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ASTM E1218-21

(Guide)

Standard Guide for Conducting Static Toxicity Tests with Microalgae

Standard Guide for Conducting Static Toxicity Tests with Microalgae

SIGNIFICANCE AND USE
5.1 Tests with algae provide information on the toxicity of test materials to an important component of the aquatic biota and might indicate whether additional testing (2) is desirable. Specific testing procedures under various regulatory jurisdictions follow procedures similar to those described in this Guide (3, 4). Users should consult with any specific regulatory requirements to determine the applicability and consistency of this standard with such requirements.  
5.2 Algae are ubiquitous in aquatic ecosystems, where they incorporate solar energy into biomass, produce oxygen, function in nutrient cycling and serve as food for animals. Because of their ecological importance, sensitivity to many toxicants, ready availability, ease of culture, and fast growth rates (rendering it possible to conduct a multi-generation test in a short period of time), algae are often used in toxicity testing.  
5.3 Results of algal toxicity tests might be used to compare the sensitivities of different species of algae and the toxicities of different materials to algae and to study the effects of various environmental factors on results of such tests.  
5.4 Results of algal toxicity tests might be an important consideration when assessing the hazards of materials to aquatic organisms (See Guide E1023) or deriving water quality criteria for aquatic organisms (5).  
5.5 Results of algal toxicity tests might be useful for studying biological availability of, and structure-activity relationships between, test materials.  
5.6 Results of algal toxicity tests will depend on the temperature, composition of the growth medium, and other factors. These tests are conducted in solutions that contain concentrations of salts, minerals, and nutrients that greatly exceed those in most surface waters. These conditions may over- or under-estimate the effects of the test material if discharged to surface waters.
SCOPE
1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of a test material added to growth medium on growth of certain species of freshwater and saltwater microalgae during a static exposure. These procedures will probably be useful for conducting short-term toxicity tests with other species of algae, although modifications might be necessary. Although the test duration is comparable to an acute toxicity test with aquatic animals, an algal toxicity test of short duration (72, 96 or 120 h) allows for examination of effects upon multiple generations of an algal population and thus should not be viewed as an acute toxicity test.  
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 toxicity tests with microalgae.  
1.3 These procedures are applicable to many chemicals, either individually or in formulations, commercial products, or known mixtures. With appropriate modifications, these procedures can be used to conduct tests on temperature, and pH and on such materials as aqueous effluents (see Guide E1192), leachates, oils, particulate matter, sediments, and surface waters. Static tests might not be applicable to materials that are highly volatile, are rapidly biologically or chemically transformed in aqueous solutions, or are removed from test solutions in substantial quantities by the test vessels or organisms during the test. (1)3 However, practical flow-through test procedures with microalgae have not been developed.  
1.4 Results of tests using microalgae should usually be reported in terms of the 96-h (or other time period) IC50 (see 3.2.5) b...

General Information

Status
Published
Publication Date
31-Oct-2021
ICS
07.100.20 - Microbiology 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

Relations

Refers
ASTM D3731-20 - Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters
Effective Date
01-Jun-2020

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ASTM E1218-21 - Standard Guide for Conducting Static Toxicity Tests with MicroalgaeASTM E1218-21 - Standard Guide for Conducting Static Toxicity Tests with Microalgae
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ASTM E1218-21 - Standard Guide for Conducting Static Toxicity Tests with Microalgae
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Standards Content (Sample)

