This document is focused on the structural features of rivers, on geomorphological and hydrological processes, and on river continuity. It provides guidance on the features and processes to be taken into account when characterizing and assessing the hydromorphology of rivers. It is based on methods developed, tested, and compared in Europe. Its main aim is to improve the comparability of hydromorphological assessment methods, data processing and interpretation. Although it has particular importance for the WFD by providing guidance on assessing hydromorphological quality, it has considerably wider scope for other applications. In addition, while recognizing the important influence of hydromorphology on plant and animal ecology, no attempt is made to provide guidance in this area, but where the biota have an important influence on hydromorphology these influences are included.
NOTE   A case study illustrating the application of this standard is given in Gurnell and Grabowski[1].

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This document specifies a method for determining the toxicity of environmental samples on growth, fertility and reproduction of Caenorhabditis elegans. The method applies to contaminated whole freshwater sediment (maximum salinity 5 g/l), soil and waste, as well as to pore water, elutriates and aqueous extracts that were obtained from contaminated sediment, soil and waste.

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This standard 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, semiquantitative or quantitative (MPN) method. Water samples with higher turbidity, such as primary and secondary wastewater effluents and sediments, are analysed using the same qualitative, semiquantitative 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 provides guidance for survey design, equipment specification, survey methods, sampling and data handling of macroalgae and marine angiosperms such as Zostera in the intertidal soft bottom environment. It does not include polyeuryhaline terrestrial angiosperms that are found in saltmarshes. Ruppia is a genus of angiosperms that can be found in brackish water. This document can also be applied to the study of Ruppia in these environments.
The document comprises:
-   development of a mapping and sampling programme;
-   requirements for mapping and sampling equipment;
-   procedures for remote sensing data collection;
-   procedures for direct mapping and sampling in the field;
-   recommendations for taxon identification and biomass determination;
-   data handling.

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This document provides guidance for survey design, equipment specification, survey methods, sampling and data handling of macroalgae and marine angiosperms such as Zostera in the intertidal soft bottom environment. It does not include polyeuryhaline terrestrial angiosperms that are found in saltmarshes. Ruppia is a genus of angiosperms that can be found in brackish water. This document can also be applied to the study of Ruppia in these environments.
The document comprises:
-   development of a mapping and sampling programme;
-   requirements for mapping and sampling equipment;
-   procedures for remote sensing data collection;
-   procedures for direct mapping and sampling in the field;
-   recommendations for taxon identification and biomass determination;
-   data handling.

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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 technical report specifies the data and metadata necessary to validate the identity of a diatom barcode along with recommendations for storage of the barcode and metadata to ensure access to this information.

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This technical report specifies a method for the field sampling of benthic diatoms which will be then analysed by subsequent metabarcoding techniques for ecological status and water quality assessments. Data produced by this method are suitable for production of taxonomical diatom lists.

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This European standard specifies a procedure for analysing mesozooplankton in marine and brackish waters. The procedures comprise how to identify and enumerate zooplankton to estimate quantitative information on diversity, abundance and biomass with regard to spatial distribution and long-term temporal trends for a given body of water.

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This document specifies a method for the sampling of mesozooplankton from marine and brackish waters using mesh.

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This document specifies procedures for sampling of mesozooplankton using nets and continuous ribbon-sampling devices in marine and brackish waters for the purpose of water quality assessment and determination of ecological status of ecosystems.
Guidance on sampling procedures and the subsequent steps of preservation and storage are given. The sampling procedures allow estimates of species occurrence and their abundance (relative or absolute), including spatial distribution and seasonal and long-term temporal trends, for a given body of water.
The described methods are restricted to the sampling of mesozooplankton that inhabit marine and brackish waters and exclude the shallow littoral zones which require a different type of sampling (e.g. zooplankton in salt marshes).

