1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 15972:2011Water quality Guidance onquantitative and qualitativeinvestigations of marinephytoplanktonBS EN 15972:2011 BRITISH STANDARDNational forewordThis British Standard
2、is the UK implementation of EN 15972:2011.The UK participation in its preparation was entrusted to TechnicalCommittee EH/3/5, Biological Methods.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the neces
3、saryprovisions of a contract. Users are responsible for its correctapplication. BSI 2011ISBN 978 0 580 67914 8ICS 13.060.70Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Commi
4、ttee on 30 September 2011.Amendments issued since publicationDate Text affectedBS EN 15972:2011EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15972 September 2011 ICS 13.060.70 English Version Water quality - Guidance on quantitative and qualitative investigations of marine phytoplankton Quali
5、t de leau - Guide pour ltude quantitative et qualitative du phytoplancton marin Wasserbeschaffenheit - Anleitung fr die quantitative und qualitative Untersuchung von marinem Phytoplankton This European Standard was approved by CEN on 29 July 2011. CEN members are bound to comply with the CEN/CENELEC
6、 Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or t
7、o any CEN member. This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the offici
8、al versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slova
9、kia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of exploitation in any form and by any means reserved worldwide
10、for CEN national Members. Ref. No. EN 15972:2011: EBS EN 15972:2011EN 15972:2011 (E) 2 Contents Page Foreword 4Introduction .51 Scope 52 Normative references 63 Terms and definitions .64 Aim and strategy for sampling phytoplankton .74.1 Sampling programme 74.2 Types of surveys74.3 Sampling stations
11、84.4 Sampling depth 94.5 Sampling frequency and duration . 104.6 Parallel water samples . 105 Equipment . 105.1 Equipment for quantitative sampling . 105.2 Equipment for qualitative sampling 105.3 Sample bottles 115.4 Fixatives and preservatives . 116 Survey procedures . 126.1 Description of procedu
12、res . 126.2 Preparation 126.3 Defining the position of sampling stations 126.4 Operating the sampling device . 126.5 Sample fixation . 136.6 Samples for analysis of living material 136.7 Sample labelling and recording of additional information . 136.8 Sample storage . 146.9 Equipment maintenance 146
13、.10 Requirements for personnel 147 Species identification and sample processing 147.1 Species identification . 147.2 Methods for quantification . 157.3 Methods for qualitative analysis . 167.4 Determination of biomass 177.5 Qualification requirements for personnel responsible for identification . 17
14、7.6 Literature . 177.7 Reporting of result 177.8 Data storage 18Annex A (informative) Sampling and sample depth . 19A.1 General . 19A.2 Sampling depth . 19A.3 Integrated samples . 19A.4 Combined samples . 20Annex B (informative) Water samplers 21Annex C (informative) Plankton net samples 22Annex D (
15、informative) Preservatives 23BS EN 15972:2011EN 15972:2011 (E) 3 Annex E (informative) Counting chambers for quantification without concentration of the phytoplankton cells . 25E.1 Sedgwick Rafter 25E.2 Palmer Maloney 25E.3 Haemacytometre 25Annex F (informative) Utermhl method 26Bibliography 27BS EN
16、 15972:2011EN 15972:2011 (E) 4 Foreword This document (EN 15972:2011) has been prepared by Technical Committee CEN/TC 230 “Water analysis”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of an identical text or
17、by endorsement, at the latest by March 2012, and conflicting national standards shall be withdrawn at the latest by March 2012. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for i
18、dentifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
19、Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. BS EN 15972:2011EN 15972:2011 (E) 5 Introduction Investigations of phytoplankton are an impo
20、rtant part of marine environment monitoring. Phytoplankton responds rapidly to environmental changes (supply of nutrient salts, climate, light access etc.), and monitoring of occurrence, species composition and biomass may therefore in many cases be used to characterise the environment and the degre
21、e of impacts. The European Union Water Framework Directive therefore requires monitoring of marine phytoplankton as a part of assessments of ecological conditions. Investigations of phytoplankton are also included in monitoring programmes connected with other European directives (Urban Waste Water T
22、reatment Directive, Habitats Directive), international conventions and national regulations. Monitoring of harmful/toxic phytoplankton is carried out both for the aquaculture industry and in connection with authorities control of organisms for human consumption (shellfish). This requires uniform pro
23、cedures for collection and quantification of phytoplankton. Most principles for characterisation of environmental conditions based on phytoplankton require the use of quantitative methods, i.e. that the occurrence of species and quantities can be related to a known water mass. Data interpretation fu
24、rther requires information on the physical and chemical properties of the water body (supporting parameters). In order for environmental authorities to utilise the information, it is important that the investigations are comparable in space and time and that the data is of high scientific quality. T
25、his European Standard focuses on a limited selection of methods that can be documented precisely, reproduced and which have been in use for some time. This European Standard does not comprise chlorophyll determination. For chlorophyll determination see ISO 10260 1. WARNING Persons using this Europea
26、n Standard should be familiar with normal laboratory and fieldwork practice. This European Standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the users to establish appropriate safety and health practices and to ensure compli
27、ance with any national regulatory conditions. 1 Scope 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
28、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 fo
