CEN/TC 230/WG 2 - Biological methods

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.

This European Standard provides guidance on determining the degree of modification of lake hydromorphological features described in EN 16039. It enables consistent comparisons of hydromorphology between lakes within a country and between different countries in Europe, providing a method for broad based characterization across a wide spectrum of hydromorphological modification. Its primary aim is to assess ‘departure from naturalness’ for a given type of lake as a result of human pressures, and it suggests suitable sources of information that may contribute to characterizing the degree of modification of hydromorphological features. For wholly artificial lakes or reservoirs formed by damming rivers the aim is to assess the extent to which processes approximate to those in comparable natural water bodies. However, this standard does not replace methods that have been developed within particular countries for local assessment and reporting. Decisions on management for individual lakes require expert local knowledge and vary according to lake type.

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.

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.

This European Standard specifies a standardised method for sampling fish in lakes, using benthic multi-mesh gillnets. The method provides a whole-lake estimate for species occurrence, quantitative relative fish abundance and biomass expressed as Catch Per Unit Effort (CPUE), and size structure of fish assemblages in temperate lakes. It also provides estimates, which are comparable over time within a lake and between lakes. This European Standard specifies routines for sampling, data handling and reporting, and provides information on applications and further treatment of data. This European Standard also provides guidance on sampling of fish with pelagic multi-mesh gillnets and sampling of fish for age and growth analyses. Selected references in support of this European Standard are given in the Bibliography.

This European Standard gives guidelines for characterizing the hydromorphology of transitional or coastal (TraC) waters, but does not prescribe detailed methods of assessment. The main aim of this document is to improve the comparability of hydromorphological survey methods, data processing, and the interpretation and presentation of results.
This European Standard:
a)   lists essential features and processes of TraC waters that should be characterized as part of a hydromorphological survey and used for determining hydromorphological condition;
b)   gives guidance on strategies for collecting and presenting hydromorphological data depending on the resources available and the anticipated use of the assessment;
c)   describes how to generate data sets appropriate for monitoring and reporting on the condition of Natura 2000 sites designated under the Habitats Directive and the Birds Directive;
d)   provides guidance on data quality assurance.
This European Standard does not deal with biological assessments in TraC waters such as the presence or absence of individual species or community composition, nor does it attempt to link specific hydromorphological features with their associated biological communities. However, it is relevant where plants or other organisms form significant structural elements of the habitat (e.g. saltmarshes, biogenic reefs).

This European Standard describes the most relevant rules of the Botanical and Zoological Codes necessary for unequivocal recording of biodiversity in the aquatic environment. Furthermore, guidance is given on how to deal with taxonomic changes in relation to recorded taxonomic names.
NOTE   A Code only affects taxonomic changes carried out in the period covered by that particular edition of the Code.

This European Standard specifies a method for the sampling and laboratory preparation of benthic diatoms for ecological status and water quality assessments. Data produced by this method are suitable for production of water quality indices based on the relative abundance of taxa.

ISO 16665:2014 provides guidelines on the quantitative collection and processing of subtidal soft-bottom macrofaunal samples in marine waters.
ISO 16665:2014 encompasses: a) development of the sampling programme; b) requirements for sampling equipment; c) sampling and sample treatment in the field; d) sorting and species identification; e) storage of collected and processed material.
ISO 16665:2014 does not specifically address the following, although some elements may be applicable: bioassay sub-sampling; deep water (>750 m) or offshore sampling; in situ faunal studies, e.g. recolonization assays; non-benthic organisms caught in the sampling device; estuarine sampling; intertidal sampling; meiofaunal sampling and analysis; sampling by dredge and sledge; self-contained underwater breathing apparatus (SCUBA) sampling; statistical design.
Accuracy of position fixing is determined by the geographical area, equipment used and survey objective.

This European Standard specifies methods for the identification and enumeration of relative proportions of diatom taxa on prepared slides and of data interpretation relevant to assessments of water quality in rivers and lakes. It is suitable for use with indices and assessment methods based on the relative abundance of taxa. The methods for identification and enumeration may also be applied to the study of benthic diatoms in other habitats provided that data interpretation methods appropriate to these habitats are used.

This European Standard specifies a method for surveying aquatic macrophytes in running waters for the purpose of assessing ecological status, using these organisms as elements of biological quality. The information provided by this method includes the composition and abundance of the aquatic macrophyte flora.
This European Standard is applicable to all kinds of surface running water bodies, like natural brooks, streams and rivers and their heavily modified equivalents, as well as to artificial water bodies like canals or run-of-river reservoirs.
The general principles of the approach described in this European Standard may also be applied when monitoring water bodies in the fluvial corridor of a river, such as side channels and oxbows.
It is recognized that for a complete assessment of ecological status, other elements of biological quality should also be assessed.