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This document specifies a method for the enumeration of Pseudomonas aeruginosa in water. The
method is based on the growth of target organisms in a liquid medium and calculation of the most
probable number (MPN) of organisms by reference to MPN tables.
This document is applicable to a range of types of water. For example, hospital waters, drinking water
and non‐carbonated bottled waters intended for human consumption, groundwater, swimming pool
and spa pool waters including those containing high background counts of heterotrophic bacteria.
This document does not apply to carbonated bottled waters, flavoured bottle waters, cooling tower
waters or marine waters, for which the method has not been validated. These waters are, therefore,
outside the scope of this document. Laboratories can employ the method presented in this document
for these matrices by undertaking appropriate validation of performance of this method prior to use.
The test is based on a bacterial enzyme detection technology that signals the presence of P. aeruginosa
through the hydrolysis of a 7‐amino‐4‐methylcoumarin aminopeptidase substrate present in a special
reagent. P. aeruginosa cells rapidly grow and reproduce using the rich supply of amino acids, vitamins
and other nutrients present in the reagent. Actively growing strains of P. aeruginosa have an enzyme
that cleaves the 7‐amido‐coumarin aminopeptidase substrate releasing a product which fluoresces
under ultraviolet (UV) light. The test described in this document provides a confirmed result within
24 h with no requirement for further confirmation of positive wells.

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This standard gives guidance on the estimation of abundance and identification of macro-invertebrates in samples taken from inland waters. The procedure deals with pre-treatment (cleaning), sub-sampling, sorting and final identification of organisms from preserved and live samples originating from natural habitats or artificial substrates.

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This document specifies techniques for preparing poorly water-soluble organic compounds (i.e. liquid
and solid compounds) with a solubility in water of less than approximately 100 mg/l and introducing
them into test vessels for a subsequent biodegradability test in an aqueous medium using standard
methods.
The subsequent tests on biodegradability are primarily methods using the analysis of the released
carbon dioxide described in ISO 9439 and the determination of the oxygen described in ISO 9408 and
following the usual precautions for ISO 10707. Thus, one can notice that the methods measuring the
removal of dissolved organic carbon (DOC) are not appropriate.
This document does not specify the biodegradation test methods. It is restricted to describing
techniques for introducing the test compounds into the test medium and to keeping them in a dispersed
state[4]. These techniques are implemented while observing the experimental conditions described
in the standardized methods for evaluating biodegradability. ISO 9439, based on CO2 evolution, is not
suitable for testing volatile compounds.
Some of the preparation methods described in this document might not be accepted by regulators for
making conclusions on the ready biodegradability of tested compounds.
Examples of biodegradability curves are given in Annex A.
2 Normative

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ISO 11348 describes three methods for determining the inhibition of the luminescence emitted by the marine bacterium Vibrio fischeri (NRRL B-11177). This part of ISO 11348 specifies a method using freshly prepared bacteria. The method is applicable to waste water, aqueous extracts and leachates, fresh water (surface and ground water), sea and brackish water, eluates of sediment (fresh water, brackish and sea water), pore water, singles substances, diluted in water.

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This document specifies a procedure for analysing mesozooplankton in marine and brackish waters. The procedure comprises how to identify and enumerate mesozooplankton to estimate quantitative information on diversity, abundance and biomass with regard to spatial distribution and long-term temporal trends for a given body of water.

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This document specifies a method for the determination of fish acute toxicity using the permanent cell line from rainbow trout (Oncorhynchus mykiss) gill, RTgill-W1. Cells in confluent monolayers in 24-well tissue culture plates are exposed to water samples, such as surface waters or different kinds of effluents, or to chemicals for 24 h and, thereafter, cell viability is assessed based on fluorescent cell viability indicator dyes (see 4.1). Data are then expressed as a percentage of unexposed control and toxicity quantified based on the percentage of cell viability versus the percentage of effluent or the chemical concentration in response curves (see Clause 9).

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This document specifies techniques for preparing poorly water-soluble organic compounds (i.e. liquid and solid compounds) with a solubility in water of less than approximately 100 mg/l and introducing them into test vessels for a subsequent biodegradability test in an aqueous medium using standard methods.
The subsequent tests on biodegradability are primarily methods using the analysis of the released carbon dioxide described in ISO 9439 and the determination of the oxygen described in ISO 9408 and following the usual precautions for ISO 10707. Thus, one can notice that the methods measuring the removal of dissolved organic carbon (DOC) are not appropriate.
This document does not specify the biodegradation test methods. It is restricted to describing techniques for introducing the test compounds into the test medium and to keeping them in a dispersed state[4]. These techniques are implemented while observing the experimental conditions described in the standardized methods for evaluating biodegradability. ISO 9439, based on CO2 evolution, is not suitable for testing volatile compounds.
Some of the preparation methods described in this document might not be accepted by regulators for making conclusions on the ready biodegradability of tested compounds.
Examples of biodegradability curves are given in Annex A.