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E1218 − 21
Standard Guide for
1,2
Conducting Static Toxicity Tests with Microalgae
This standard is issued under the fixed designation E1218; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
3
1. Scope during the test. (1) However, practical flow-through test
procedures with microalgae have not been developed.
1.1 This guide covers procedures for obtaining laboratory
1.4 Results of tests using microalgae should usually be
data concerning the adverse effects of a test material added to
growth medium on growth of certain species of freshwater and reported in terms of the 96-h (or other time period) IC50 (see
3.2.5) based on reduction in growth. In some situations, it
saltwater microalgae during a static exposure. These proce-
dures will probably be useful for conducting short-term toxic- might only be necessary to determine whether a specific
concentration unacceptably affects the growth of the test
ity tests with other species of algae, although modifications
mightbenecessary.Althoughthetestdurationiscomparableto species or whether the IC50 is above or below a specific
concentration.
anacutetoxicitytestwithaquaticanimals,analgaltoxicitytest
of short duration (72, 96 or 120 h) allows for examination of
1.5 This guide is arranged as follows:
effects upon multiple generations of an algal population and
Section
thus should not be viewed as an acute toxicity test.
Referenced Documents 2
Terminology 3
1.2 Other modifications of these procedures might be justi-
Summary of Guide 4
fied by special needs or circumstances.Although using appro- Significance and Use 5
Hazards 7
priate procedures is more important than following prescribed
Apparatus 6
procedures,resultsoftestsconductedusingunusualprocedures
Facilities 6.1
are not likely to be comparable to results of many other tests. Equipment 6.2
Test Vessels 6.3
Comparisonofresultsobtainedusingmodifiedandunmodified
Cleaning 6.4
versions of these procedures might provide useful information
Acceptability 6.5
concerning new concepts and procedures for conducting tox- Growth Medium 8
Test Material 9
icity tests with microalgae.
General 9.1
Stock Solution 9.2
1.3 These procedures are applicable to many chemicals,
Test Concentration(s) 9.3
either individually or in formulations, commercial products, or
Test Organisms 10
known mixtures. With appropriate modifications, these proce-
Species 10.1
Source 10.2
dures can be used to conduct tests on temperature, and pH and
Culture 10.3
on such materials as aqueous effluents (see Guide E1192),
Quality 10.4
leachates, oils, particulate matter, sediments, and surface wa-
Procedure 11
Experimental Design 11.1
ters. Static tests might not be applicable to materials that are
Temperature 11.2
highly volatile, are rapidly biologically or chemically trans-
Illumination 11.3
formed in aqueous solutions, or are removed from test solu-
Beginning the Test 11.4
Gas Exchange 11.5
tions in substantial quantities by the test vessels or organisms
Duration of Test 11.6
Biological Data 11.7
Other Measurements 11.8
Determination of Algistatic and Algicidal Effects 11.8.5
1
ThisguideisunderthejurisdictionofASTMCommitteeE50onEnvironmental
Analytical Methodology 12
Assessment, Risk Management and CorrectiveAction and is the direct responsibil-
Acceptability of Test 13
ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.
Calculation 14
Current edition approved Nov. 1, 2021. Published December 2021. Originally
Report 15
approvedin1990.Lastpreviouseditionapprovedin2012asE1218-04(2012)which Keywords 16
was withdrawn January 2021 and reinstated in November 2021. DOI: 10.1520/
E1218-21.
2
This standard guide is a document, developed using the consensus mechanisms
3
of ASTM, that provides guidance for the selection of procedures to accomplish a The boldface numbers given in parentheses refer to a list of references at the
specific test, but which does not stipulate specific procedures. end of the text.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
E1218 − 21
1.6 This standard does not purport to address all of the 3.1.1.3 should—is used to state that the specified condition
safety concerns, if any, associated with its use. It is the is recommended and ought to be met if possible. Although
responsibility of the user of this standard to establish appro- viola
...

