ASTM E729-23
(Guide)Standard Guide for Conducting Acute Toxicity Tests on Test Materials with Fishes, Macroinvertebrates, and Amphibians
Standard Guide for Conducting Acute Toxicity Tests on Test Materials with Fishes, Macroinvertebrates, and Amphibians
SIGNIFICANCE AND USE
5.1 An acute toxicity test is conducted to obtain information concerning the immediate effects on test organisms of a short-term exposure to a test material under specific experimental conditions. An acute toxicity test does not provide information about whether delayed effects will occur, although a post-exposure observation period, with appropriate feeding, if necessary, might provide such information. Bioavailability of the test substance may also differ between real-world exposures and laboratory exposures due to site-specific water quality conditions (see Guides E1192, E1563, and E2455).
5.2 Results of acute toxicity tests might be used to predict acute effects likely to occur on aquatic organisms in field situations as a result of exposure under comparable conditions, except that (1) motile organisms might avoid exposure when possible, and (2) toxicity to benthic organisms might be dependent on sorption or settling of the test material onto the substrate.
5.3 Results of acute tests might be used to compare the acute sensitivities of different species and the acute toxicities of different test materials, and to study the effects of various environmental factors on results of such tests.
5.4 Results of acute toxicity tests might be an important consideration when assessing the hazards of materials to aquatic organisms (see Guide E1023) or when deriving water quality criteria for aquatic organisms (3).
5.5 Results of acute toxicity tests might be useful for studying the biological availability of, and structure-activity relationships between, test materials.
5.6 Results of acute toxicity tests will depend on the temperature, composition of the dilution water, condition of the test organisms, exposure technique, and other factors.
SCOPE
1.1 This guide (1)2 describes procedures for obtaining laboratory data concerning the adverse effects (for example, lethality and immobility) of a test material added to dilution water, but not to food, on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, usually during 2 to 4-day exposures, depending on the species. These procedures will probably be useful for conducting acute toxicity tests with many other aquatic species, although modifications might be necessary.
1.2 Other modifications of these procedures might be justified by special needs or circumstances such as meeting specific study goals, regulatory needs, or to accommodate specific test organism life stages. Although using appropriate procedures is more important than following prescribed procedures, results of tests conducted using unusual or novel procedures are not likely to be comparable to results of many other tests. Comparison of results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting acute tests.
1.3 This guide describes tests using three basic exposure techniques: static, renewal, and flow-through. Selection of the technique to use in a specific situation will depend on the needs of the investigator and on available resources. Tests using the static technique provide the most easily obtained measure of acute toxicity, but conditions often change substantially during static tests; therefore, static tests should not last longer than 96 h, and test organisms should not be fed during such tests unless the test organisms are severely stressed without feeding over 48 h. Static tests should probably not be conducted on materials that have a high oxygen demand, are highly volatile, are rapidly transformed biologically or chemically in aqueous solution, or are removed from test solutions in substantial quantities by the test chambers or organisms during the test. Because the pH and concentrations of dissolved oxygen and test material are maintained at desired levels and degradation and metabolic products are removed, tests using ren...
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Designation: E729 − 23
Standard Guide for
Conducting Acute Toxicity Tests on Test Materials with
1
Fishes, Macroinvertebrates, and Amphibians
This standard is issued under the fixed designation E729; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
This standard has been approved for use by agencies of the U.S. Department of Defense.
1. Scope are rapidly transformed biologically or chemically in aqueous
2 solution, or are removed from test solutions in substantial
1.1 This guide (1) describes procedures for obtaining
quantities by the test chambers or organisms during the test.
laboratory data concerning the adverse effects (for example,
Because the pH and concentrations of dissolved oxygen and
lethality and immobility) of a test material added to dilution
test material are maintained at desired levels and degradation
water, but not to food, on certain species of freshwater and
and metabolic products are removed, tests using renewal and
saltwater fishes, macroinvertebrates, and amphibians, usually
flow-through methods are preferable; test organisms may be
during 2 to 4-day exposures, depending on the species. These
fed during renewal and flow-through tests. Although renewal
procedures will probably be useful for conducting acute
tests might be more cost-effective, flow-through tests are
toxicity tests with many other aquatic species, although modi-
generally preferable.
fications might be necessary.
1.4 Acute tests may be performed to meet regulatory data
1.2 Other modifications of these procedures might be justi-
requirements or to obtain time-independent estimates of toxic-
fied by special needs or circumstances such as meeting specific
ity.
study goals, regulatory needs, or to accommodate specific test
1.4.1 If the objective is to obtain data to meet regulatory
organism life stages. Although using appropriate procedures is
requirements, it may be necessary to limit the number of
more important than following prescribed procedures, results
observation times based on stipulations of the regulatory
of tests conducted using unusual or novel procedures are not
agency and cost considerations.