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This document specifies techniques for preparing poorly water-soluble organic compounds (i.e. liquid and solid compounds) with a solubility in water of less than approximately 100 mg/l and introducing them into test vessels for a subsequent biodegradability test in an aqueous medium using standard methods. The subsequent tests on biodegradability are primarily methods using the analysis of the released carbon dioxide described in ISO 9439 and the determination of the oxygen described in ISO 9408 and following the usual precautions for ISO 10707. Thus, one can notice that the methods measuring the removal of dissolved organic carbon (DOC) are not appropriate. This document does not specify the biodegradation test methods. It is restricted to describing techniques for introducing the test compounds into the test medium and to keeping them in a dispersed state[4]. These techniques are implemented while observing the experimental conditions described in the standardized methods for evaluating biodegradability. ISO 9439, based on CO2 evolution, is not suitable for testing volatile compounds. Some of the preparation methods described in this document might not be accepted by regulators for making conclusions on the ready biodegradability of tested compounds. Examples of biodegradability curves are given in Annex A.

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This technical report specifies the data and metadata necessary to validate the identity of a diatom barcode used for ecological assessment along with recommendations for storage of the barcode and metadata to ensure access to this information.

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This technical report specifies a method for the field sampling of benthic diatoms which will be then analysed by subsequent metabarcoding techniques for ecological status and water quality assessments. Data produced by this method are suitable for production of taxonomical diatom lists.

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This document specifies a method for the determination of the estrogenic potential of water and waste water by means of a reporter gene assay utilizing stably transfected human cells. This reporter gene assay is based on the activation of the human estrogen receptor alpha. This method is applicable to: — fresh water; — waste water; — aqueous extracts and leachates; — eluates of sediments (fresh water); — pore water; — aqueous solutions of single substances or of chemical mixtures; — drinking water; — the limit of quantification (LOQ) of this method for the direct analysis of water samples is between 0,3 ng/l and 1 ng/l 17β-estradiol equivalents (EEQ) based on the results of the international interlaboratory trial (see Annex F). The upper working range was evaluated [based on the results of the international interlaboratory trial (see Table F.3)] up to a level of 75 ng EEQ/l. Samples showing estrogenic potencies above this threshold have to be diluted for a valid quantification. Extraction and pre concentration of water samples can prove necessary if their estrogenic potential is below the given LOQ.

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This document specifies a method for the determination of the estrogenic potential of water and waste water by means of a reporter gene assay with a genetically modified yeast strain Arxula adeninivorans. This reporter gene assay is based on the activation of the human estrogen receptor alpha. Arxula adeninivorans is a highly robust and salt- and temperature-tolerant test organism and is especially suitable for the analysis of samples with high salinity (conductivity up to 70 mS/cm). The test organism can be cultivated in medium with sodium chloride content up to 20 %. This method is applicable to: — fresh water; — waste water; — sea water; — brackish water; — aqueous extracts and leachates; — eluates of sediments (fresh water); — pore water; — aqueous solutions of single substances or of chemical mixtures; — drinking water. The limit of quantification (LOQ) of this method for the direct analysis of water samples is between 1,5 ng/l and 3 ng/l 17β-estradiol equivalents (EEQ). The upper threshold of the dynamic range for this test is between 25 ng/l and 40 ng/l 17β-estradiol equivalents (EEQ). Samples showing estrogenic potencies above this threshold have to be diluted for a valid quantification. Extraction and pre-concentration of water samples can prove necessary, if their estrogenic potential is below the given LOQ. An international interlaboratory trial for the validation of this document has been carried out. The results are summarized in Annex F. NOTE Extraction and pre-concentration of water samples can prove necessary.

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This document specifies a method for the determination of the estrogenic potential of water and waste water by means of a reporter gene assay with genetically modified yeast strains Saccharomyces cerevisiae. This reporter gene assay is based on the activation of the human estrogen receptor alpha. This method is applicable to: — fresh water; — waste water; — aqueous extracts and leachates; — eluates of sediments (fresh water); — pore water; — aqueous solutions of single substances or of chemical mixtures; — drinking water. The limit of quantification (LOQ) of this method for the direct analysis of water samples is between 8 ng/l and 15 ng/l 17β-estradiol equivalents (EEQ) based on the results of the international interlaboratory trial (see Annex F). The upper threshold of the dynamic range for this test is between 120 ng/l and 160 ng/l 17β-estradiol equivalents (EEQ). Samples showing estrogenic potencies above this threshold have to be diluted for a valid quantification. Extraction and pre-concentration of water samples can prove necessary, if their estrogenic potential is below the given LOQ.

