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ASTM E2122-22

(Guide)

Standard Guide for Conducting In-situ Field Bioassays With Caged Bivalves

Standard Guide for Conducting In-situ Field Bioassays With Caged Bivalves

SIGNIFICANCE AND USE
5.1 The ecological importance of bivalves, their wide geographic distribution, ease of handling in the laboratory and the field, and their ability to filter and ingest large volumes of water and sediment particles make them appropriate species for conducting field bioassays to assess bioaccumulation potential and associated biological effects. The test procedures in this guide are intended to provide guidance for conducting controlled experiments with caged bivalves under “natural,” site-specific conditions. It is important to acknowledge that a number of “natural” factors can affect bivalve growth and the accumulation of chemicals in their tissues (Section 6, Interferences). This field bioassay can also be conducted in conjunction with laboratory bioassays to help answer questions raised in the field exposures. The field exposures can also be used to validate the results of laboratory bioassays.  
5.2 The ultimate resources of concern are communities. However, it is often difficult or impossible to adequately assess the ecological fitness or condition of the community or identify and test the most sensitive species. Bivalves are recommended as a surrogate test species for other species and communities for the following reasons: (1) They readily accumulate many chemicals and show sublethal effects associated with exposure to those chemicals (2); (2) they accumulate many chemicals through multiple pathways of exposure, including water, sediment, and food  (24, 25, 26, 27, 28, 29), and (3) caged bivalves have been shown to represent effects on the benthos more accurately than traditional laboratory tests (30, 31). Although bivalve species might be considered insensitive because of their wide use as indicators of chemical bioavailability, it has been suggested that sensitivity is related to the type of test, end points being measured, and duration of exposure  (2). In relatively short-term toxicity assessments in which survival is typically determined as the measureme...
SCOPE
1.1 This guide describes procedures for conducting controlled experiments with caged bivalves under field conditions. The purpose of this approach is to facilitate the simultaneous collection of field data to help characterize chemical exposure and associated biological effects in the same organism under environmentally realistic conditions. This approach of characterizing exposure and effects is consistent with the US EPA ecological risk assessment paradigm. Bivalves are useful test organisms for in-situ field bioassays because they (1) concentrate and integrate chemicals in their tissues and have a more limited ability to metabolize most chemicals than other species, (2) exhibit measurable sublethal effects associated with exposure to those chemicals, (3) provide paired tissue chemistry and response data which can be extrapolated to other species and trophic levels, (4) provide tissue chemistry data which can be used to estimate chemical exposure from water or sediment, and (5) facilitate controlled experimentation in the field with large sample sizes because they are easy to collect, cage, and measure (1, 2)2. The experimental control afforded by this approach can be used to place a large number of animals of a known size distribution in specific areas of concern to quantify exposure and effects over space and time within a clearly defined exposure period. Chemical exposure can be estimated by measuring the concentration of chemicals in water, sediment, or bivalve tissues, and effects can be estimated with survival, growth, and other sublethal end points (3). Although a number of assessments have been conducted using bivalves to characterize exposure by measuring tissue chemistry or associated biological effects, relatively few assessments have been conducted to characterize both exposure and biological effects simultaneously (2, 4, 5). This guide is specifically designed to help minimize the variability in tissue che...

General Information

Status
Published
Publication Date
31-Mar-2022
ICS
13.060.70 - Examination of biological properties of water
Technical Committee
E50 - Environmental Assessment, Risk Management and Corrective Action
Drafting Committee
E50.47 - Biological Effects and Environmental Fate
Current Stage
Ref Project

