This document specifies a method for the enumeration of intestinal enterococci in water, including Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus avium, Enterococcus gallinarum, Enterococcus hirae, Enterococcus casselifavus. The method is based on the growth of target organisms in a liquid medium and calculation of the “most probable number” (MPN) of microorganisms by reference to MPN tables or using suitable MPN informatic programs.
This method can be applied to drinking water and bathing water (fresh or marine), together with other similar water types including those containing an appreciable amount of suspended matter, and allows the detection of enterococci at 1 colony-forming unit (CFU) per 100 ml with definitive results within (26 ± 2) h in the presence of heterotrophic bacteria in numbers as high as 1 × 106 per 100 ml of sample.
For bathing waters, fresh and marine, enterococci are best enumerated when samples are diluted 1:10.
The test specified in this document relies upon the detection of intestinal enterococci based upon expression of the enzyme ß-D-glucosidase and provides a confirmed result in 24 h without further testing of positive wells.
This document does not apply to bottled waters, for which the method has not been validated and therefore is outside the scope of this document, unless appropriate validation of performance of this method has been undertaken by the laboratory prior to use.

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This document specifies a method for the enumeration of intestinal enterococci in water, including Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus avium, Enterococcus gallinarum, Enterococcus hirae, Enterococcus casselifavus. The method is based on the growth of target organisms in a liquid medium and calculation of the “most probable number” (MPN) of microorganisms by reference to MPN tables or using suitable MPN informatic programs.
This method can be applied to drinking water and bathing water (fresh or marine), together with other similar water types including those containing an appreciable amount of suspended matter, and allows the detection of enterococci at 1 colony-forming unit (CFU) per 100 ml with definitive results within (26 ± 2) h in the presence of heterotrophic bacteria in numbers as high as 1 × 106 per 100 ml of sample.
For bathing waters, fresh and marine, enterococci are best enumerated when samples are diluted 1:10.
The test specified in this document relies upon the detection of intestinal enterococci based upon expression of the enzyme ß-D-glucosidase and provides a confirmed result in 24 h without further testing of positive wells.
This document does not apply to bottled waters, for which the method has not been validated and therefore is outside the scope of this document, unless appropriate validation of performance of this method has been undertaken by the laboratory prior to use.

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This document specifies a method for the enumeration of intestinal enterococci in water, including Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus avium, Enterococcus gallinarum, Enterococcus hirae, Enterococcus casselifavus. The method is based on the growth of target organisms in a liquid medium and calculation of the “most probable number” (MPN) of microorganisms by reference to MPN tables or using suitable MPN informatic programs. This method can be applied to drinking water and bathing water (fresh or marine), together with other similar water types including those containing an appreciable amount of suspended matter, and allows the detection of enterococci at 1 colony-forming unit (CFU) per 100 ml with definitive results within (26 ± 2) h in the presence of heterotrophic bacteria in numbers as high as 1 × 106 per 100 ml of sample. For bathing waters, fresh and marine, enterococci are best enumerated when samples are diluted 1:10. The test specified in this document relies upon the detection of intestinal enterococci based upon expression of the enzyme ß-D-glucosidase and provides a confirmed result in 24 h without further testing of positive wells. This document does not apply to bottled waters, for which the method has not been validated and therefore is outside the scope of this document, unless appropriate validation of performance of this method has been undertaken by the laboratory prior to use.

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ISO 10705-3:2003 specifies the general principles for assessing the performance of methods for the concentration of bacteriophages from water. Concentration is recommended for those water samples expected to contain < 3 pfp (plaque-forming particles) per millilitre. Concentration methods can be applied to all kinds of water provided that the amount and nature of suspended solids and/or dissolved matter do not interfere with the concentration procedure.
ISO 10705-3:2003 does not give specific details of concentration methods, but outlines the fundamental principles for evaluating the suitability of a particular method for a given type and volume of water. Annex A gives examples of methods that have been found satisfactory and their fields of application.

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ISO 10705-3:2003 specifies the general principles for assessing the performance of methods for the concentration of bacteriophages from water. Concentration is recommended for those water samples expected to contain < 3 pfp (plaque-forming particles) per millilitre. Concentration methods can be applied to all kinds of water provided that the amount and nature of suspended solids and/or dissolved matter do not interfere with the concentration procedure.
ISO 10705-3:2003 does not give specific details of concentration methods, but outlines the fundamental principles for evaluating the suitability of a particular method for a given type and volume of water. Annex A gives examples of methods that have been found satisfactory and their fields of application.