This European Standard defines standard principles for the design of taxonomic keys to ensure proper use of nomenclatural rules and reproducible and traceable identification. These principles also allow for the selection of the best key available.

This European Standard describes methods, requirements and equipment for remote visual surveillance of organisms and the seabed using still photography and video recording to ensure precise and reproducible data. The main aims of the methods are to record or monitor seabed conditions and organisms on and just above the seabed in a reproducible way at a resolution that is appropriate to the aims of the survey.
In caves and overhangs this standard may not be suitable due to technological limitations related to navigation and movement of the observation platform.

This European Standard provides guidance on interlaboratory comparison with a special focus on biological methods. Guidance on the methods and procedures given in this standard should ensure that field survey results and laboratory analyses are comparable within specified limits. This guidance enables participants in interlaboratory comparison to demonstrate their level of performance. In addition it provides a mechanism for quality improvement. This standard describes a process structure. Detailed elements can be found in EN 14996, EN ISO/IEC 17000, EN ISO/IEC 17025, and EN ISO/IEC 17043.

This European Standard provides guidance in the use of in vivo absorption techniques to quantify
chlorophyll-a concentration in marine and fresh waters.
This European Standard is comprised of the following:
-   definition of the equipment requirement;
-   a priori data and mathematical tools;
-   recommendations for verification of measurement system performance and consideration of factors that can influence measurements;
-   listing of the procedures to be implemented.

This European Standard gives guidance on procedures for the pro rata Multi-Habitat-Sampling (MHS) of benthic macro invertebrates in wadeable rivers and streams. The term "pro-rata" reflects the intention to sample adequate proportions of riverine habitats with reference to their percentage occurrence.
The pro-rata MHS technique does not replace other techniques, but is rather, alongside other applications, a fundamental requisite of some multi-metric assessment approaches used to evaluate the ecological status of running waters. The method described in this document is one of the possible techniques among the existing pro-rata MHS techniques.
The MHS methodology is based on Rapid Bioassessment Protocols [1], the procedures of the Environment Agency for England and Wales [2], the Austrian Guidelines for the Assessment of the Saprobiological Water Quality of Rivers and Streams [3], the AQEM sampling manual [4], the AQEM & STAR site protocol [5], EN 27828, the Austrian Standards M 6232 and M 6119-2 [6], [7], the German Standard DIN 38410-1 [8] and the French Standard XP T90-333 [9].
This European Standard also describes in a detailed manner how to sample different habitats that might be suitable for sampling approaches other than Multi-Habitat-Sampling.

This European Standard is applicable to lakes, which are water bodies occupying one or more basins with surface areas greater than 1 ha (0,01 km2) and maximum depths (at mean water level) greater than 1 m. All types of permanent lakes, including natural, modified and artificial, fresh water and brackish, except for those systems which regularly connect to the sea, are included in this European Standard, though canals are excluded.
Based on these criteria, it can be estimated that there are at least 500 000 natural lakes across Europe, most of which are located in the glaciated landscapes in northern and western provinces and in Scandinavia. Lakeland districts also occur locally in areas such as the Danubian plain and around the Alps. Elsewhere, naturally occurring lakes are relatively sparse and in such areas reservoirs or pits are more common.
This European Standard is designed to:
a)   support environmental and conservation agencies in meeting the monitoring requirements of the WFD (Article 8, Annex II and Annex V);
b)   generate data sets appropriate for monitoring and reporting of Natura 2000 sites designated under the Habitats Directive and the Birds Directive;
c)   provide information supporting other environmental reporting requirements (e.g. in relation to biodiversity or environmental impact assessment);
d)   support lake management and restoration initiatives.
This European Standard:
a)   defines the key term of ‘hydromorphology’ and other terms relating to the physical characteristics of lakes and their hydrological regimes;
b)   details essential features and processes of lakes that should be characterised as part of a hydromorphological survey and for determining the hydromorphological condition of a lake;
c)   identifies and defines the key pressures affecting European lakes;
d)   provides guidance on strategies for collecting hydromorphological data depending on resources available and the anticipated use of the assessment; a hierarchy of approaches is recognised from the ‘overview method’ utilising existing databases, maps and remote sensing data through to recognised field-based survey techniques such as Lake Habitat Survey (LHS);
e)   offers guidance on data presentation;
f)   establishes guidance on data quality assurance issues.
This European Standard does not deal with biological assessments in lakes such as the presence or absence of individual species or community composition, nor does it attempt to link specific hydromorphological features with their associated biological communities or to create a classification based on such links. However, it is relevant where plants or other organisms form significant structural elements of the habitat (e.g. a gradation from riparian to littoral vegetation).
With respect to the WFD, the hydromorphological condition of a lake only contributes to its status classification at high ecological status (HES). Hydromorphological conditions are not defined for good and moderate status but shall be sufficient to support the biological elements.