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ASTM E2122-22 - Standard Guide for Conducting In-situ Field Bioassays With Caged BivalvesASTM E2122-22 - Standard Guide for Conducting In-situ Field Bioassays With Caged Bivalves
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ASTM E2122-22 - Standard Guide for Conducting In-situ Field Bioassays With Caged Bivalves
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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: E2122 − 22
Standard Guide for
1
Conducting In-situ Field Bioassays With Caged Bivalves
This standard is issued under the fixed designation E2122; 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 rangeandcompartmentalizedcagesformultiplemeasurements
on the same individuals.
1.1 This guide describes procedures for conducting con-
1.2 The test is referred to as a field bioassay because it is
trolled experiments with caged bivalves under field conditions.
conducted in the field and because it includes an element of
The purpose of this approach is to facilitate the simultaneous
relative chemical potency to satisfy the bioassay definition.
collection of field data to help characterize chemical exposure
Relative potency is established by comparing tissue concen-
and associated biological effects in the same organism under
trations with effects levels for various chemicals with toxicity
environmentally realistic conditions. This approach of charac-
and bioaccumulation end points (6, 7, 8, 9, 10) even though
terizing exposure and effects is consistent with the US EPA
there may be more uncertainty associated with effects mea-
ecological risk assessment paradigm. Bivalves are useful test
surements in field studies. Various pathways of exposure can
organisms for in-situ field bioassays because they (1) concen-
be evaluated because filter-feeding and deposit-feeding are the
trate and integrate chemicals in their tissues and have a more
primary feeding strategies for bivalves. Filter-feeding bivalves
limitedabilitytometabolizemostchemicalsthanotherspecies,
may be best suited to evaluate the bioavailability and associ-
(2) exhibit measurable sublethal effects associated with expo-
atedeffectsofchemicalsinthewatercolumn(thatis,dissolved
suretothosechemicals,(3)providepairedtissuechemistryand
and suspended particulates); deposit-feeding bivalves may be
response data which can be extrapolated to other species and
best suited to evaluate chemicals associated with sediments
trophic levels, (4) provide tissue chemistry data which can be
(11, 12, 13, 14). It may be difficult to demonstrate pathways of
used to estimate chemical exposure from water or sediment,
exposure under field conditions, particularly since filter-
and (5) facilitate controlled experimentation in the field with
feedingbivalvescaningestsuspendedsedimentandfacultative
large sample sizes because they are easy to collect, cage, and
2
deposit-feeding bivalves can switch between filter- and deposit
measure (1, 2) . The experimental control afforded by this
feeding over relatively small temporal scales. Filter-feeding
approach can be used to place a large number of animals of a
bivalves caged within 1 m of bottom sediment have also been
known size distribution in specific areas of concern to quantify
used effectively in sediment assessments from depths of 10 to
exposure and effects over space and time within a clearly
650 m (5, 15, 16). Caged bivalve studies have also been
defined exposure period. Chemical exposure can be estimated
conducted in the intertidal zone (17). The field testing proce-
by measuring the concentration of chemicals in water,
dures described here are useful for testing most bivalves
sediment, or bivalve tissues, and effects can be estimated with
although modifications may be necessary for a particular
survival, growth, and other sublethal end points (3). Although
species.
a number of assessments have been conducted using bivalves
to characterize exposure by measuring tissue chemistry or
1.3 These field testing procedures with caged bivalves are
associated biological effects, relatively few assessments have
applicable to the environmental evaluation of water and
been conducted to characterize both exposure and biological
sediment in marine, estuarine, and freshwater environments
effects simultaneously (2, 4, 5). This guide is specifically
with almost any combination of chemicals, and methods are
designed to help minimize the variability in tissue chemistry
being developed to help interpret the environmental signifi-
and response measurements by using a practical uniform size
cance of accumulated chemicals (6, 7, 9, 18, 19). These
procedures could be regarded as a guide to an exposure system
to assess chemical bioavailability and toxicity under natural,
1 site- specific conditions, where any clinical measurements are
ThisguideisunderthejurisdictionofASTMCommitteeE50onEnvironmental
A
...