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This document provides the guidelines, minimum requirements and performance characteristics intended to guarantee that manufactured systems intended for on-site/field use (i.e. outside the laboratory) provide reliable and reproducible results. This document specifies the requirements for technologies that enable on-site detection and quantification of Legionella spp. and L. pneumophila using a quantitative polymerase chain reaction assay (qPCR). It specifies general methodological requirements, performance evaluation requirements and quality control requirements. This document is intended to be used by manufacturers of these technologies so that they produce detection systems that end users can operate safely and effectively. End users will be guided by this document to adhere to manufacturer’s instructions, to ensure user competency and to perform the necessary controls. Technical details specified in this document are given for information only. Any other technical solutions complying with the performance requirements are suitable. NOTE For validation and performance requirements, see Clause 9. This document is intended to be applied in the bacteriological investigation of all types of water (hot or cold water, cooling tower water, etc.), unless the nature and/or content of suspended matter and/or background microorganisms interfere with the determination. This interference can result in an adverse effect on both the detection limit and the quantification limit. The results are expressed as the number of genome units of Legionella spp. and/or L. pneumophila per millilitre (or litre) of sample. Although the method described in this document is applicable to all types of water, some additives, such as chemicals used for water treatment, can interfere with and/or affect the sensitivity of the method. The qPCR methods do not give any information about the physiological state of the Legionella. However, there are on-site qPCR methodologies which are able to distinguish intact bacteria from free DNA.

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SIGNIFICANCE AND USE
5.1 This test method is useful for measuring recreational water quality and chlorinated wastewaters, although it can be used for any water suspected of contamination by fecal wastes of warm-blooded animals. The significance of finding E. coli in recreational water samples, especially samples obtained from fresh recreational waters, is that there is a risk of gastrointestinal illness, directly related to the E. coli density, associated with swimming.5  
5.2 Since small or large volumes of water or dilutions thereof can be analyzed by the MF technique, a wider range of levels of E. coli in water can be detected and enumerated than with other methods.
SCOPE
1.1 This test method describes a membrane filter (MF) procedure for the detection and enumeration of Escherichia coli, a bacterium found exclusively in the feces of humans and other warm-blooded animals. The presence of these microorganisms in water is 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 of this procedure is one colony forming unit (CFU) per 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 other types.  
1.3 The values stated in SI units are to be regarded as 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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SIGNIFICANCE AND USE
5.1 P. aeruginosa is an opportunistic pathogen and has been linked as the causative agent of numerous infections that may be transmitted through a contaminated water supply to a susceptible host.
Note 1: Fecal waste is >95 % E. coli which is found in humans and warm blooded animals.  
5.2 The membrane filtration procedure described is a rapid and reliable test method of detecting P. aeruginosa in water.
SCOPE
1.1 The test method covers the isolation and enumeration of Pseudomonas aeruginosa. Testing was performed on spiked samples using reagent grade water as the diluent from surface waters; recreational waters; ground water, water supplies; especially rural nonchlorinated sources; waste water; and saline waters. The detection limit of this test method is one microorganism per 100 mL.  
1.2 This test method was used successfully with reagent water. It is the user's responsibility to ensure the validity of this test method for surface waters, recreational waters, ground water, rural nonchlorinated sources; waste water; and saline waters.  
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. Specific hazard statements are given 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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SIGNIFICANCE AND USE
5.1 Sulfate-reducing bacteria are widely distributed in marine and fresh water muds which, in consequence, frequently are laden with the hydrogen sulfide produced by these organisms during dissimilatory sulfate reduction.  
5.2 It has been reported that Desulfovibrio spp. can form as much as 10 g of sulfide per litre during active multiplication. Sulfate-reducing bacteria can cause the external or internal corrosion of water or wastewater pipelines and pipelines for petroleum and natural gas. The formation of galvanic cells by massive growth of sulfate-reducing bacteria under suitable conditions makes the corrosion much worse than just the effect of the hydrogen sulfide on the metal or concrete.
SCOPE
1.1 These test methods cover the procedure for the detection and enumeration by the most probable number (MPN) technique of sulfate-reducing bacteria in water or water-formed deposits.  
1.2 Two media preparations are provided. Medium A which is prepared with reagent grade water, and Medium B which is prepared using the water to be sampled as the water source. Medium B is offered for those special conditions where sulfate-reducing bacterial strains have adapted to atypical non-fresh water environment.  
1.3 For the isolation and enumeration of thermophilic sulfate-reducing bacteria encountered in waters associated with oil and gas production, all broths, dilution blanks, and incubations must be maintained at temperatures of at least 45 °C and preferably within 5 °C at the sample temperature.  
1.4 The sensitivity of these test methods can be increased by purging the dilution blanks and tubes of media with nitrogen immediately prior to use.  
1.5 The analyst should be aware that adequate collaborative data for precision and bias statements as required by Practice D2777 are not provided. See Section 11 for details.  
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.7 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.  
1.8 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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SIGNIFICANCE AND USE
5.1 This test provides an easy and reliable method for the detection of enterococci in water within 24 h. For recreational water (fresh and marine) testing is performed to insure areas are safe for swimming. Enterolert also can be used for testing bottled water, wastewater, ground water, and drinking water.
SCOPE
1.1 This test method covers a simple procedure for the detection of enterococci in water and wastewater. It is based on IDEXX’s patented Defined Substrate Technology (DST).2 This product, Enterolert, utilizes a nutrient indicator that fluoresces when metabolized. It can detect these bacteria at one most probable number (MPN)/100 mL within 24 h. The presence of this microorganism in water is an indication of fecal contamination and the possible presence of enteric pathogens.  
1.2 This test method can be used successfully with drinking water, source water, recreational (fresh and marine) water, wastewater, and bottled water. It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.  
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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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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This document specifies the requirements for the performance testing of membrane filters used for the retention followed by direct enumeration of microorganisms by culture methods.
This document is applicable to membrane filters which are used for retention followed by direct enumeration of specific microorganisms on solid media or on other devices containing media, like absorbent pads[19].
This document is not applicable for membrane filters used for concentration and elution or for qualitative methods.
These tests are applicable to the membrane filters intended for the microbiological analysis of different types of water, such as:
—    drinking water, bottled water and other types of water with expected low numbers of microorganisms;
—    water with expected higher numbers of microorganisms, for example, surface water and process water.
These tests are intended to demonstrate the suitability of the whole system (membrane filter together with the culture medium including the filtration step) required for the specific tests described in References [3], [6], [8], [10], [12] and [13].
This document applies to:
—    manufacturers producing membrane filters;
—    microbiological laboratories using membrane filters for their own testing or providing these to other end users.

