. 1986. Quantitative structure-activity relationships in aquatic toxicology. Pestic. Sci. 17:287-296.

Hollis, E. H., and R. E. Lennon. 1954. The toxicity of 1,085 

chemicals to fish. Fish Toxicity Project Report No. 2. U.S. 

Fish and Wildlife Service, Kearneysville, W. Va. 66 pp. 

(Reprinted 1981, U.S. Environ. Protect. Agency, Off. Toxic Subst., EPA 560/6-87-002, Part 2.)

Howell, J. H., J. J. Lech, and J. L. Alien. 1980. Development 

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560/6-87-002, Part 1.)
Hunn, Joseph B. 1989. History of Acute Toxicity Tests with Fish, 1863-1987.

U.S. Fish Wildl. Serv., Invest. Fish Control 98. 10 pp.

Acute toxicity tests with fish were first reported in 1863. Test methodology 

developed slowly before World War II but after the war, considerable effort 

was made to standardize test methods. A standardized method was published 

in 1960, refined in 1975, and updated in 1980. The primary use of the static 

acute toxicity test is to determine the median concentrations of chemicals that 

are lethal to aquatic organisms. Static acute toxicity tests will continue to play 

an essential role in aquatic toxicology in the foreseeable future.

Key words: Acute toxicity, static tests, fish, history.

Hunn, Joseph B. 1989. History of Acute Toxici

U.S. Fish Wildl. Serv., Invest. Fish Contro

Acute toxicity tests with fish were first repo

developed slowly before World War U but af

was made to standardize test methods. A stan

in 1960, refined in 1975, and updated in 1980

acute toxicity test is to determine the median c

are lethal to aquatic organisms. Static acute tox

an essential role in aquatic toxicology in the f

Key words: Acute toxicity, static tests, fish, 

Hunn, Joseph B. 1989. History of Acute Toxicity Tests with Fish, 1863-1987. U.S. Fish Wildl. 

Serv., Invest. Fish Control 98. 10 pp.

Acute toxicity tests with fish were first reported in 1863. Test methodology developed slowly before 

World War II but after the war, considerable effort was made to standardize test methods. A standardized method was published in 1960, refined in 1975, and updated in 1980. The primary use of the 

static acute toxicity test is to determine the median concentrations of chemicals that are lethal to aquatic 

organisms. Static acute toxicity tests will continue to play an essential role in aquatic toxicology in 

the foreseeable future.

Key words: Acute toxicity, static tests, fish, history.

Hunn, Joseph B. 1989. History of Acute Toxicity Tests w

Serv., Invest. Fish Control 98. 10 pp.

Acute toxicity tests with fish were first reported in 1863. Te

World War II but after the war, considerable effort was made 

ized method was published in 1960, refined in 1975, and up

static acute toxicity test is to determine the median concentratio

organisms. Static acute toxicity tests will continue to play a

the foreseeable future.

Key words: Acute toxicity, static tests, fish, history.

.'.- U S Government Printing Office: 1989 673 208/00008



(Reports 87 through 89 are in one cover.)

87. Ethyl-p-aminobenzoate (Benzocaine): Efficacy as an Anesthetic for Five Species of Freshwater Fish, by V. K. Dawson 

and P. A. Gilderhus. 1979. 5 pp.

88. Influences of Selected Environmental Factors on the Activity of a Prospective Fish Toxicant, 2-(Digeranyl-amino)-ethanol, 

in Laboratory Tests, by C. A. Launer and T. D. Bills. 1979. 4 pp.

89. Toxicities of the Lampricides 3-Trifluoromethyl-4-nitrophenol (TFM) and the 2-Aminoethanol Salt of 2',5-Dichloro-4'- 

nitrosalicylanilide (Bayer 73) to Four Bird Species, by R. H. Hudson. 1979. 5 pp.

(Reports 90 and 91 are in one cover.)

90. Accumulation and Loss of 2',5-Dichloro-4'-nitrosalicylanilide (Bayer 73) by Fish: Laboratory Studies, by V. K. Dawson, 

J. B. Sills, and Charles W. Luhning. 1982. 5 pp.

91. Effects of Synergized Rotenone on Nontarget Organisms in Ponds, by R. M. Burress. 1982. 7 pp.

(Reports 92 through 94 are in one cover.)

92. Acute and Chronic Toxicity of Rotenone to Daphnia magna, by J. J. Rach, T. D. Bills, and L. L. Marking. 1988. 5 pp.

93. Toxicity of Rotenone to Developing Rainbow Trout, by T. D. Bills, J. J. Rach, and L. L. Marking. 1988. 3 pp.

94. Oral Toxicity of Rotenone to Mammals, by L. L. Marking. 1988. 5 pp.

95. Deposition and Persistence of Rotenone in Shallow Ponds During Cold and Warm Seasons, by P. A. Gilderhus, V. K. 

Dawson, and J. L. Alien. 1988. 7 pp.

(Reports 96 and 97 are in one cover.)

96. Effects of Environmental Factors on the Toxicity of Chloramine-T to Fish, by T. D. Bills, L. L. Marking, V. K. Dawson, 

and J. J. Rach. 1988. 6 pp.

97. Effects of Organic Matter and Loading Rates of Fish on the Toxicity of Chloramine-T, by T. D. Bills, L. L. Marking, 

V. K. Dawson, and G. E. Howe. 1988. 4 pp.

NOTE: Use of trade names does not imply U.S. Government endorsement of commercial products.
TAKE PRIDE

in America

US. DEPARTMENT OF THE INTERIOR

FISH AND WILDLIFE SERVICE

us. ^

FISH * WILDLIFE 

SERVICE

As the Nation's principal conservation agency, the Department of the Interior has responsibility for 

most of our nationally owned public lands and natural resources. This includes fostering the wisest 

use of our land and water resources, protecting our fish and wildlife, preserving the environmental 

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life through outdoor recreation. The Department assesses our energy and mineral resources and works 

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a major responsibility for American Indian reservation communities and for people who live in island 

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