likely to be comparable to results of many other tests. Com-
1.4.2 If the objective of an acute toxicity test is to determine
parison of results obtained using modified and unmodified
versions of these procedures might provide useful information a time-independent (that is, incipient, threshold, or asymptotic)
concerning new concepts and procedures for conducting acute toxicity level, an appropriate number of observations must be
tests. taken over an exposure duration of sufficient length to establish
the shape of the toxicity curve or allow the direct or math-
1.3 This guide describes tests using three basic exposure
ematically estimated determination of a time-independent tox-
techniques: static, renewal, and flow-through. Selection of the
icity value (1), or both.
technique to use in a specific situation will depend on the needs
of the investigator and on available resources. Tests using the
1.5 In the development of these procedures, an attempt was
static technique provide the most easily obtained measure of
made to balance scientific and practical considerations and to
acute toxicity, but conditions often change substantially during
ensure that the results will be sufficiently accurate and precise
static tests; therefore, static tests should not last longer than 96
for the applications for which they are commonly used. A
h, and test organisms should not be fed during such tests unless
major consideration was that the common uses of the results of
the test organisms are severely stressed without feeding over
acute toxicity tests do not require or justify stricter require-
48 h. Static tests should probably not be conducted on
ments than those set forth herein. Although the tests may be
materials that have a high oxygen demand, are highly volatile,
improved by using more organisms, longer acclimation times,
and so forth, the requirements presented herein should usually
be sufficient.
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental
1.6 Results of acute toxicity tests should usually be reported
Assessment, Risk Management and Corrective Actionand is the direct responsibility
in terms of an LC50 (median lethal concentration) or EC50
of Subcommittee E50.47 on Biological Effects and Environmental Fate.
Current edition approved Jan. 1, 2023. Published February 2023. Originally
(median effective concentration) at the end of the test, but it is
approved in 1980. Last previous edition appr
...
This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: E729 − 96 (Reapproved 2014) E729 − 23
Standard Guide for
Conducting Acute Toxicity Tests on Test Materials with
1
Fishes, Macroinvertebrates, and Amphibians
This standard is issued under the fixed designation E729; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
This standard has been approved for use by agencies of the U.S. Department of Defense.
1. Scope
2
1.1 This guide (1) describes procedures for obtaining laboratory data concerning the adverse effects (for example, lethality and
immobility) of a test material added to dilution water, but not to food, on certain species of freshwater and saltwater fishes,
macroinvertebrates, and amphibians amphibians, usually during 2 to 8-day4-day exposures, depending on the species. These
procedures will probably be useful for conducting acute toxicity tests with many other aquatic species, although modifications
might be necessary.
1.2 Other modifications of these procedures might be justified by special needs or circumstances. circumstances such as meeting
specific study goals, regulatory needs, or to accommodate specific test organism life stages. Although using appropriate procedures
is more important than following prescribed procedures, results of tests conducted using unusual or novel procedures are not likely
to be comparable to results of many other tests. Comparison of results obtained using modified and unmodified versions of these
procedures might provide useful information concerning new concepts and procedures for conducting acute tests.
1.3 This guide describes tests using three basic exposure techniques: static, renewal, and flow-through. Selection of the technique
to use in a specific situation will depend on the needs of the investigator and on available resources. Tests using the static technique
provide the most easily obtained measure of acute toxicity, but conditions often change substantially during static tests; therefore,
static tests should not last longer than 96 h, and test organisms should not be fed during such tests. tests unless the test organisms
are severely stressed without feeding over 48 h. Static tests should probably not be conducted on materials that have a high oxygen
demand, are highly volatile, are rapidly transformed biologically or chemically in aqueous solution, or are removed from test
solutions in substantial quantities by the test chambers or organisms during the test. Because the pH and concentrations of
dissolved oxygen and test material are maintained at desired levels and degradation and metabolic products are removed, tests
using renewal and flow-through methods are preferable and may last longer than 96 h; preferable; test organisms may be fed during
renewal and flow-through tests. Although renewal tests might be more cost-effective, flow-through tests are generally preferable.
1.4 Acute tests may be performed to meet regulatory data requirements or to obtain time-independent estimates of toxicity.
1.4.1 If the objective is to obtain data to meet regulatory requirements, it may be necessary to limit the number of observation
times based on stipulations of the regulatory agency and cost considerations.
1.4.2 If the objective of an acute toxicity test is to determine a time-independent (that is, incipient, threshold, or asymptotic)
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental Assessment, Risk Management and Corrective Actionand is the direct responsibility of
Subcommittee E50.47 on Biological Effects and Environmental Fate.
Current edition approved Oct. 1, 2014Jan. 1, 2023. Published December 2014February 2023. Originally approved in 1980. Last previous edition approved in 20072014
as E729 – 96(2007). DOI: 10.1520/E0729-96R14.(2014). DOI: 10.1520/E0729-23.
2
The boldface numbers in parentheses refer to the list of references at the end of this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
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E729 − 23
toxicity level, an appropriate number of observations must be taken over an exposure duration of sufficient length to establish the
shape of the toxicity curve or allow the direct or mathematically estimated determination of a time-independent toxicity value (1),
or bo
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