ASTM E2591-22
(Guide)Standard Guide for Conducting Whole Sediment Toxicity Tests with Amphibians
Standard Guide for Conducting Whole Sediment Toxicity Tests with Amphibians
SIGNIFICANCE AND USE
5.1 While federal criteria and state standards exist that define acute and chronic “safe” levels in the water column, effects levels in the sediment are poorly defined and may be dependent upon numerous modifying factors. Even where USEPA recommended Water Quality Criteria (WQC, (49)) are not exceeded by water-borne concentrations, organisms that live in or near the sediment may still be adversely affected (50). Therefore, simply measuring the concentration of a chemical in the sediment or in the water is often insufficient to evaluate its actual environmental toxicity. Concentrations of contaminants in sediment may be much higher than concentrations in overlying water; this is especially true of hydrophobic organic compounds as well as inorganic ions that have a strong affinity for organic ligands and negatively-charged surfaces. Higher chemical concentrations in sediment do not, however, always translate to greater toxicity or bioaccumulation (51), although research also suggests that amending sediment with organic matter actually increases the bioaccumulation of contaminant particles (52, 53). Other factors that can potentially influence sediment bioaccumulation and toxicity include pH mineralogical composition, acid-volatile sulfide (AVS) grain size, and temperature (54-56). Laboratory toxicity tests provide a direct and effective way to evaluate the impacts of sediment contamination on environmental receptors while providing empirical consideration of all of the physical, chemical and biological parameters that may influence toxicity.
5.2 Amphibians are often a major ecosystem component of wetlands around the world, however limited data are available regarding the effects of sediment-bound contaminants to amphibians (39, 41, 43, 55, 57, 58). Laboratory studies such as the procedure described in this standard are one means of directly assessing sediment toxicity to amphibians in order to evaluate potential ecological risks in wetlands.
5.3 Results from sed...
SCOPE
1.1 This standard covers procedures for obtaining laboratory data concerning the toxicity of test material (for example, sediment or hydric soil (that is, a soil that is saturated, flooded, or ponded long enough during the growing season to develop anaerobic (oxygen-lacking) conditions that favor the growth and regeneration of hydrophytic vegetation)) to amphibians. This test procedure uses larvae of the northern leopard frog (Lithobates pipiens). Other anuran species (for example, the green frog (Lithobates clamitans), the wood frog (Lithobates sylvatica), the American toad (Bufo americanus)) may be used if sufficient data on handling, feeding, and sensitivity are available. Test material may be sediments or hydric soil collected from the field or spiked with compounds in the laboratory.
1.2 The test procedure describes a 10-d whole sediment toxicity test with an assessment of mortality and selected sublethal endpoints (that is, body width, body length). The toxicity tests are conducted in 300 to 500-mL chambers containing 100 mL of sediment and 175 mL of overlying water. Overlying water is renewed daily and larval amphibians are fed during the toxicity test once they reach Gosner stage 25 (operculum closure over gills). The test procedure is designed to assess freshwater sediments, however, R. pipiens can tolerate mildly saline water (not exceeding about 2500 mg Cl-/L, equivalent to a salinity of about 4.1 when Na+ is the cation) in 10-d tests, although such tests should always include a concurrent freshwater control. Alternative test durations and sublethal endpoints may be considered based on site-specific needs. Statistical evaluations are conducted to determine whether test materials are significantly more toxic than the laboratory control sediment or a field-collected reference sample(s).
1.3 Where appropriate, this standard has been designed to be consistent with previously developed methods for assessing s...
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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.
Designation: E2591 − 22
Standard Guide for
Conducting Whole Sediment Toxicity Tests with
1
Amphibians
This standard is issued under the fixed designation E2591; 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.
1. Scope sediment toxicity to invertebrates (for example, Hyalella az-
teca and Chironomus dilutus toxicity tests) described in the
1.1 This standard covers procedures for obtaining labora-
2
UnitedStatesEnvironmentalProtectionAgency(USEPA, (1))
tory data concerning the toxicity of test material (for example,
freshwatersedimenttestingguidance,TestMethodsE1367and
sediment or hydric soil (that is, a soil that is saturated, flooded,
E1706, and Guides E1391, E1525, E1611, and E1688. Tests
or ponded long enough during the growing season to develop
extending to 10 d or beyond, and including sublethal measure-
anaerobic (oxygen-lacking) conditions that favor the growth
ments such as growth, are considered more effective in
and regeneration of hydrophytic vegetation)) to amphibians.
identifying chronic toxicity and thus delineating areas of
This test procedure uses larvae of the northern leopard frog
moderate contamination (1-3).
(Lithobates pipiens). Other anuran species (for example, the
green frog (Lithobates clamitans), the wood frog (Lithobates
1.4 Many historical amphibian studies, both water and
sylvatica), theAmerican toad (Bufo americanus)) may be used
sediment exposure, have used tests of shorter duration (5 days
if sufficient data on handling, feeding, and sensitivity are
or less) (for example, 4-7) and, although both survival and
available. Test material may be sediments or hydric soil
sublethal endpoints were often assessed, there is substantive
collected from the field or spiked with compounds in the
evidence that tests of longer duration are likely to be more
laboratory.
sensitive to some contaminants (8-10). Research performed to
1.2 The test procedure describes a 10-d whole sediment develop and validate this test protocol included long-term
(through metamorphosis) investigations and other researchers
toxicity test with an assessment of mortality and selected
sublethal endpoints (that is, body width, body length). The have also conducted long-duration tests with anurans (7-20).