This European Standard gives guidance for sampling, preservation, storage, quantification and qualitative analysis of phytoplankton from marine waters. Guidance for quantification is limited to the use of light microscopy with phase contrast and epifluorescence.
This European Standard specifies:
- the development of the sampling programme;
- requirements for sampling equipment;
- procedures for sampling and treatment of samples in the field;
- methods for quantification;
- qualitative analysis.
This European Standard describes minimum requirements for environmental monitoring.

This document describes methods for developing and applying Multimetric Indices used for assessing rivers, lakes, transitional waters or wetlands. It is suitable for use with data on fish, benthic invertebrates, macrophytes, phytoplankton, and phytobenthos.

This European Standard provides guidelines for the survey/sampling, identification and basic quantification of phytobenthos (other than macrophytes) in running waters. It is applicable to rivers where benthic algae and bryophytes are the main phototrophs. This method encompasses all phytobenthic growth forms and enables biological responses to environmental events over one or more years to be monitored. In this respect it provides an alternative to methods based on benthic diatoms (EN 13946; EN 14407) and macrophytes (EN 14184). Data obtained for the phytobenthos growth forms are suitable for pilot surveys, water quality assessment and trend monitoring. This European Standard encompasses all aspects from the design of survey and sampling programmes to the identification and basic quantification of the phytobenthos.

This guidance standard defines a method for surveying aquatic macrophytes in lakes - primarily for the purpose of assessing ecological status, using these organisms as an element of biological quality. The information provided by this method includes the composition and abundance of the aquatic macrophyte flora.
For a complete assessment of ecological status, other elements of biological quality should also be assessed.
The general principle of the approach described in this European Standard may also form the basis for the monitoring and assessment of macrophytes in lakes, for example, for conservation purposes.

The procedure described in this European Standard is based on the standard settling technique as defined by Utermöhl in 1958 [31]. It describes a general procedure for the estimation of abundance and taxonomic composition of marine and freshwater phytoplankton by using inverted light microscopy and sedimentation chambers, including the preceding steps of preservation and storage. Emphasis is placed on optimizing the procedure for the preparation of the microscopic sample. Many of the general principles of the approach described may also be applied to other techniques of enumerating algae (or other entities) using a (conventional) microscope, some of which are described in Annex E. This guidance standard does not cover field collection of samples or the analysis of picoplankton, quantitative analysis of free-floating mats of Cyanobacteria or specific preparation techniques for diatoms.

This guidance standard specifies equipment and procedures for collecting floating pupal exuviae of Chironomidae from aquatic habitats; rivers from source to estuary, canals, ponds, lakes and sea coasts. Guidance in preparing specimens for subsequent identification is provided. These samples provide representative data on relative species abundance, suitable for numerical analysis, classification and monitoring of environmental conditions.

This guidance standard defines activities appropriate for ensuring that the quality of ecological assessments in surface waters (including rivers, lakes, transitional and coastal waters and the open sea) and sediments meets specified requirements. This standard also covers hydromorphological aspects relevant to ecological assessment. While it has particular importance in relation to the assessment of ecological status in surface waters, it also applicable to other types of investigation and habitat.

This guidance standard describes general procedures for surveying zooplankton in standing waters for the purposes of water quality assessment and determination of ecological status.
Guidance on sampling procedures and the subsequent steps for preservation and storage are given. The sampling procedures provide estimate for species occurrence and their abundance (relative or absolute), including spatial distribution and temporal trends, for a given body of water. Calculation of biomass and production is made possible.
This method is restricted to the sampling of multicellular zooplankton that inhabit the pelagic and littoral regions of lakes, reservoirs and ponds. The sampling procedure may be also employed in slow running waters and canals.
NOTE   The field methods described are suitable for the collection of open-water plankton and littoral plankton species. They are inappropriate for the collection of littoral species that primarily live on or in the surface of sediments and on the surface of aquatic plants.

ISO 5667-19:2004 provides guidance for the sampling of sediments in marine areas for analyses of their physical and chemical properties for monitoring purposes and environmental assessments. It encompasses sampling strategy, requirements for sampling devices, observations made and information obtained during sampling, handling sediment samples, and packaging and storage of sediment samples.
ISO 5667-19:2004 does not provide guidelines for data treatment and analysis.
ISO 5667-19:2004 is not intended to give guidance for sampling of freshwater sediments.