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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.  
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SIGNIFICANCE AND USE
5.1 Protection of a species requires the prevention of detrimental effects of chemicals on the survival, growth, reproduction, health, and uses of individuals of that species. Behavioral toxicity tests provide information concerning the sublethal effects of chemicals and signal the presence of toxic test substances.  
5.1.1 The locomotory, feeding, and social responses of fish are adaptive and essential to survival. Major changes in these responses may result in a diminished ability to survive, grow, avoid predation, or reproduce and cause significant changes in the natural population (8). Fish behavioral responses are known to be highly sensitive to environmental variables as well as toxic substances.  
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1.2 Behavioral toxicosis occurs when chemical or other stressful conditions, such as changes in water quality or temperature, induce a behavioral change that exceeds the normal range of variability (1). Behavior includes all of the observable, recordable, or measurable activities of a living organism and reflects genetic, neurobiological, physiological, and environmental determinants (2).  
1.3 Behavioral methods can be used in biomonitoring, in the determination of no-observed-effect and lowest-observed-effect concentrations, and in the prediction of hazardous chemical impacts on natural populations (3).  
1.4 The behavioral methods described in this guide include locomotory activity, feeding, and social responses, which are critical to the survival of fish (4).  
1.5 This guide is arranged as follows:    
Section Number  
Scope  
1  
Referenced Documents  
2  
Terminology  
3  
Summary of Guide  
4  
Significance and Use  
5  
Interferences  
6  
Safety Precautions  
7  
Responses Measured  
8  
Test Organisms  
9  
Facility  
10  
Qualitative Behavioral Assessment Method  
11  
Quantitative Behavioral Measurements  
12  
Experimental Design  
13  
Calculation of Test Results  
14  
Report  
15  
1.6 The values stated in inch-pound units are to be regarded as standard. The values given in parentheses are mathematical conversions to SI units that are provided for information only and are not considered standard. For an explanation of units and symbols, refer to IEEE/ASTM SI 10.  
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ABSTRACT
This guide covers procedures for obtaining laboratory data concerning the adverse effects of aqueous ambient samples and effluents on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, during a short-term exposure, depending on the species, using the static, renewal, and flow-through techniques. These procedures will probably be useful for conducting acute toxicity tests on aqueous effluents with many other aquatic species, although modifications might be necessary. Static tests might not be applicable to effluents that have a high oxygen demand, or contain materials that (1) are highly volatile, (2) are rapidly biologically or chemically transformed in aqueous solutions, or (3) are removed from test solutions in substantial quantities by the test chambers or organisms during the test. Results of acute toxicity tests should usually be reported in terms of a median lethal concentration (LC50) or median effective concentration (EC50). An acute toxicity test does not provide information about whether delayed effects will occur. Specified requirements involving the following are detailed: (1) hazards; (2) apparatus: facilities, special requirements, construction materials, metering system, test chambers, cleaning, and acceptability; (3) dilution water requirements, source, treatment, and characterization; (4) effluent sampling point, collection, preservation, treatment, and test concentrations; (5) test organism species, age, source, care and handling, feeding, disease treatment, holding, acclimation, and quality; (6) procedure: experimental design, dissolved oxygen, temperature, loading, beginning the test, feeding, duration of test, biological data, and other measurements; (7) analytical methodology; (8) acceptability of test; (9) calculation of results; and (1) report of results.
SIGNIFICANCE AND USE
5.1 An acute effluent toxicity test is conducted to obtain information concerning the immediate effects on test organisms of a short-term exposure to an effluent under specific experimental conditions. One can directly examine acute effects of complex mixtures of chemicals as occurs in effluents and some ambient waters. Acute effluent toxicity tests can be used to evaluate the potential for designated-use or aquatic life impairment in the receiving stream, lake, or estuary. 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 effluent 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, (2) toxicity to benthic species might be dependent on sorption or settling of components of the effluent onto the substrate, and (3) the effluent might physically or chemically interact with the receiving water.  
5.3 Results of acute effluent tests might be used to compare the acute sensitivities of different species and the acute toxicities of different effluents, and to study the effects of various environmental factors on results of such tests.  
5.4 Acute tests are usually the first step in evaluating the effects of an effluent on aquatic organisms.  
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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.  
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5.1 Microbiological water testing procedures using membrane filtration are based on the premise that all bacteria within a specific size range will be retained by the membrane filter used. If the membrane filter does not retain these bacteria, false negative results or lowered density estimates may occur that could have serious repercussions due to the presence of unrecognized potential health hazards in the water being tested, especially in drinking water.  
5.2 This procedure as devised will enable the user to test each membrane filter lot number for its ability to retain all bacteria equal to, or larger than, the stated membrane pore size.
SCOPE
1.1 This test method covers a procedure to test membrane filters for their ability to retain bacteria whose diameter is equal to or slightly larger than membrane filters with pore size rated at 0.40 to 0.45 μm.  
1.2 The procedures described are for the use of user laboratories as differentiated from manufacturers’ laboratories.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 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.

