ASTM E729-23e1
(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...
General Information
Standards Content (Sample)
This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
´1
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
ε NOTE—Sections 3 and 4 were editorially reorganized, and 7.8 and 8.2.1.1 were editorially corrected in February 2023.
1. Scope materials that have a high oxygen demand, are highly volatile,
2 are rapidly transformed biologically or chemically in aqueous
1.1 This guide (1) describes procedures for obtaining
solution, or are removed from test solutions in substantial
laboratory data concerning the adverse effects (for example,
quantities by the test chambers or organisms during the test.
lethality and immobility) of a test material added to dilution
Because the pH and concentrations of dissolved oxygen and
water, but not to food, on certain species of freshwater and
test material are maintained at desired levels and degradation
saltwater fishes, macroinvertebrates, and amphibians, usually
and metabolic products are removed, tests using renewal and
during 2 to 4-day exposures, depending on the species. These
flow-through methods are preferable; test organisms may be
procedures will probably be useful for conducting acute
fed during renewal and flow-through tests. Although renewal
toxicity tests with many other aquatic species, although modi-
tests might be more cost-effective, flow-through tests are
fications might be necessary.
generally preferable.
1.2 Other modifications of these procedures might be justi-
1.4 Acute tests may be performed to meet regulatory data
fied by special needs or circumstances such as meeting specific
requirements or to obtain time-independent estimates of toxic-
study goals, regulatory needs, or to accommodate specific test
ity.
organism life stages. Although using appropriate procedures is
1.4.1 If the objective is to obtain data to meet regulatory
more important than following prescribed procedures, results
of tests conducted using unusual or novel procedures are not requirements, it may be necessary to limit the number of
observation times based on stipulations of the regulatory
likely to be comparable to results of many other tests. Com-
parison of results obtained using modified and unmodified agency and cost considerations.
versions of these procedures might provide useful information
1.4.2 If the objective of an acute toxicity test is to determine
concerning new concepts and procedures for conducting acute
a time-independent (that is, incipient, threshold, or asymptotic)
tests.
toxicity level, an appropriate number of observations must be
taken over an exposure duration of sufficient length to establish
1.3 This guide describes tests using three basic exposure
the shape of the toxicity curve or allow the direct or math-
techniques: static, renewal, and flow-through. Selection of the
ematically estimated determination of a time-independent tox-
technique to use in a specific situation will depend on the needs
icity value (1), or both.
of the investigator and on available resources. Tests using the
static technique provide the most easily obtained measure of
1.5 In the development of these procedures, an attempt was
acute toxicity, but conditions often change substantially during
made to balance scientific and practical considerations and to
static tests; therefore, static tests should not last longer than 96
ensure that the results will be sufficiently accurate and precise
h, and test organisms should not be fed during such tests unless
for the applications for which they are commonly used. A
the test organisms are severely stressed without feeding over
major consideration was that the common uses of the results of
48 h. Static tests should probably not be conducted on
acute toxicity tests do not require or justify stricter require-
ments than those set forth herein. Although the tests may be
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental improved by using more organisms, longer acclimation times,
Assessment, Risk Management and Corrective Actionand is the direct responsibility
and so forth, the requirements presented herein should usually
of Subcommittee E50.47 on Biological Effects and Environmental Fate.
be sufficient.
Current edition approved Jan. 1, 2023. Pu
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