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ASTM D5864-95

(Test Method)

Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components

Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components

SCOPE
1.1 This test method covers the determination of the degree of aerobic aquatic biodegradation of fully formulated lubricants or their components on exposure to an inoculum under laboratory conditions.  
1.2 This test method is intended to specifically address the difficulties associated with testing water insoluble materials and complex mixtures such as are found in many lubricants.  
1.3 This test method is designed to be applicable to all lubricants that are not volatile and are not inhibitory at the test concentration to the organisms present in the inoculum.  
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 and health practices and determine the applicability of regulatory limitations prior to use. Specific hazards are discussed in Section 10.

General Information

Status
Historical
Publication Date
09-Dec-2000
ICS
13.060.70 - Examination of biological properties of water
Technical Committee
D02 - Petroleum Products, Liquid Fuels, and Lubricants
Drafting Committee
D02.12 - Environmental Standards for Lubricants
Current Stage
Ref Project

Relations

Replaced By
ASTM D5864-00 - Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components
Effective Date
10-Dec-2000

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ASTM D5864-95 - Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their ComponentsASTM D5864-95 - Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components
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ASTM D5864-95 - Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components
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Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or
withdrawn. Contact ASTM International (www.astm.org) for the latest information.
Designation: D 5864 – 95
AMERICAN SOCIETY FOR TESTING AND MATERIALS
100 Barr Harbor Dr., West Conshohocken, PA 19428
Reprinted from the Annual Book of ASTM Standards. Copyright ASTM
Standard Test Method for
Determining Aerobic Aquatic Biodegradation of Lubricants
1
or Their Components
This standard is issued under the fixed designation D 5864; 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 (e) indicates an editorial change since the last revision or reapproval.
6
1. Scope 2.3 APHA Standard:
2540B Total Solids Dried at 103–105°C
1.1 This test method covers the determination of the degree
9215 Heterotrophic Plate Count
of aerobic aquatic biodegradation of fully formulated lubri-
cants or their components on exposure to an inoculum under
3. Terminology
laboratory conditions.
3.1 Definitions:
1.2 This test method is intended to specifically address the
3.1.1 Definitions of terms applicable to this test method that
difficulties associated with testing water insoluble materials
are not described herein appear in the Compilation of ASTM
and complex mixtures such as are found in many lubricants.
7
Standard Definitions, 1990 or Terminology E 943.
1.3 This test method is designed to be applicable to all
3.1.2 aerobic, adj—(1) taking place in the presence of
lubricants that are not volatile and are not inhibitory at the test
oxygen, (2) living or active in the presence of oxygen.
concentration to the organisms present in the inoculum.
3.1.3 biodegradation, n—the process of chemical break-
1.4 This standard does not purport to address all of the
down or transformation of a substance caused by organisms or
safety concerns, if any, associated with its use. It is the
their enzymes.
responsibility of the user of this standard to establish appro-
3.1.4 biomass, n—any material, excluding fossil fuels,
priate safety and health practices and determine the applica-
which is or was a living organism or component of a living
bility of regulatory limitations prior to use. Specific hazards are
organism.
discussed in Section 10.
3.1.5 blank, n—a flask containing the test medium and the
inoculum with no additional carbon source added.
2. Referenced Documents
3.1.6 inoculum, n—spores, bacteria, single celled organ-
isms, or other live materials, that are introduced into a test
2.1 ASTM Standards:
2 medium.
D 1129 Terminology Relating to Water
2 3.1.7 lag phase, n—the period of physiological activity and
D 1193 Specification for Reagent Water
2 diminished cell division following the addition of microorgan-
D 1293 Test Methods for pH of Water
8
isms to a new culture medium.
D 4447 Guide for Disposal of Laboratory Chemicals and
3
3.1.8 log phase, n—the period of growth of microorganisms
Samples
8
during which cells divide at a constant rate.
D 5291 Test Methods for Instrumental Determination of
3.1.9 mixed liquor, n—the contents of an aeration tank
Carbon, Hydrogen, and Nitrogen in Petroleum Products
4 including the activated sludge mixed with primary effluent or
and Lubricants
the raw wastewater and return sludge.
E 943 Terminology Relating to Biological Effects and En-
3 3.1.10 pre-adaptation, n—the incubation of an inoculum in
vironmental Fate
the presence of the test substance which is done prior to the
5
2.2 ISO Standard:
initiation of the test and under conditions similar to the test
4259:1992(E) Petroleum Products—Determination and Ap-
conditions.
plication of Precision Data in Relation to Methods of Test
3.1.10.1 Discussion—The aim of pre-adaptation is to im-
prove the precision of the test method by decreasing variability
in the rate of biodegradation produced by the inoculum.
Pre-adaptation may mimic the natural processes which cause
1
This test method is under the jurisdiction of ASTM Committee D-2 on
Petroleum Products and Lubricantsand is the direct responsibility of Subcommittee
D02.12on Environmental Standards for Lubricants.
6
Current edition approved Dec. 10, 1995. Published February 1996. From Standard Methods for the Examination of Water and Wastewater, latest
2
Annual Book of ASTM Standards, Vol 11.01. edition. Available from the American Public Health Association, 1015 18th St.,
3
Annual Book of ASTM Standards, Vol 11.04. N.W., Washington, DC 20036.
4 7
Annual Book of ASTM Standards, Vol 05.03. Available from ASTM Headquarters.
5 8
Available from American National Standards Institute, 11 West 42nd St., New Adapted from McGraw-Hill Dictionary of Scientific and Technical Terms, 4th
York, NY, 10036. ed., 1989.
1