29、r environmental monitoring. BS EN 15972:2011EN 15972:2011 (E) 6 2 Normative references The following referenced documents are necessary for the application of this document. For dated references, only the cited edition applies. For undated references, the latest edition of the referenced document (i
30、ncluding any amendments) applies. EN 15204, Water quality Guidance standard on the enumeration of phytoplankton using inverted microscopy (Utermhl technique) 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 euphotic zone upper zone of a water
31、body which is penetrated by light, and where the primary production over one or more days is equal to or greater than the respiration, i.e. a positive net production NOTE Empirically, the euphotic zone is considered to extend down to the depth where 1 % of the surface light is present. 3.2 phytoplan
32、kton community of free-living, suspended, mainly photosynthetic organisms in aquatic systems comprising Cyanobacteria and microscopic algae EN 15204:2006, 3.10 3.3 phytoplankton biomass mass of living matter comprising phytoplankton NOTE The phytoplankton biomass can be expressed in terms of chlorop
33、hyll content, the carbon content of the algae (algal carbon) or the volume of the phytoplankton (biovolume). 3.4 qualitative sample phytoplankton sample collected by plankton net, i.e. a concentrated sample where the size of the phytoplankton is mainly larger than the mesh diameter NOTE Allows for r
34、ecordings of taxon/taxa occurring in low concentrations. 3.5 quantitative sample phytoplankton sample where a part of a body of water is enclosed so that the density of each taxon and their relative abundance per unit volume may be recorded 3.6 receiving water body water body that receives an input
35、of material, of either natural or anthropogenic origin NOTE The term often appears in the context of anthropogenic input, for example, effluent from municipal waste water outlets or industrial processed water. EN ISO 16665:2005, 2.4 BS EN 15972:2011EN 15972:2011 (E) 7 3.7 sampling station precise lo
36、cation where samples are collected NOTE A sampling station is defined by its geographical position (OS National Grid Reference, latitude, longitude), its depth (relative to chart data and normalised to mean low water as given in tide tables) and any other invariant of physical conditions. The statio
37、n is delineated using the given level of precision. In cases of doubt when revisiting sampling stations, emphasis should be placed on landmarks and water depth. EN ISO 16665:2005, 2.7 3.8 stratification two or more water bodies with different physical properties (temperature, salinity, density) lyin
38、g upon each other NOTE A thermocline separates the water bodies from each other. 3.9 thermocline boundary layer between two horizontally arranged water bodies with different physical properties which change dramatically in the vertical 3.10 water body body of water, standing or flowing; a lake, rive
39、r, or other collection of water 3.11 water transparency estimate of turbidity determined by recording the depth at which a white disc with black and white quarters lowered into the water is no longer visible from the surface 4 Aim and strategy for sampling phytoplankton 4.1 Sampling programme Before
40、 a survey is carried out, the sampling programme shall be defined according to the individual aims of the survey and the required precision of results and their intended use. Sampling methodology, number of sampling stations, sampling frequency and methodology for sample processing will vary between
41、 different types of investigations. During development of the programme, consideration shall be given to hydrographic conditions in the survey area, information about local pollution sources, knowledge from previous surveys and any other conditions that may be of importance for the phytoplankton com
42、position in the survey area. For long-term baseline studies, it should be ensured that methodological changes do not have any influence on the results of time series. 4.2 Types of surveys 4.2.1 Pilot survey Pilot surveys shall provide a general overview of the occurrence of phytoplankton and physica
43、l and chemical conditions in the water body of the survey area. In areas where larger scale environmental surveys or monitoring is required, there can be a need to carry out a pilot survey in advance. This pilot survey shall provide the necessary information for developing more specific sampling pro
44、grammes. The requirements for equipment, methodology and repeatability are usually relatively simple. The phytoplankton samples shall be collected by plankton net and/or water samplers. Hydrographic data shall be collected with a continuous BS EN 15972:2011EN 15972:2011 (E) 8 recording probe or by u
45、se of a water collector and subsequent analysis of sampled water. The phytoplankton samples are processed in such a way that the most important taxon/taxa can be identified and quantified. NOTE In this context, the most important taxon/taxa are those that are dominant in terms of numbers and/or biom
46、ass and taxon/taxa that have characteristic properties (indicators of eutrophication, producers of toxins etc.). 4.2.2 Baseline survey and monitoring of biological diversity Baseline surveys should be carried out for environmental monitoring and characterisation of biodiversity. Detailed analyses sh
47、ould be carried out on the relative species composition and abundance within the phytoplankton communities. The investigation shall provide the basis for characterising environmental conditions in the relevant areas in accordance with set criteria or by comparison with phytoplankton communities in o
48、ther representative areas. The requirements for scientific documentation and ability to be replicated are usually relatively high. If the baseline survey is carried out over time in one area, the investigation will provide a temporal description of natural variations of the phytoplankton communities
49、 and document any gradual changes (trend monitoring). The investigation shall be carried out at fixed stations and follow a carefully defined programme. At the start of investigations, in areas where the hydrographic conditions are insufficiently known, a preliminary investigation of the hydrographic conditions shall be carried out to identify the different water
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