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This document specifies the requirements for the performance testing of membrane filters used for the retention followed by direct enumeration of microorganisms by culture methods.
This document is applicable to membrane filters which are used for retention followed by direct enumeration of specific microorganisms on solid media or on other devices containing media, like absorbent pads[19].
This document is not applicable for membrane filters used for concentration and elution or for qualitative methods.
These tests are applicable to the membrane filters intended for the microbiological analysis of different types of water, such as:
—    drinking water, bottled water and other types of water with expected low numbers of microorganisms;
—    water with expected higher numbers of microorganisms, for example, surface water and process water.
These tests are intended to demonstrate the suitability of the whole system (membrane filter together with the culture medium including the filtration step) required for the specific tests described in References [3], [6], [8], [10], [12] and [13].
This document applies to:
—    manufacturers producing membrane filters;
—    microbiological laboratories using membrane filters for their own testing or providing these to other end users.

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This document specifies the requirements for the performance testing of membrane filters used for the retention followed by direct enumeration of microorganisms by culture methods. This document is applicable to membrane filters which are used for retention followed by direct enumeration of specific microorganisms on solid media or on other devices containing media, like absorbent pads[19]. This document is not applicable for membrane filters used for concentration and elution or for qualitative methods. These tests are applicable to the membrane filters intended for the microbiological analysis of different types of water, such as: - drinking water, bottled water and other types of water with expected low numbers of microorganisms; - water with expected higher numbers of microorganisms, for example, surface water and process water. These tests are intended to demonstrate the suitability of the whole system (membrane filter together with the culture medium including the filtration step) required for the specific tests described in References [3], [6], [8], [10], [12] and [13]. This document applies to: - manufacturers producing membrane filters; - microbiological laboratories using membrane filters for their own testing or providing these to other end users.

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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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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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This document specifies a horizontal method for the enumeration of coagulase-positive staphylococci by counting the colonies obtained on a solid medium (rabbit plasma fibrinogen agar medium) after aerobic incubation at 34 °C to 38 °C (see Reference [10]).
This document is applicable to:
—    products intended for human consumption;
—    products intended for animal feeding;
—    environmental samples in the area of food and feed production and handling;
—    samples from the primary production stage.
This horizontal method was originally developed for the examination of all samples belonging to the food chain.
Because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
Based on the information available at the time of publication of this document, this method is particularly suitable for the examination of fermented products or other products containing technological microbiota based on Staphylococcus spp. (e.g. S. xylosus) (such as cheeses made from raw milk and certain raw meat products) likely to be contaminated by:
—     staphylococci forming atypical colonies on a Baird-Parker agar medium;
—     background microbiota that can obscure the colonies being sought.
Nevertheless, both ISO 6888-1 and this document are given equivalent status.