---------------------- Page: 1 ----------------------
D 5864
changes in the microbial population of the inoculum leading to 5.4 Activated sewage-sludge from a sewage-treatmen
...

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ASTM
ASTM D5864-23(Test Method)
Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components

SIGNIFICANCE AND USE
5.1 Results from the test method suggest, within the confines of a controlled laboratory setting, the degree of aerobic aquatic biodegradation of a lubricant or components of a lubricant by measuring the evolved carbon dioxide upon exposure of the test material to an inoculum. The plateau level of CO2 evolution in this test method will suggest the degree of biodegradability of the lubricant. Test substances that achieve a high degree of biodegradation in this test may be assumed to easily biodegrade in many aerobic aquatic environments.  
5.2 Because of the stringency of this test, a low yield of CO2 does not necessarily mean that the test substance is not biodegradable under environmental conditions, but indicates that further testing is necessary to establish biodegradability.  
5.3 Information on toxicity to the inoculum of the test substance may be useful in the interpretation of low biodegradation results.  
5.4 Activated sewage-sludge from a sewage-treatment plant that principally treats domestic waste is considered an acceptable active aerobic inoculum available over a wide geographical area in which to test a broad range of lubricants. An inoculum derived from soil or natural surface waters, or both, or any combination of the three sources, is also appropriate for this test method.
Note 1: Allowance for various and multiple inoculum sources provides access to a greater diversity of biochemical competency and potentially represents more accurately the capacity for biodegradation.  
5.5 A reference or control substance known to biodegrade is necessary in order to verify the activity of the inoculum. The test must be regarded as invalid and should be repeated using a fresh inoculum if the reference does not demonstrate a biodegradation of >60 % of the theoretical CO2 evolution within 28 days.  
5.6 A total CO2 evolution in the blank at the end of the test exceeding 75 mg CO2 per 3 L of medium shall be considered as invalidating the test.  
5.7 The water...
SCOPE
1.1 This test method covers the determination of the degree of aerobic aquatic biodegradation of fully formulated lubricants or their components on exposure to an inoculum under laboratory conditions.  
1.2 This test method is intended to specifically address the difficulties associated with testing water insoluble materials and complex mixtures such as are found in many lubricants.  
1.3 This test method is designed to be applicable to all lubricants that are not volatile and are not inhibitory at the test concentration to the organisms present in the inoculum.  
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. Specific hazards are discussed in Section 10.  
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 D5864-23(Test Method)
Standard Test Method for Determining Aerobic Aquatic Biodegradation of Lubricants or Their Components

SIGNIFICANCE AND USE
5.1 Results from the test method suggest, within the confines of a controlled laboratory setting, the degree of aerobic aquatic biodegradation of a lubricant or components of a lubricant by measuring the evolved carbon dioxide upon exposure of the test material to an inoculum. The plateau level of CO2 evolution in this test method will suggest the degree of biodegradability of the lubricant. Test substances that achieve a high degree of biodegradation in this test may be assumed to easily biodegrade in many aerobic aquatic environments.  
5.2 Because of the stringency of this test, a low yield of CO2 does not necessarily mean that the test substance is not biodegradable under environmental conditions, but indicates that further testing is necessary to establish biodegradability.  
5.3 Information on toxicity to the inoculum of the test substance may be useful in the interpretation of low biodegradation results.  
5.4 Activated sewage-sludge from a sewage-treatment plant that principally treats domestic waste is considered an acceptable active aerobic inoculum available over a wide geographical area in which to test a broad range of lubricants. An inoculum derived from soil or natural surface waters, or both, or any combination of the three sources, is also appropriate for this test method.
Note 1: Allowance for various and multiple inoculum sources provides access to a greater diversity of biochemical competency and potentially represents more accurately the capacity for biodegradation.  
5.5 A reference or control substance known to biodegrade is necessary in order to verify the activity of the inoculum. The test must be regarded as invalid and should be repeated using a fresh inoculum if the reference does not demonstrate a biodegradation of >60 % of the theoretical CO2 evolution within 28 days.  
5.6 A total CO2 evolution in the blank at the end of the test exceeding 75 mg CO2 per 3 L of medium shall be considered as invalidating the test.  
5.7 The water...
SCOPE
1.1 This test method covers the determination of the degree of aerobic aquatic biodegradation of fully formulated lubricants or their components on exposure to an inoculum under laboratory conditions.  
1.2 This test method is intended to specifically address the difficulties associated with testing water insoluble materials and complex mixtures such as are found in many lubricants.  
1.3 This test method is designed to be applicable to all lubricants that are not volatile and are not inhibitory at the test concentration to the organisms present in the inoculum.  
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. Specific hazards are discussed in Section 10.  
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 E1192-23(Guide)
Standard Guide for Conducting Acute Toxicity Tests on Aqueous Ambient Samples and Effluents with Fishes, Macroinvertebrates, and Amphibians