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5.1 Polychaetes are an important component of the benthic community, in which they generally comprise 30 to 50 % of the macroinvertebrate population. They are preyed upon by many species of fish, birds, and larger invertebrate species. Larger polychaetes feed on small invertebrates, larval stages of invertebrates, and algae. Polychaetes are especially sensitive to inorganic toxicants and, to a lesser extent, to organic toxicants (1).4 The ecological importance of polychaetes and their wide geographical distribution, ability to be cultured in the laboratory, and sensitivity to contaminants make them appropriate acute and chronic toxicity test organisms. Their relatively short life cycle enables the investigator to measure the effect of contaminants on reproduction.  
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5.1 Ceriodaphnia  was first used as a toxicity test organism by Mount and Norberg (2). Introduced for use in effluent and ambient water evaluations, Ceriodaphnia have also been a valuable addition to single chemical test procedures.  
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SIGNIFICANCE AND USE
5.1 Protection of an aquatic species requires prevention of unacceptable effects on populations in natural habitats. Toxicity tests are conducted to provide data that may be used to predict what changes in numbers and weights of individuals might result from similar exposure to the test material in the natural aquatic environment. Information might also be obtained on the effects of the material on the health of the species.  
5.2 Results of life-cycle tests with D. magna are used to predict chronic effects likely to occur on daphnids in field situations as a result of exposure under comparable conditions.  
5.2.1 Life-cycle tests with D. magna are used to compare the chronic sensitivities of different species, the chronic toxicities of different materials, and study the effects of various environmental factors on the results of such tests.  
5.2.2 Life-cycle tests with D. magna are used to assess the risk of materials to aquatic organisms (see Guide E1023) or derive water quality criteria for aquatic organisms (1).3  
5.2.3 Life-cycle tests with D. magna are used to extrapolate the results of chronic toxicity tests on the same test material with the same species in another water or with another species in the same or a different water. Most such predictions take into account the results of acute toxicity tests, and so the usefulness of the results of a life-cycle test with D. magna may be increased by reporting the results of an acute toxicity test (see Guide E729) conducted under the same conditions. In addition to conducting an acute toxicity test with unfed D. magna, it may be relevant to conduct an acute test in which the daphnids are fed the same as in the life-cycle test to see if the presence of that concentration of that food affects the results of the acute test and the acute-chronic ratio (ACR) (see 10.3.1).  
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SCOPE
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1.2 These procedures are applicable to most chemicals, either individually or in formulations, commercial products, or known mixtures. With appropriate modifications, these procedures can be used to conduct tests on temperature, dissolved oxygen, pH, and on such materials as aqueous effluents (also see Guide E1192), leachates, oils, particulate matter, sediments, and surface waters. The technique, (renewal or flow-through), will be selected based on the chemical characteristics of the test material such as high oxygen demand, volatility, susceptibility to transformation (biologically or chemically), or sorption to glass.  
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5.1 Sediment provides habitat for many aquatic organisms and is a major repository for many of the more persistent chemicals that are introduced into surface waters. In the aquatic environment, most anthropogenic chemicals and waste materials including toxic organic and inorganic chemicals can accumulate in sediment, which can in turn serve as a source of exposure for organisms living on or in sediment. Contaminated sediments may be directly toxic to aquatic life or can be a source of contaminants for bioaccumulation in the food chain.  
5.2 The objective of a sediment test is to determine whether chemicals in sediment are harmful to or are bioaccumulated by benthic organisms. The tests can be used to measure interactive toxic effects of complex chemical mixtures in sediment. Furthermore, knowledge of specific pathways of interactions among sediments and test organisms is not necessary to conduct the tests. Sediment tests can be used to: (1) determine the relationship between toxic effects and bioavailability, (2) investigate interactions among chemicals, (3) compare the sensitivities of different organisms, (4) determine spatial and temporal distribution of contamination, (5) evaluate hazards of dredged material, (6) measure toxicity as part of product licensing or safety testing, (7) rank areas for clean up, and (8) estimate the effectiveness of remediation or management practices.  
5.3 Results of toxicity tests on sediments spiked at different concentrations of chemicals can be used to establish cause and effect relationships between chemicals and biological responses. Results of toxicity tests with test materials spiked into sediments at different concentrations may be reported in terms of a LC50 (median lethal concentration), an EC50 (median effect concentration), an IC50 (inhibition concentration), or as a NOEC (no observed effect concentration) or LOEC (lowest observed effect concentration). However, spiked sediment may not be representative of chemicals a...
SCOPE
1.1 Relevance of Sediment Contamination—Sediment provides habitat for many aquatic organisms and is a major repository for many of the more persistent chemicals that are introduced into surface waters. In the aquatic environment, both organic and inorganic chemicals may accumulate in sediment, which can in turn serve as a source of exposure for organisms living on or in sediment. Contaminated sediments may be directly toxic to aquatic life or can be a source of contaminants for bioaccumulation in the food chain.  
1.2 Sediment Assessment Tools—Several types of information may be useful in assessing the risk, or potential risk, posed by sediment contaminants, including: (1) chemical analysis of sediment contaminants; (2) sediment toxicity tests, (3) bioaccumulation tests; and (4) surveys of benthic community structure. Each of these provides a different type of information to the assessment, and integrating information from all four lines of evidence may often provide the most robust assessments.  
1.3 Strengths of Toxicity Testing of Contaminated Sediments—Directly assessing the toxicity of contaminated sediments provides some of the same advantages to sediment assessment that whole effluent toxicity testing provides to management of industrial and municipal effluents. As for effluent tests, direct testing of sediment toxicity allows the assessment of biological effects even if: (1) the identities of toxic chemicals present are not (or not completely) known; (2) the influence of site-specific characteristics of sediments on toxicity (bioavailability) is not understood; and (3) the interactive or aggregate effects of mixtures of chemicals present are not known or cannot be adequately predicted. In addition, testing the response of benthic or epibenthic organisms exposed via sediment provides an assessment that is based on the same routes of exposure that would exist in nature, rather than only through water column expos...

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ASTM
ASTM D8516-23(Test Method)
Standard Test Method for Quantification of Culturable Waterborne Bacteria Using a Defined Culture Medium Coated Plate

SIGNIFICANCE AND USE
5.1 This plate format is useful for the routine monitoring of culturable, waterborne bacteria in potable and non-potable waters. The significance of finding these bacteria can help with identifying water quality or water system problems or evaluate compliance with maintenance protocols. This test method uses small volumes of water, or dilutions thereof, and provides an easy and reliable method that eliminates media preparation and reduces laboratory waste.
SCOPE
1.1 This test method describes a simple procedure for the quantification of culturable, waterborne bacteria in potable water (drinking water, bottled water, and dental water, for example) and non-potable waters (cooling towers, for example).  
1.1.1 The EasyDisc2, 3 plate format is designed to test 1 mL of a water sample on a 47 mm gridded plate containing a growth reagent embedded to the plate’s inner surface.  
1.1.2 Detection is based on colorimetric technology in which viable, aerobic, heterotrophic, waterborne bacteria grow when present in the water sample, displaying a color reaction which allows for a simplified visualization of colony growth.  
1.2 Each plate can accurately detect up to 300 colony forming units per 1 mL (CFU/1 mL) of sample. To increase the quantification range, a sample dilution can be used. Adjust the CFU/mL result to reflect dilutions.  
1.3 This test method can be used for potable (for example, drinking, bottled, and dental) waters and non-potable waters such as cooling tower waters. It is the user’s responsibility to adhere to all requirements by local regulations and ensure the validity of this test method for waters other than those tested as part of the Interlaboratory Study (ILS).  
1.4 The values stated in SI units are to be regarded as the standard. No other units of measurement are included in this standard.  
1.5 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.  
1.6 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 E1023-23(Guide)
Standard Guide for Assessing the Hazard of a Material to Aquatic Organisms and Their Uses