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SIGNIFICANCE AND USE
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.  
5.2 Protection of a population requires prevention of unacceptable effects on the number, weight, health, and uses of the individuals of that species, or species for which the test species serves as a surrogate. A three-brood toxicity test is conducted to help determine changes in survival and the number of neonates produced that result from exposure to the test material.  
5.3 Results of three-brood toxicity tests with C. dubia might be used to predict chronic or partial chronic effects on species in field situations as a result of exposure under comparable conditions.  
5.4 Results of three-brood toxicity tests with C. dubia might be compared with the chronic sensitivities of different species and the chronic toxicities of different materials, and to study the effects of various environmental factors on results of such tests.  
5.5 Results of three-brood toxicity tests with C. dubia might be useful for predicting the results of chronic 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 are based on the results of acute toxicity tests, and so the usefulness of the results of a three-brood toxicity test with C. dubia might be greatly increased by also reporting the results of an acute toxicity test (see Guides E729 and E1192) conducted under the same conditions. In addition to conducting an acute test with unfed C. dubia, it might also be desirable to conduct an acute test in which the organisms are fed the same as in the three-brood test, to see if the presence of that concentration of that food affects the results of the acute test and the acute chronic ratio (see 10.4.1).  
5.5.1 A 48 or 96-h EC50 or LC50 can sometimes be obtaine...
SCOPE
1.1 This guide describes procedures for obtaining data concerning the adverse effects of an effluent or a test material (added to dilution water, but not to food) on Ceriodaphnia dubia Richard 1894, during continuous exposure throughout a portion of the organism's life. These procedures should also be useful for conducting life cycle toxicity tests with other Cladocera (Guide E1193), although modifications will be necessary.  
1.2 These procedures are applicable to most chemicals, either individually or in formulations, commercial products, or known mixtures, that can be measured accurately at the necessary concentrations in water. With appropriate modifications these procedures can be used to conduct tests on temperature, dissolved oxygen, pH, dissolved ions, and on such materials as aqueous effluents (see also Guide E1192), leachates, oils, particulate matter, sediments (see also Guide E1706), and surface waters. Renewal tests might not be applicable to materials that have high oxygen demand, are highly volatile, are rapidly biologically or chemically transformed, or sorb to test chambers. If the concentration of dissolved oxygen falls below 4 mg/L or the concentration of test material decreases by more than 20 % in test solution(s) at any concentration between renewals, more frequent renewals might be necessary.  
1.3 Other modifications of these procedures might be justified by special needs or circumstances. Results of tests conducted using unusual procedures are not likely to be comparable to results of many other tests. Comparisons of results obtained using modified and unmodified versions of these procedures might provide useful information on new concepts and procedures for conducting three-brood toxicity tests with C. dubia.  
1.4 This guide is arranged as follows:    
Section  
Referenced Documents  
2  
Terminology  
3  
Summary of Guide  
4  
Significance and Use  
5  
Apparatus ...

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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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SIGNIFICANCE AND USE
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.  
5.2 An acute toxicity or chronic text is conducted to obtain information concerning the immediate effects of an exposure to a test material on a test organism under specified experimental conditions. An acute toxicity test provides data on the short-term effects, which are useful for comparisons to other species but do not provide information on delayed effects. Chronic toxicity tests provide data on long-term effects.  
5.3 A life-cycle toxicity test is conducted to determine the effects of the test material on survival, growth, and reproduction of the test species. Additional sublethal endpoints (for example, biochemical, physiological, and histopathological) may be used to determine the health of the species under field conditions.  
5.4 The results of acute, chronic, and life-cycle toxicity tests can be used to predict effects likely to occur on marine organisms under field conditions.  
5.5 The results of acute, chronic, or life-cycle toxicity tests might be used to compare the sensitivities of different species and the toxicities of different test materials, as well as to study the effects of various environmental factors on the results of such tests.  
5.6 The results of acute, chronic, or li...
SCOPE
1.1 This guide covers procedures for obtaining data concerning the adverse effects of a test material added to marine and estuarine waters on certain species of polychaetes during short- or long-term continuous exposure. The polychaete species used in these tests are either field collected or from laboratory cultures and exposed to varying concentrations of a toxicant in static or static-renewal conditions. These procedures may be useful for conducting toxicity tests with other species of polychaetes, although modifications might be necessary.  
1.2 Modifications of these procedures might be justified by 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 results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting acute, chronic, or life-cycle tests with other species of polychaetes.  
1.3 These procedures are applicable to most chemicals, either individually or in formulations, commercial products, and known or unknown mixtures. With appropriate modifications, these procedures can be used to conduct these tests on factors such as temperature, salinity, and dissolved oxygen. These procedures can also be used to assess the toxicity of potentially toxic discharges such as municipal wastes, sediments/soils, oil drilling fluids, produced water from oil well production, and other types of industrial wastes. An LC50 (medial lethal concentration) may be calculated from the data generated in each acute and chronic toxicity test when multiple concentrations are tested. Growth, determined by a change in measured weight, and reproduction, as the change in total number of organisms, are used to measure the effect of a toxicant...

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SIGNIFICANCE AND USE
5.1 This test provides an easy and reliable method for the detection of L. pneumophila in potable and non-potable waters in 7 days.  
5.2 Routine monitoring for L. pneumophila determines whether implemented control measures are effective, such as those outlined in a water safety program (2).  
5.2.1 Water system management is necessary to maintain L. pneumophila concentrations below hazardous levels. Through routine measurement of L. pneumophila levels, a monitoring program can ensure that control measures are effective and implemented when necessary in response to increasing levels. Water samples may be examined for L. pneumophila during epidemiological investigations as part of local authority, industrial, or hospital programs, or in order to validate treatment control methods. Routine sampling could also be carried out based on risk assessments or on local, state, or federal requirements or guidelines.
SCOPE
1.1 This test method describes a simple procedure for the detection of Legionella pneumophila (L. pneumophila) in potable water and non-potable waters (cooling towers, for example). This procedure describes a liquid culture method based on a bacterial enzyme technology. The detection of L. pneumophila is signaled through the utilization of a substrate present in the Legiolert reagent. L. pneumophila cells grow rapidly and reproduce using the rich supply of amino acids, vitamins and other nutrients present in the Legiolert reagent. Actively growing strains of L. pneumophila use the added substrate to produce a brown color indicator or produce turbid growth with or without brown coloration. Legiolert can detect this bacterial species at the following minimum concentrations based on the protocol employed:  
1.1.1 Potable Water:  
1.1.1.1 ≥1 organism / 100 mL at 7 days for 100 mL potable protocol.
1.1.1.2 ≥1 organism / 10 mL at 7 days for 10 mL potable protocol.  
1.1.2 Non-potable Water:  
1.1.2.1 ≥1 organism / 1.0 mL at 7 days for 1.0 mL non-potable protocol.
1.1.2.2 ≥1 organism / 0.1 mL at 7 days for 0.1 mL non-potable protocol.  
1.1.3 This test method can be used for potable (drinking) waters and non-potable waters such as cooling tower waters (1).3 It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 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.  
1.4 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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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...