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.  
5.5 Results of acute effluent tests will depend on the temperature, composition of the dilution water, condition of the test organisms, exposure technique, and other factors.
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1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of an aqueous effluent on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, usually during 2 day to 4 day exposures, depending on the species, using the static, renewal, and flow-through techniques. These procedures will probably be useful for conducting acute toxicity tests on ...

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ASTM E2122-22(Guide)
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...
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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...

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ASTM E2591-22(Guide)
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...
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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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ASTM D3863-22(Test Method)
Standard Test Method for Retention Characteristics of 0.40 to 0.45 μm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality

SIGNIFICANCE AND USE
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 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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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 E1711-20(Guide)
Standard Guide for Measurement of Behavior During Fish Toxicity Tests

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.  
5.2 Results from behavioral toxicity tests may be useful for measuring injury resulting from the release of hazardous materials (9).  
5.3 Behavioral responses can also be qualitatively assessed in a systematic manner during toxicity tests to discern trends in sublethal contaminant effects (5).  
5.4 The assessment of locomotory, feeding, and social behaviors is useful for monitoring effluents and sediments from contaminated field sites as well as for defining no-effect concentrations in the laboratory or under controlled field conditions. Such behavioral modifications provide an index of sublethal toxicity and also indicate the potential for subsequent mortality.  
5.5 Behavioral toxicity data can be used to predict the effects of exposure likely to occur in the natural environment (10).  
5.6 Results from behavioral toxicity tests might be an important consideration when assessing the hazard of materials to aquatic organisms. Such results might also be used when deriving water quality criteria for fish and aquatic invertebrate organisms.  
5.7 Results from behavioral toxicity tests can be used to compare the sensitivities of different species, the relative toxicity of different chemical substances on the same organism, or the effect of various environmental...
SCOPE
1.1 This guide covers some general information on methods for qualitative and quantitative assessment of the behavioral responses of fish during standard laboratory toxicity tests to measure the sublethal effects of exposure to chemical substances. This guide is meant to be an adjunct to toxicity tests and should not interfere with those test procedures.  
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.  
1.7 This standard does ...

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ASTM
ASTM D5259-19(Test Method)
Standard Test Method for Isolation and Enumeration of Enterococci from Water by the Membrane Filter Procedure

SIGNIFICANCE AND USE
5.1 The enterococci are indicators of the bacteriological quality for potable water, shellfish growing waters, ambient, and recreational waters. A direct relationship between swimming, associated gastroenteritis, and enterococci has been established through epidemiological studies and marine and fresh water bathing beaches. These studies have led to the development of criteria that can be used to establish bathing water standards based on established health-water quality relationships.  
5.2 Since small or large volumes of water or dilutions thereof, can be analyzed by the membrane filter technique, a wide range of levels of enterococci in water can be enumerated and detected.
SCOPE
1.1 This test method covers a membrane filter (MF) procedure for the detection and enumeration of the enterococci bacteria in water. The enterococci, which include Entero-coccus faecalis (E. faecalis), E. faecium, and their varieties are commonly found in the feces of humans and other warm-blooded animals. Although some strains are ubiquitous and not related to fecal pollution, enterococci in water are an indication of fecal pollution and the possible presence of enteric pathogens. These bacteria are found in water and wastewater in a wide range of densities. The detection limit is one colony forming unit (CFU)/volume filtered.  
1.2 This test method has been used successfully with temperate fresh and marine ambient waters, and wastewaters. It is the user’s responsibility to ensure the validity of this test method for waters of untested types.  
1.3 The values stated in SI units are to be regarded as standard. The values given in parentheses after SI units are included for information only and are not considered 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. For specific hazard statements, see Section 9.  
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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