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ASTM
ASTM E1611-21(Guide)
Standard Guide for Conducting Sediment Toxicity Tests with Polychaetous Annelids

SIGNIFICANCE AND USE
5.1 The test procedure covered in this guide is not intended to simulate exactly the exposure of benthic polychaetes to chemicals under natural conditions, but rather to provide a conveniently rapid, standard toxicity test procedure yielding a reasonably sensitive indication of the toxicity of materials in marine and estuarine sediments.  
5.2 The protection of a community of organisms requires averting detrimental contaminant-related effects on the number and health of individuals and species within that population. Sediment toxicity tests provide information on the toxicity of test materials in sediments. Theoretically, projection of the most sensitive species within a community will protect the community as a whole.  
5.3 Polychaetes are an important component of the benthic community. They are preyed upon by many species of fish, birds, and larger invertebrate species, and they are predators of smaller invertebrates, larval stages of invertebrates, and, in some cases, algae, as well as organic material associated with sediment. Polychaetes are sensitive to both organic and inorganic chemicals (1, 2).5 The ecological importance of polychaetes, their wide geographical distribution and ability to be cultured in the laboratory, and sensitivity to chemicals, make them appropriate toxicity test organisms.  
5.4 An acute or 10-day toxicity test is conducted to obtain information concerning the immediate effects to a test material on a test organism under specified experimental conditions for a short period of time. An acute toxicity test does not necessarily provide information concerning whether delayed effects will occur, although a post-exposure observation period, with appropriate feeding, if necessary, could provide such information.  
5.5 The results of acute sediment toxicity tests can 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...
SCOPE
1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of potentially contaminated sediment, or of a test material added experimentally to contaminated or uncontaminated sediment, on marine or estuarine infaunal polychaetes during 10-day or 20 to 28-day exposures. These procedures are useful for testing the effects of various geochemical characteristics of sediments on marine and estuarine polychaetes and could be used to assess sediment toxicity to other infaunal taxa, although modifications of the procedures appropriate to the test species might be necessary. Procedures for the 10-day static test are described for Neanthes arenaceodentata and Alitta virens 2 (formerly Nereis virens and Neanthes virens) and for the 20 to 28-day static-renewal sediment toxicity for  N. arenaceodentata.  
1.2 Modifications of these procedures might be appropriate for other sediment toxicity test procedures, such as flow-through or partial life-cycle tests. The methods outlined in this guide should also be useful for conducting sediment toxicity tests with other aquatic taxa, although modifications might be necessary. Other test organisms might include other species of polychaetes, crustaceans, and bivalves.  
1.3 Other modifications of these procedures might be appropriate for special needs or circumstances. Although using appropriate procedures is more important than following prescribed procedures, the results of tests conducted using unusual procedures are not likely to be comparable to those of many other tests. Comparisons of the results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting sediment tests with infaunal organisms.  
1.4 These procedures are applicable to sediments contaminated with most chemicals, either individually or in formulations, commercial products, and known or unknown...

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ASTM
ASTM D3978-21a(Practice)
Standard Practice for Algal Growth Potential Testing with Pseudokirchneriella subcapitata

SIGNIFICANCE AND USE
5.1 The significance of measuring algal growth potential in water samples is that differentiation can be made between the nutrients of a sample determined by chemical analysis and the nutrients that are actually available for algal growth. The addition of nutrients (usually nitrogen and phosphorus singly or in combination) to the sample can give an indication of which nutrient(s) is (are) limiting for algal growth  (1,10,11,12,13,14).
SCOPE
1.1 This practice measures, by Pseudokirchnereilla subcapitata growth response, the biological availability of nutrients, as contrasted with chemical analysis of the components of the sample. This practice is useful for assessing the impact of nutrients, and changes in their loading, upon freshwater algal productivity. Other laboratory or indigenous algae can be used with this practice. However, Pseudokirchnereilla subcapitata must be cultured as a reference alga along with the alternative algal species.  
1.2 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For a specific precautionary statement, see Section 16.  
1.3 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.

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