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SIGNIFICANCE AND USE
5.1 An acute toxicity test is conducted to assess effects of a short-term exposure of organisms to a test material under specific experimental conditions. An acute toxicity test does not provide information concerning whether delayed effects will occur and typically evaluates effects on survival. A chronic test is typically longer in duration and includes a sublethal endpoint to assess effects on a population that might occur beyond the exposure period. Because the echinoderm embryo development test includes a sublethal endpoint, but is also short in duration, these tests are considered to be short-term chronic tests, consistent with EPA guidance.  
5.2 Because embryos and larvae are usually assumed to be the most sensitive life stages of these echinoid species, and because some of these species are commercially and recreationally important, the results of these tests are often considered to be a good indication of the acceptability of pollutant concentrations to saltwater species in general. The results of these toxicity tests are often assumed to be an important consideration when assessing the hazard of materials to other saltwater organisms (see Guides E724 and E1023) or when deriving water quality criteria for saltwater organisms  (7).  
5.3 The results of short-term chronic toxicity tests might be used to predict effects likely to occur to aquatic organisms in field situations as a result of exposure under comparable conditions, except that toxicity to benthic species might depend on sorption or settling of the test material onto the substrate.  
5.4 The results of short-term chronic tests might be used to compare the sensitivities of different species and the acute toxicities of different test materials, and to determine the effects of various environmental factors on the results of such tests.  
5.5 The results of short-term chronic toxicity tests might be useful for studying the biological availability of, and structure-activity relationships between, t...
SCOPE
1.1 This guide covers procedures for obtaining laboratory data concerning the short-term chronic effects of a test material on echinoderm embryos and the resulting larvae (sea urchins and sand dollars) during static 48- to 96-h exposures. These procedures have generally been used with U.S. East Coast (Arbacia punctulata and  Strongylocentrotus droebachiensis ) (1)3 and West Coast species (Strongylocentrotus purpuratus, S. droebachiensis, and Dendraster excentricus) (2). The basic procedures described in this guide first originated in Japan and Scandanavia  (3), and parallel procedures have been used with foreign species, especially in Japan and the Mediterranean  (4). These procedures will probably be useful for conducting static toxicity tests with embryos of other echinoid species, although modifications might be necessary.  
1.2 Other modifications of these procedures might be justified by special needs or circumstances. Although using procedures appropriate to a particular species or special needs and circumstances is more important than following prescribed procedures, the results of tests conducted by using unusual procedures are not likely to be comparable with those of many other tests. The comparison of results obtained by using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting tests starting with embryos of echinoids.  
1.3 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, and pH and on such materials as aqueous effluents (see also Guide E1192), leachates, oils, particulate matter, surface waters, effluents, and sediments (Annex A1). Renewal tests might be preferable to static tests for materials that have a high oxygen demand, are...