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This document specifies a method for the enumeration of Pseudomonas aeruginosa in water. The method is based on the growth of target organisms in a liquid medium and calculation of the most probable number (MPN) of organisms by reference to MPN tables. This document is applicable to a range of types of water. For example, hospital waters, drinking water and non‑carbonated bottled waters intended for human consumption, groundwater, swimming pool and spa pool waters including those containing high background counts of heterotrophic bacteria. This document does not apply to carbonated bottled waters, flavoured bottle waters, cooling tower waters or marine waters, for which the method has not been validated. These waters are, therefore, outside the scope of this document. Laboratories can employ the method presented in this document for these matrices by undertaking appropriate validation of performance of this method prior to use. The test is based on a bacterial enzyme detection technology that signals the presence of P. aeruginosa through the hydrolysis of a 7‑amino‑4‑methylcoumarin aminopeptidase substrate present in a special reagent. P. aeruginosa cells rapidly grow and reproduce using the rich supply of amino acids, vitamins and other nutrients present in the reagent. Actively growing strains of P. aeruginosa have an enzyme that cleaves the 7‑amido‑coumarin aminopeptidase substrate releasing a product which fluoresces under ultraviolet (UV) light. The test described in this document provides a confirmed result within 24 h with no requirement for further confirmation of positive wells.

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This International Standard specifies a method for the determination of the inhibition of the growth of the first fronds of Spirodela polyrhiza germinated from dormant 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 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 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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This document gives practical guidance on sampling, pre-treatment, performance and evaluation of
environmental samples in the context of performing biological tests. Information is given on how to
cope with the problems of biotesting arising from the sample and the suitability of the test design.
It is intended to convey practical experience concerning precautions to be taken by describing methods
successfully proven to solve or to circumvent some of the experimental problems of biotesting of, for
example, waters.
Primarily dealt with are substance-related problems concerning sampling and pre-treatment of
environmental samples (e.g. waste water samples) for the performance of biotests.
This guidance is on ecotoxicological testing with organisms (single-species biotests; in vivo and in
vitro). Some features addressed in this document also apply to biotests using single-cell systems (in
vitro bioassays) and biodegradation studies as far as sampling and sample preparations are concerned.
Testing of substances in the water solubility range is also addressed.
Reference has been made as far as possible to existing International Standards and guidelines.
Information taken from published papers or oral communication has been utilized as well.
This document is applicable to biological tests for determining the effect of environmental samples
like treated communal and industrial waste water, groundwater, fresh water, aqueous extracts (e.g.
leachates, eluates), pore water of sediments and whole sediments. This document is also applicable to
chemical substances.
This document is not applicable to bacteriological examination of water. Appropriate methods for
bacteriological examination are described in other documents (see ISO 19458[17]).

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This document specifies a method for the determination of the inhibition of growth of the unicellular
marine algae Skeletonema sp. and Phaeodactylum tricornutum by substances and mixtures contained in
sea water or by environmental water samples (effluents, elutriates, etc.).
The method can be used for testing substances that are readily soluble in water and are not significantly
degraded or eliminated in any other way from the test medium.
NOTE With modifications, as described in ISO 14442 and ISO 5667–16, the inhibitory effects of poorly
soluble organic and inorganic materials, volatile compounds, metal compounds, effluents, marine water samples
and elutriates of sediments can be tested.

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This standard provides guidance on methods for monitoring freshwater pearl mussel populations and the environmental characteristics important for maintaining populations in favourable condition. The standard is based on best practice developed and used by Margaritifera experts in Europe, and describes approaches that individual countries have adopted for survey, data analysis and condition assessment. While it is recommended that the causes for pearl mussel decline should be urgently investigated, standard methods for restoring populations are beyond the scope of this document.

