ASTM E729-96(2014)
(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.
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 (2).
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 during 2 to 8-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. Although using appropriate procedures is more important than following prescribed procedures, results of tests conducted using unusual 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. 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; 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...
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Designation: E729 − 96 (Reapproved 2014)
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 concentrations of dissolved oxygen and test material are
2 maintained at desired levels and degradation and metabolic
1.1 This guide (1) describes procedures for obtaining
products are removed, tests using renewal and flow-through
laboratory data concerning the adverse effects (for example,
methods are preferable and may last longer than 96 h; test
lethality and immobility) of a test material added to dilution
organisms may be fed during renewal and flow-through tests.
water, but not to food, on certain species of freshwater and
Although renewal tests might be more cost-effective, flow-
saltwater fishes, macroinvertebrates, and amphibians during 2
through tests are generally preferable.
to 8-day exposures, depending on the species. These proce-
dures will probably be useful for conducting acute toxicity tests
1.4 Acute tests may be performed to meet regulatory data
with many other aquatic species, although modifications might
requirements or to obtain time-independent estimates of toxic-
be necessary.
ity.
1.4.1 If the objective is to obtain data to meet regulatory
1.2 Other modifications of these procedures might be justi-
requirements, it may be necessary to limit the number of
fied by special needs or circumstances. Although using appro-
observation times based on stipulations of the regulatory
priate procedures is more important than following prescribed
agency and cost considerations.
procedures, results of tests conducted using unusual procedures
are not likely to be comparable to results of many other tests. 1.4.2 If the objective of an acute toxicity test is to determine
a time-independent (that is, incipient, threshold, or asymptotic)
Comparison of results obtained using modified and unmodified
versions of these procedures might provide useful information toxicity level, an appropriate number of observations must be
taken over an exposure duration of sufficient length to establish
concerning new concepts and procedures for conducting acute
tests. the shape of the toxicity curve or allow the direct or math-
ematically estimated determination of a time-independent tox-
1.3 This guide describes tests using three basic exposure
icity value (1), or both.
techniques: static, renewal, and flow-through. Selection of the
technique to use in a specific situation will depend on the needs
1.5 In the development of these procedures, an attempt was
of the investigator and on available resources. Tests using the
made to balance scientific and practical considerations and to
static technique provide the most easily obtained measure of
ensure that the results will be sufficiently accurate and precise
acute toxicity, but conditions often change substantially during
for the applications for which they are commonly used. A
static tests; therefore, static tests should not last longer than 96
major consideration was that the common uses of the results of
h, and test organisms should not be fed during such tests. Static
acute toxicity tests do not require or justify stricter require-
tests should probably not be conducted on materials that have
ments than those set forth herein. Although the tests may be
a high oxygen demand, are highly volatile, are rapidly trans-
improved by using more organisms, longer acclimation times,
formed biologically or chemically in aqueous solution, or are
and so forth, the requirements presented herein should usually
removed from test solutions in substantial quantities by the test
be sufficient.
chambers or organisms during the test. Because the pH and
1.6 Results of acute toxicity tests should usually be reported
in terms of an LC50 (median lethal concentration) or EC50
(median effective concentration) at the end of the test, but it is
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental
desirable to provide information concerning the dependence of
Assessment, Risk Management and Corrective Actionand is the direct responsibility
adverse effects on both time and concentration. Thus, when
of Subcommittee E50.47 on Biological Effects and Environmental Fate.
Current edition approved Oct. 1, 2014. Published December 2014. Originally
feasible, flow-through and renewal tests should be conducted
approved in
...
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 2007) E729 − 96 (Reapproved 2014)
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 during 2 to 8-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. Although using appropriate
procedures is more important than following prescribed procedures, results of tests conducted using unusual 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. 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; 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)
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 both.
1.5 In the development of these procedures, an attempt was made to balance scientific and practical considerations and to ensure
that the results will be sufficiently accurate and precise for the applications for which they are commonly used. A major
consideration was that the common uses of the results of acute toxicity tests do not require or justify stricter requirements than
those set forth herein. Although the tests may be improved by using more organisms, longer acclimation times, and so forth, the
requirements presented herein should usually be sufficient.
1.6 Results of acute toxicity tests should usually be reported in terms of an LC50 (median lethal concentration) or EC50 (median
effective concentration) at the end of the test, but it is desirable to provide information concerning the dependence of adverse
effects on both time and concentration. Thus, when feasible, flow-through and renewal tests should be conducted so that LC50s
or EC50s can be reported f
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