This European Standard provides procedures to be used by trained persons in evaluating fish communities in streams, rivers and littoral areas for the purpose of classification of ecological status. These procedures allow standardisation of sampling methods for descriptions of fish communities. The use of standardised methods is a critical requirement for the comparability of results.
This European Standard describes an electric fishing method to be used when catching fish for the purpose of characterising species richness, composition, abundance and age structure of a given fish community. Sampling-related issues include obtaining permissions, concerns about endangered species, protective measures of importance for the user of the sampling apparatus and co-ordination of activities with other sampling programmes.
The processing of samples covers taxonomic identification, counting, measurement of biologic parameters (length, weight etc.), and examination of fish for external anomalies.

Migrated from Progress Sheet (TC Comment) (2000-07-10): TC N 315 (work programme 980901): UAP on ISO 9439 (to be published) TA/980916

Migrated from Progress Sheet (TC Comment) (2000-07-10): Splitting of WI 00230095 into WIs 095 and 108 (TC Res. 146/1996) ++ N315: New TD (TA/980915)

Migrated from Progress Sheet (TC Comment) (2000-07-10): Result of the splitting of WI 095 ++ N 315: New TD (980916)

Migrated from Progress Sheet (TC Comment) (2000-07-10): N 315: New TD (980916)

This document specifies a method for the sampling and laboratory preparation of benthic diatoms for ecological status and water quality assessments. The sampling and preparation procedures described can be used for later investigations using either light microscopy or molecular methods. Data produced by this method are suitable for production of indices based on the relative abundance of taxa.
Analysis using molecular methods is not within the scope of the document.

This document describes methods for data sampling and procedures for data evaluation, of fish populations in large rivers and lakes, using hydroacoustic equipment. This document provides information on routines for data sampling, data handling and reporting. A selected literature with references in support of this document is given in the bibliography.

This proposed guidance standard provides guidelines for the survey/sampling, identification and basic quantification of phytobenthos (other than macrophytes) in running waters. It is applicable to rivers where benthic algae and bryophytes are the main phototrophs.  This method encompasses all phytobenthic growth forms and enables biological responses to environmental events over one or more years to be monitored.  In this respect it provides an alternative to methods based on benthic diatoms (EN 13946; EN 14407) and macrophytes (EN 14184).  Data obtained for the phytobenthos growth forms are suitable for pilot surveys, water quality assessment and trend monitoring.  This proposed European Standard encompasses all aspects from the design of survey and sampling programmes to the identification and basic quantification of the phytobenthos.

This document provides guidance on the features to be recorded when characterising 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 survey methods, data processing, interpretation and presentation of results. Whilst it has particular importance in relation to the reporting requirements of the WFD, it also has considerably wider scope for other applications. Although hydromorphology is dependent on hydrology and underlying geology, this standard is focused on the structural features of rivers and on river continuity. In addition, whilst recognising the important influence of hydromorphology on plant and animal ecology and, conversely, the influence of plants and animals on hydromorphology, no attempt is made to provide guidance in this area.

This document establishes methods for the identification and enumeration of relative proportions of diatom taxa on prepared slides and of data interpretation relevant to assessments of water quality in rivers and streams. It is suitable for use with indices and assessment methods based on the relative abundance of taxa. The methods for identification and enumeration can also be applied to the study of benthic diatoms in other habitats provided that data interpretation methods appropriate to these habitats are used.

This European Standard specifies a method for surveying aquatic macrophytes in running waters for the purpose of assessing ecological status, using these organisms as elements of biological quality. The information provided by this method includes the composition and abundance of the aquatic macrophyte flora.
The general principles of the approach described in this European Standard may also be applied when monitoring water bodies in the fluvial corridor of a river, such as side channels and oxbows.
It is recognised that for a complete assessment of ecological status, other elements of biological quality should also be assessed.

This guidance standard establishes a method for sampling and laboratory preparation of benthic diatoms for water quality assessments and studies of temporal change in running waters. It is suitable for use with indices based on the relative abundance of taxa. With appropriate modifications the method can be applied to the study of benthic diatoms in lakes.

ISO 16665:2005 provides guidelines on the quantitative collection and processing of subtidal soft-bottom macrofaunal samples in marine waters.
ISO 16665:2005 encompasses: development of the sampling programme; requirements for sampling equipment; sampling and sample treatment in the field; sorting and species identification; storage of collected and processed material.
ISO 16665:2005 does not specifically address the following, although some elements may be applicable: bioassay sub-sampling; deep water (> 750 m) or offshore sampling; in situ faunal studies, e.g. recolonisation assays; nonbenthic organisms caught in the sampling device; estuarine sampling; intertidal sampling; meiofaunal sampling and analysis; sampling by dredge and sledge; Self-Contained Underwater Breathing Apparatus (SCUBA) sampling; statistical design.
Accuracy of position fixing is determined by the geographical area, equipment used and survey objective.