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SIGNIFICANCE AND USE
5.1 An acute toxicity test is conducted to assess the effects of a short term exposure of organisms to a test material under specific experimental conditions. An acute toxicity test does not provide information concerning whether delayed effects will occur and typically evaluates effects on survival. A chronic test is typically longer in duration and includes a sublethal endpoint to assess effects on a population that might occur beyond the exposure period. Because the bivalve embryo development test includes a sublethal endpoint, but is also short in duration, these tests are considered to be short-term chronic tests.  
5.2 Because embryos and larvae are usually assumed to be the most sensitive life stages of these bivalve mollusc species and because these species are commercially and recreationally important, results of these acute tests are often considered to be a good indication of the acceptability of pollutant concentrations to saltwater molluscan species in general. Results of these acute toxicity tests are often assumed to be an important consideration when assessing the hazard of materials to other saltwater organisms (see Guide E1023) or when deriving water quality criteria for saltwater organisms (3) .  
5.3 Results of short-term chronic toxicity tests might be used to predict effects likely to occur to aquatic organisms in field situations as a result of exposure under comparable conditions, except that toxicity to benthic species might depend on sorption or settling of the test material onto the substrate.  
5.4 Results of short-term chronic tests might be used to compare the sensitivities of different species to different test materials, and to determine the effects of various environmental factors on results of such tests.  
5.5 Results of short-term chronic toxicity tests might be useful for studying biological availability of, and structure activity relationships between, test materials.  
5.6 Results of any toxicity test will depend on temper...
SCOPE
1.1 This guide describes procedures for obtaining laboratory data concerning the acute effects of a test material on embryos and the resulting larvae of four species of saltwater bivalve molluscs (Pacific oyster, Crassostrea gigas Thunberg; eastern oyster,  Crassostrea virginica Gmelin; quahog or hard clam,  Mercenaria mercenaria Linnaeus; and the mussel species complex (Mytilus spp.) including the blue mussel,  Mytilus edulis Linnaeus; the Mediterranean mussel, Mytilus galloprovincialis Lamark; and the Northern Bay Mussel, Mytilus trossulus Gould) during static 48-h exposures. These procedures will probably be useful for conducting static short-term chronic toxicity tests starting with embryos of other bivalve species (1)2 although modifications might be necessary.  
1.2 Other modifications of these procedures might be justified by special needs or circumstances. Although using procedures appropriate to a particular species or special needs and circumstances is more important than following prescribed procedures, results of tests conducted by using unusual procedures are not likely to be comparable to results of many other tests. Comparison of results obtained by using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting 48-h acute tests starting with embryos of bivalve molluscs.  
1.3 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 acute tests on temperature, dissolved oxygen, and pH and on such materials as aqueous effluents (see also Guide E1192), leachates, oils, particulate matter, sediments, and surface waters. Renewal tests might be preferable to static tests for materials that have a high oxygen demand, are highly volatile, are rapidly biologically or chemically transformed...

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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).  
5.2.4 Life-cycle tests are used to evaluate the biological availability of, and structure-activity relationships between, test materials and t...
SCOPE
1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of a test material (added to dilution water, but not to food) on Daphnia magna Straus, 1820, during continuous exposure throughout a life-cycle using the renewal or flow-through techniques. These procedures also should be useful for conducting life-cycle toxicity tests with other invertebrate species and cladocerans from the same genus (for example, Daphnia pulex), although modifications might be necessary.  
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.  
1.3 Modification 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 standard test procedures. Comparison of results obtained using modified and unmodified versions of these procedures might provide useful information on new concepts and procedures for conducting life-cycle toxicity tests with D. magna. Appendix X3 provides modifications for conducting the chronic toxicity test method with D. pulex Leydig, 1860.  
1.4 This guide is arranged as follows:    
Section  
Referenced Documents  
2  
Terminology  
3  
Summary of Guide  
4  
Si...

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SIGNIFICANCE AND USE
5.1 This test method is designed to assess the retentivity of a sterilizing filter under standard challenge conditions.  
5.1.1 A challenge of 107 bacteria per cm2 of effective filtration area is selected to provide a high degree of assurance that the method has sufficient sensitivity to detect oversized pores and that the filter will quantitatively retain large numbers of organisms. The model challenge organism, B. diminuta, is widely considered to be a small bacterium and is recognized as an industry standard for qualifying sterilizing filters. Other species may represent a worst-case test in terms of ability to penetrate a filter. This test does not provide assurance that filters can completely retain such bacteria.  
5.1.2 The analytical procedure utilized in this test method provides a method to assign a numerical value to the filtration efficiency of the filter being evaluated under standard filtration conditions. For the purpose of product sterility assurance, additional process-specific studies should be performed.
SCOPE
1.1 This test method determines the bacterial retention characteristics of membrane filters for liquid filtration using Brevundimonas diminuta as the challenge organism. This test method can be used to evaluate any membrane filter system used for liquid sterilization.  
1.2 This test method is not intended to be used in performance of product- and process-specific validation of the bacterial retention characteristics of membrane filters to be used in pharmaceutical or biopharmaceutical sterilizing filtration, or both. Process- and product-specific bacterial retention validation should be carried out using the intended product manufacturing process parameters and the product solution or surrogate as the carrier fluid.  
1.3 The values stated in SI units are to be regarded as standard.  
1.3.1 Exception—The inch-pound values given for units of pressure are to be regarded as standard; SI unit conversions are shown in parentheses.  
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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SIGNIFICANCE AND USE
4.1 Numerous ASTM test methods and practices (for example: Test Methods D5259 and D5392, and Practices D6974 and E2563) report colony counts as their measured parameter.  
4.2 These practices provide a uniform set of counting, calculating, and reporting procedures for ASTM test methods in microbiology.    
Section  
A—Counting Colonies on Membrane Filters  
6  
B—Counting Colonies on Pour Plates  
7  
C—Counting Colonies on Spread Plates  
8  
4.3 The counting rules provide a best attainable estimate of microorganisms in the sample, since the samples cannot be held and reanalyzed at a later date.
SCOPE
1.1 These practices cover recommended procedures for counting colonies and reporting colony-forming units (CFU) on membrane filters (MF) and standard pour and spread plates.  
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3 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.  
1.4 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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SIGNIFICANCE AND USE
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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This document specifies a method for the detection, semi-quantitative and quantitative (MPN) enumeration of thermotolerant Campylobacter species.  
The method can be applied to all kinds of waters including: drinking water, ground water and well water, fresh, brackish and saline surface water, swimming pools, spa and hydrotherapy pools, recreational waters, agricultural waters and runoff, untreated and treated wastewater and also sand and other sediments.  
This method can be used for the detection of Campylobacter species in a specified sample volume. Clean water samples with low turbidity can be membrane filtered for either a qualitative method, semi-quantitative or quantitative (MPN) method. Water samples with higher turbidity, such as primary and secondary wastewater effluents and sediments, are analysed using the same qualitative, semi-quantitative or quantitative MPN method by direct inoculation of material into bottles or tubes. Sediments can be suspended in a suitable diluent or inoculated directly into enrichment broths.
Users wishing to employ this method are expected to verify its performance for the particular matrix under their own laboratory conditions.