Interestingly, some studies with anurans have shown signifi-
toxicity tests are conducted in 300 to 500-mL chambers
containing100mLofsedimentand175mLofoverlyingwater. cantly reduced growth (for example, whole body mass, snout-
Overlyingwaterisreneweddailyandlarvalamphibiansarefed vent length) can be detected earlier in a longer-term test (for
during the toxicity test once they reach Gosner stage 25 example, at 14-20 d), but cannot be statistically distinguished
(operculum closure over gills). The test procedure is designed
in older organisms later in the test (11, 14). In the development
to assess freshwater sediments, however, R. pipiens can toler- of these procedures, an attempt was made to balance the needs
-
ate mildly saline water (not exceeding about 2500 mg Cl /L,
of a practical assessment with the importance of assessing
+
equivalent to a salinity of about 4.1 when Na is the cation) in longer-term effects so that the results will demonstrate the
10-d tests, although such tests should always include a con-
needed accuracy and precision. The most recent sediment
current freshwater control. Alternative test durations and sub- toxicity testing protocols for invertebrates have encompassed
lethal endpoints may be considered based on site-specific
longer duration studies which allow the measurement of
needs. Statistical evaluations are conducted to determine
reproductiveendpoints (1, 21).Suchtests,becauseofincreased
whether test materials are significantly more toxic than the
sensitivity of the sublethal endpoints, may also be helpful in
laboratory control sediment or a field-collected reference evaluatingtoxicity.Fulllife-cyclestudieswithanurans(includ-
sample(s).
ing reproduction) are usually not feasible from either a
technical or monetary standpoint. However, if site-specific
1.3 Where appropriate, this standard has been designed to
information indicates that the contaminants present are likely
be consistent with previously developed methods for assessing
to affect other endpoints (including teratogenicity), then the
duration of the toxicity test may be increased through meta-
morphosis or additional sublethal endpoints may be measured
1
Thisguideisunderthejur
...
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: E2591 − 07 (Reapproved 2013) E2591 − 22
Standard Guide for
Conducting Whole Sediment Toxicity Tests with
1
Amphibians
This standard is issued under the fixed designation E2591; 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.
1. Scope
1.1 This standard covers procedures for obtaining laboratory data concerning the toxicity of test material (for example, sediment
or hydric soil (that is, a soil that is saturated, flooded, or ponded long enough during the growing season to develop anaerobic
(oxygen-lacking) conditions that favor the growth and regeneration of hydrophytic vegetation)) to amphibians. This test procedure
uses larvae of the northern leopard frog (RanaLithobates pipiens). Other anuran species (for example, the green frog
(RanaLithobates clamitans), the wood frog (RanaLithobates sylvatica), the American toad (Bufo americanus)) may be used if
sufficient data on handling, feeding, and sensitivity are available. Test material may be sediments or hydric soil collected from the
field or spiked with compounds in the laboratory.
1.2 The test procedure describes a 10-d whole sediment toxicity test with an assessment of mortality and selected sublethal
endpoints (that is, body width, body length). The toxicity tests are conducted in 300 to 500-mL chambers containing 100 mL of
sediment and 175 mL of overlying water. Overlying water is renewed daily and larval amphibians are fed during the toxicity test
once they reach Gosner stage 25 (operculum closure over gills). The test procedure is designed to assess freshwater sediments,
-
however, R. pipiens can tolerate mildly saline water (not exceeding about 2500 mg Cl /L, equivalent to a salinity of about 4.1 when
+
Na is the cation) in 10-d tests, although such tests should always include a concurrent freshwater control. Alternative test durations
and sublethal endpoints may be considered based on site-specific needs. Statistical evaluations are conducted to determine whether
test materials are significantly more toxic than the laboratory control sediment or a field-collected reference sample(s).
1.3 Where appropriate, this standard has been designed to be consistent with previously developed methods for assessing sediment
toxicity to invertebrates (for example, Hyalella azteca and Chironomus dilutus toxicity tests) described in the United States
2
Environmental Protection Agency (USEPA, (1)) freshwater sediment testing guidance, Test Methods E1367 and E1706, and
Guides E1391, E1525, E1611, and E1688. Tests extending to 10 d or beyond, and including sublethal measurements such as
growth, are considered more effective in identifying chronic toxicity and thus delineating areas of moderate contamination (1-3).
1.4 Many historical amphibian studies, both water and sediment exposure, have used tests of shorter duration (5 days or less) (for
example, 4-7) and, although both survival and sublethal endpoints were often assessed, there is substantive evidence that tests of
longer duration are likely to be more sensitive to some contaminants (88-10, 9). Research performed to develop and validate this
test protocol included long-term (through metamorphosis) investigations and other researchers have also conducted long-duration
tests with anurans (7-1120). Interestingly, some studies with anurans have shown significantly reduced growth (for example, whole
body mass, snout-vent length) can be detected earlier in a longer-term test (for example, at 14-20 d), but cannot be statistically
distinguished in older organisms later in the test (11, 14). In the development of these procedures, an attempt was made to balance
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental Assessment, Risk Management and Corrective Action and is the direct responsibility
of Subcommittee E50.47 on Biological Effects and Environmental Fate.
Current edition approved March 1, 2013Jan. 1, 2022. Published March 2013April 2022. Originally approved in 2007. Last previous edition approved in 20072013 as
E2591E2591–07(2013).–07. DOI: 10.1520/E2591-07R13.10.1520/E2591-22.
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
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