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ISO 5667-16:2017 gives practical guidance on sampling, pre-treatment, performance and evaluation of environmental samples in the context of performing biological tests. Information is given on how to cope with the problems of biotesting arising from the sample and the suitability of the test design.
It is intended to convey practical experience concerning precautions to be taken by describing methods successfully proven to solve or to circumvent some of the experimental problems of biotesting of, for example, waters.
Primarily dealt with are substance-related problems concerning sampling and pre-treatment of environmental samples (e.g. waste water samples) for the performance of biotests.
This guidance is on ecotoxicological testing with organisms (single-species biotests; in vivo and in vitro). Some features addressed in this document also apply to biotests using single-cell systems (in vitro bioassays) and biodegradation studies as far as sampling and sample preparations are concerned. Testing of substances in the water solubility range is also addressed.
Reference has been made as far as possible to existing International Standards and guidelines. Information taken from published papers or oral communication has been utilized as well.
ISO 5667-16:2017 is applicable to biological tests for determining the effect of environmental samples like treated communal and industrial waste water, groundwater, fresh water, aqueous extracts (e.g. leachates, eluates), pore water of sediments and whole sediments. This document is also applicable to chemical substances.
ISO 5667-16:2017 is not applicable to bacteriological examination of water. Appropriate methods for bacteriological examination are described in other documents (see ISO 19458[17]).

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This European Standard provides guidance on methods for monitoring freshwater pearl mussel populations and the environmental characteristics important for maintaining populations in favourable condition. The standard is based on best practice developed and used by Margaritifera experts in Europe, and describes approaches that individual countries have adopted for survey, data analysis and condition assessment. While it is recommended that the causes for pearl mussel decline should be urgently investigated, standard methods for restoring populations are beyond the scope of this document.

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ISO 10253:2016 specifies a method for the determination of the inhibition of growth of the unicellular marine algae Skeletonema sp. and Phaeodactylum tricornutum by substances and mixtures contained in sea water or by environmental water samples (effluents, elutriates, etc.).
The method can be used for testing substances that are readily soluble in water and are not significantly degraded or eliminated in any other way from the test medium.

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This document provides guidance on methods for sampling invertebrates in the hyporheic zone of wadable rivers. It describes each method, including details of the equipment involved and its use in the field. Guidance is given on developing a sampling strategy and selecting an appropriate survey technique for the purpose of investigation. Benthic macroinvertebrate sampling is covered elsewhere by other published standards (see bibliography). Selected literature with references in support of this document is given in the bibliography.

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ISO 10253:2016 specifies a method for the determination of the inhibition of growth of the unicellular marine algae Skeletonema sp. and Phaeodactylum tricornutum by substances and mixtures contained in sea water or by environmental water samples (effluents, elutriates, etc.). The method can be used for testing substances that are readily soluble in water and are not significantly degraded or eliminated in any other way from the test medium.

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ISO/TS 18220:2016 specifies an early-life stage procedure for determination of the toxic effects of chemicals and water samples on a cold-water brackish water copepod species under semi-static conditions. The biological test variables include survival and development of the early-life stages. The exposure starts with newly hatched ( The benthic living Nitocra complements the planktonic Acartia species in ISO 16778. These organisms represent different life-history strategies as Nitocra is egg-carrying, whereas Acartia is a broadcasting calanoid copepod and thus, different sensitivities of specific life stages. Nitocra is a fresh to brackish water species, which allows testing low salinity waters and is complementary to A. tonsa, which is of marine origin and poorly tolerates low salinities.

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This European Standard provides guidance on methods for sampling invertebrates in the hyporheic zone of wadable rivers. It describes each method, including details of the equipment involved and its use in the field. Guidance is given on developing a sampling strategy and selecting an appropriate survey technique for the purpose of investigation.
NOTE   Benthic macroinvertebrate sampling is covered by other published standards (see Bibliography). Selected literature with references in support of this document is given in the Bibliography.

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ISO 19820:2016 specifies a method for the determination of the lethal effects of toxicants to Brachionus plicatilis after 24 h or 48 h exposure. The method is applicable to the following: a) chemical substances which are soluble or which can be maintained as a stable suspension of dispersions under the conditions of the test; b) industrial or sewage effluents, treated or untreated, if appropriate after decantation, filtration, or centrifugation; c) marine or estuarine waters; d) sediment elutriates/eluates.

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ISO 19827:2016(E) specifies a method for the determination of the lethal effects of toxicants to Brachionus calyciflorus after 24 h exposure. The method is applicable to: a) chemical substances which are soluble, or which can be maintained as a stable suspension of dispersions under the conditions of the test; b) industrial or sewage effluents, treated or untreated, if appropriate after decantation, filtration or centrifugation; c) freshwaters; d) aqueous extracts.

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