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This document specifies a method for the detection, semi-quantitative and quantitative (MPN) enumeration of thermotolerant Campylobacter species. The method can be applied to all kinds of waters including: drinking water, ground water and well water, fresh, brackish and saline surface water, swimming pools, spa and hydrotherapy pools, recreational waters, agricultural waters and runoff, untreated and treated wastewater and also sand and other sediments. This method can be used for the detection of Campylobacter species in a specified sample volume. Clean water samples with low turbidity can be membrane filtered for either a qualitative method, semi-quantitative or quantitative (MPN) method. Water samples with higher turbidity, such as primary and secondary wastewater effluents and sediments, are analysed using the same qualitative, semi-quantitative or quantitative MPN method by direct inoculation of material into bottles or tubes. Sediments can be suspended in a suitable diluent or inoculated directly into enrichment broths. Users wishing to employ this method are expected to verify its performance for the particular matrix under their own laboratory conditions.

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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.  
5.1.1 This procedure as devised will enable the user to test each membrane filter lot number for its ability to retain all bacterial equal to, or larger than, the stated membrane pore size.  
5.2 Since this membrane is often used to sterilize nonautoclavable liquids, it is essential that the retention characteristics of this membrane are stable.
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 the 0.2-µm pore size of the membrane filter.  
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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This document specifies a method for the detection and quantification of Legionella spp. and L. pneumophila using a quantitative polymerase chain reaction (qPCR). It specifies general methodological requirements, performance evaluation requirements, and quality control requirements.
Technical details specified in this document are given for information only. Any other technical solutions complying with the performance requirements are suitable.
NOTE 1 For performance requirements, see Clause 9.
This document is intended to be applied in the bacteriological investigation of all types of water (hot or cold water, cooling tower water, etc.), unless the nature and/or content of suspended matter and/or accompanying flora interfere with the determination. This interference can result in an adverse effect on both the detection limit and the quantification limit.
NOTE 2 For validation requirements, see 9.7.
The results are expressed as the number of genome units of Legionella spp. and/or L. pneumophila per litre of sample.
The method described in this document is applicable to all types of water. However, some additives, such as chemicals used for water treatment, can interfere with and/or affect the sensitivity of the method.
The qPCR methods do not give any information about the physiological state of the Legionella.

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This document specifies requirements and gives guidance for performing the manipulations common to each culture technique for the microbiological examination of water, particularly the preparation of samples, culture media, and general apparatus and glassware, unless otherwise required in the specific standard. It also describes the various techniques available for detection and enumeration by culture and the criteria for determining which technique is appropriate.
This document is mainly intended for examinations for bacteria, yeasts and moulds, but some aspects are also applicable to bacteriophages, viruses and parasites. It excludes techniques not based on culturing microorganisms, such as polymerase chain reaction (PCR) methods.

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This document specifies requirements and gives guidance for performing the manipulations common to each culture technique for the microbiological examination of water, particularly the preparation of samples, culture media, and general apparatus and glassware, unless otherwise required in the specific standard. It also describes the various techniques available for detection and enumeration by culture and the criteria for determining which technique is appropriate.
This document is mainly intended for examinations for bacteria, yeasts and moulds, but some aspects are also applicable to bacteriophages, viruses and parasites. It excludes techniques not based on culturing microorganisms, such as polymerase chain reaction (PCR) methods.

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This document specifies a method for the detection and quantification of Legionella spp. and L. pneumophila using a quantitative polymerase chain reaction (qPCR). It specifies general methodological requirements, performance evaluation requirements, and quality control requirements. Technical details specified in this document are given for information only. Any other technical solutions complying with the performance requirements are suitable. NOTE 1 For performance requirements, see Clause 9. This document is intended to be applied in the bacteriological investigation of all types of water (hot or cold water, cooling tower water, etc.), unless the nature and/or content of suspended matter and/or accompanying flora interfere with the determination. This interference can result in an adverse effect on both the detection limit and the quantification limit. NOTE 2 For validation requirements, see 9.7. The results are expressed as the number of genome units of Legionella spp. and/or L. pneumophila per litre of sample. The method described in this document is applicable to all types of water. However, some additives, such as chemicals used for water treatment, can interfere with and/or affect the sensitivity of the method. The qPCR methods do not give any information about the physiological state of the Legionella.