SIGNIFICANCE AND USE
5.1 Adverse effects on natural populations of aquatic organisms and their uses have demonstrated the need to assess the hazards of many new, and some presently used, materials. The process described herein will help producers, users, regulatory agencies, and others to efficiently and adequately compare alternative materials, completely assess a final candidate material, or reassess the hazard of a material already in use.  
5.2 Sequential assessment and feedback allow appropriate judgments concerning efficient use of resources, thereby minimizing unnecessary testing and focusing effort on the information most pertinent to each material. For different materials and situations, assessment of hazard will appropriately be based on substantially different amounts and kinds of biological, chemical, physical, and toxicological data.  
5.3 Assessment of the hazard of a material to aquatic organisms and their uses should never be considered complete for all time. Reassessment should be considered if the amount of production, use, or disposal increases, new uses are discovered, or new information on biological, chemical, physical, or toxicological properties becomes available. Periodic review will help assure that new circumstances and information receive prompt appropriate attention.  
5.4 If there is substantial transformation to another material, the hazard of both materials may need to be assessed.  
5.5 In many cases, consideration of adverse effects should not end with completion of the hazard assessment. Additional steps should often include risk assessment, decisions concerning acceptability of identified hazards and risks, and mitigative actions.  
5.6 Because this practice deals mostly with adverse effects on aquatic organisms and their uses, it is important that mitigative actions, such as improved treatment of aqueous effluents, not result in unacceptable effects on non-aquatic organisms. Thus, this standard should be used with other information in order to a...
SCOPE
1.1 This guide describes a stepwise process for using information concerning the biological, chemical, physical, and toxicological properties of a material to identify adverse effects likely to occur to aquatic organisms and their uses as a result of release of the material to the environment. The material will usually be a specific chemical, although it might be a group of chemicals that have very similar biological, chemical, physical, and toxicological properties and are usually produced, used, and discarded together.  
1.2 The hazard assessment process is complex and requires decisions at a number of points; thus, the validity of a hazard assessment depends on the soundness of those decisions, as well as the accuracy of the information used. All decisions should be based on reasonable worst-case analyses so that an appropriate assessment can be completed for the least cost that is consistent with scientific validity.  
1.3 This guide assumes that the reader is knowledgeable in aquatic toxicology and related pertinent areas. A list of general references is provided (1).2  
1.4 This guide does not describe or reference detailed procedures for estimating or measuring environmental concentrations, or procedures for determining the maximum concentration of test material that is acceptable in the food of predators of aquatic life. However, this guide does describe how such information should be used when assessing the hazard of a material to aquatic organisms and their uses.  
1.5 Because assessment of hazard to aquatic organisms and their uses is a relatively new activity within aquatic toxicology, most of the guidance provided herein is qualitative rather than quantitative. When possible, confidence limits should be calculated and taken into account.  
1.6 This guide provides guidance for assessing hazard but does not provide guidance on how to take into account social considerations in order to judge the acceptabil...

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ASTM
ASTM E2783-22(Test Method)
Standard Test Method for Assessment of Antimicrobial Activity for Water Miscible Compounds Using a Time-Kill Procedure

SIGNIFICANCE AND USE
5.1 This procedure may be used to assess the in vitro reduction of a microbial population of test organisms after exposure to a test material.
SCOPE
1.1 This test method measures the changes of a population of aerobic and anaerobic microorganisms within a specific sampling time when tested against antimicrobial test materials in vitro. The organisms used are standardized as to growth requirements and inoculum preparation and must grow under the conditions of the test. The primary purpose of this test method is to provide a set of standardized conditions and test organisms to facilitate comparative assessments of antimicrobial materials miscible in aqueous systems.  
1.2 This test method allows the option of using a test sample size of 10 mL or 100 mL.  
1.3 Knowledge of microbiological techniques is required for this procedure.  
1.4 Aseptic technique should be practiced at all times.  
1.5 In this test method, SI units are used for all applications, except for distance in which case inches are used and SI units follow in parentheses.  
1.6 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.  
1.7 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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