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  • Technical specification
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SIGNIFICANCE AND USE
5.1 Sulfate reducing archaea and bacteria are known to contribute to microbiologically influenced corrosion.  
5.2 Sulfate-reducing bacteria are widely distributed in marine and fresh water muds which, in consequence, frequently are laden with the hydrogen sulfide produced by these organisms during dissimilatory sulfate reduction.  
5.3 Traditional, culture-dependent methods such as those described in Test Methods D4412, prescribe incubation periods of as long as 21 days before assigning a below detection limit (BDL) score to a specimen. Moreover, it is well known that not all SRP will proliferate in the nutrient media specified in Test Methods D4412.  
5.4 This test method uses ELISA technology to provide semi-quantitative, culture-independent, SRP bioburden test results in less than 30 min.  
5.4.1 Because all the reagents and supplies used are non-hazardous and prepackaged for single test use, this test method does not require any apparatus other than a laboratory timer. Consequently, it can be performed at or near the point of sample collection.  
5.4.2 The opportunity to minimize the delay between sample collection, testing, and results availability translates into timely use of the data to drive preventive and corrective SRB control measures.
SCOPE
1.1 This test method provides a protocol for using enzyme-linked immunosorbent assay (ELISA) technology to test water samples for the enzyme adenosine 5’-phosphosulfate reductase (APSr) concentration.  
1.1.1 APSr is present in all known sulfate reducing protists (SRP – sulfate reducing bacteria – SRB – and sulfate reducing archaea – SRA).  
1.1.2 As reported in U.S. Patent 4,999,286, APS reductase concentration can be used as a surrogate parameter for estimating SRA bioburdens (Appendix X1 compares results from Test Methods D8243, D4412, and quantitative polymerase chain reaction – qPCR – testing).  
1.2 This test method has been validated in tap water, oilfield produced water (salinities ranging from 100 g L-1 to 600 g L-1), and fuel-associated water (commonly referred to as bottoms-water).  
1.3 This test method detects APS reductase semi-quantitatively in the range of 0.001M to 0.1M – correlating to 102 SRP/mL to 106 SRP/mL.  
1.3.1 As described in Appendix X2 test method sensitivity can be increased 10-fold to 100-fold. However, the precision statistics provided in X apply only to 10-mL specimens.  
1.4 The values stated in SI units are to be regarded as 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. Some specific hazards statements are given in Section 9 on Hazards.  
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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This document specifies requirements and gives guidance for performing the manipulations common to each culture technique for the microbiological examination of water, particularly the preparation of samples, culture media, and general apparatus and glassware, unless otherwise required in the specific standard. It also describes the various techniques available for detection and enumeration by culture and the criteria for determining which technique is appropriate. This document is mainly intended for examinations for bacteria, yeasts and moulds, but some aspects are also applicable to bacteriophages, viruses and parasites. It excludes techniques not based on culturing microorganisms, such as polymerase chain reaction (PCR) methods.

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ISO 13843:2017 deals with characterization of microbiological methods. In terms of ISO 13843:2017, characterization means the study of parameters that can be measured to describe how the method is likely to perform in a given set of conditions, which can be described as performance characteristics. ISO 13843:2017 describes procedures for the determination of performance characteristics which can be used for subsequent validation or verification of methods.
The emphasis is on selective quantitative methods and ISO 13843:2017 applies to all types of water. For methods that are not based upon direct microscopic count, colony count or most probable number, the applicability of the procedures described in ISO 13843:2017 should be considered carefully.

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ISO 11731:2017 specifies culture methods for the isolation of Legionella and estimation of their numbers in water samples.
These methods are applicable to all kinds of water samples including potable, industrial, waste and natural waters. These methods can be used for water related matrices, e.g. biofilms, sediments, etc.
Not all Legionella species are culturable; therefore, the methods described in this document do not recover all species of Legionella.

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ISO 20227:2017 specifies a method for the determination of the inhibition of the growth of the first fronds of Spirodela polyrhiza germinated from turions, by substances and mixtures contained in water or waste water, including treated municipal waste water and industrial effluents.
The test is also applicable to pure chemicals and in particular, plant protection products and pesticides.

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ISO 13843:2017 deals with characterization of microbiological methods. In terms of ISO 13843:2017, characterization means the study of parameters that can be measured to describe how the method is likely to perform in a given set of conditions, which can be described as performance characteristics. ISO 13843:2017 describes procedures for the determination of performance characteristics which can be used for subsequent validation or verification of methods.
The emphasis is on selective quantitative methods and ISO 13843:2017 applies to all types of water. For methods that are not based upon direct microscopic count, colony count or most probable number, the applicability of the procedures described in ISO 13843:2017 should be considered carefully.

  • Standard
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ISO 11731:2017 specifies culture methods for the isolation of Legionella and estimation of their numbers in water samples.
These methods are applicable to all kinds of water samples including potable, industrial, waste and natural waters. These methods can be used for water related matrices, e.g. biofilms, sediments, etc.
Not all Legionella species are culturable; therefore, the methods described in this document do not recover all species of Legionella.

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