1、July 2014Translation by DIN-Sprachendienst.English price group 12No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 13
2、.060.70!%5xB“2188531www.din.deDDIN EN 13946Water quality Guidance for the routine sampling and preparation of benthic diatomsfrom rivers and lakes;English version EN 13946:2014,English translation of DIN EN 13946:2014-07Wasserbeschaffenheit Anleitung zur Probenahme und Probenaufbereitung von benthis
3、chen Kieselalgen ausFliegewssern und Seen;Englische Fassung EN 13946:2014,Englische bersetzung von DIN EN 13946:2014-07Qualit de leau Guide pour lchantillonnage en routine et le prtraitement des diatomes benthiques derivires et de plans deau;Version anglaise EN 13946:2014,Traduction anglaise de DIN
4、EN 13946:2014-07SupersedesDIN EN 13946:2003-09www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 21 pages06.14 DIN EN 13946:2014-07 2 A comma is used as the decimal marker. National forewordThis document (EN 13946:2014) has been prepared b
5、y Technical Committee CEN/TC 230 “Water analysis” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committee), Working Committee NA 119-01-03-05-06 AK Biologisch-kologische Gews-seruntersuchung of NA 11
6、9-01-03 AA Wasseruntersuchung. Designation of the method: Guidance for the routine sampling and preparation of benthic diatom samples from rivers and lakes (M 13): Method DIN EN 13946 M 13 This standard has been published to implement the Water Framework Directive (WFD), Directive 2000/60/EC of the
7、European Parliament and of the Council of 23 October 2000 establishing a framework for Community action in the field of water policy1). 1) Registered in the DITR database of DIN Software GmbH, obtainable from: Beuth Verlag GmbH, 10772 Berlin. DIN EN 13946:2014-07 3 Expert assistance and specialized
8、laboratories will be required to perform the analyses described in this standard. Existing safety requirements are to be observed. Depending on the objective of the analysis, a check shall be made on a case-by-case basis as to whether and to what extent additional conditions will have to be specifie
9、d. This standard has been prepared by the Normenausschuss Wasserwesen (Water Practice Standards Com-mittee) in collaboration with the Wasserchemische Gesellschaft Fachgruppe in der Gesellschaft Deutscher Chemiker (Water Chemistry Society Division of the German Chemical Society). It is part of the se
10、ries Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung (German standard methods for the examination of water, waste water and sludge): Guidance for the routine sampling and preparation of benthic diatom samples from rivers and lakes (M 13): Standard methods published as DIN S
11、tandards are obtainable from Beuth Verlag GmbH, either individually or grouped in volumes. The standard methods included in the loose-leaf publication entitled Deutsche Einheits-verfahren zur Wasser-, Abwasser- und Schlammuntersuchung will continue to be published by Beuth Verlag GmbH and Wiley-VCH
12、Verlag GmbH b) the scope of this European Standard has been expanded to include lakes; c) the standard has been editorially revised. Previous editions DIN EN 13946: 2003-09 EN 13946 March 2014 ICS 13.060.70 Supersedes EN 13946:2003English Version Water quality - Guidance for the routine sampling and
13、 preparation of benthic diatoms from rivers and lakes Qualit de leau - Guide pour lchantillonnage en routine et le prtraitement des diatomes benthiques de rivires et de plans deau Wasserbeschaffenheit - Anleitung zur Probenahme und Probenaufbereitung von benthischen Kieselalgen aus Fliegewssern und
14、Seen This European Standard was approved by CEN on 20 December 2013. CEN members are bound to comply with the CEN/CENELEC 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 bibliographic
15、al references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to 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
16、of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of M
17、acedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey andUnited Kingdom. CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 C
18、EN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 13946:2014 EEUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 13946:2014 (E) 2 Content
19、s Page Foreword 3 Introduction .4 1 Scope 5 2 Terms and definitions .5 3 Principle 6 4 Equipment 6 4.1 Equipment for field sampling .6 4.2 Laboratory equipment .6 5 Reagents .6 6 Procedure .8 6.1 Choice of substratum 8 6.2 Sample site selection 8 6.3 Sampling methods .8 6.3.1 Moveable natural hard s
20、urfaces .8 6.3.2 Method for sampling vertical man-made surfaces in situ .9 6.3.3 Use of artificial substrata . 10 6.3.4 Sample collection from submerged macrophytes and macroalgae 10 6.3.5 Sample collection from emergent macrophytes . 10 6.4 Preparation prior to microscopic examination 11 6.4.1 Pres
21、ervation and preliminary laboratory treatment 11 6.4.2 Methods for cleaning diatoms . 11 6.4.3 Preparation of permanent slides . 11 Annex A (informative) Methods for cleaning diatoms for microscopic examination 13 A.1 General . 13 A.2 Method 1: Hot hydrogen peroxide 13 A.2.1 Apparatus 13 A.2.2 Reage
22、nts 13 A.2.3 Procedure 13 A.3 Method 2: Cold hydrogen peroxide 14 A.3.1 Apparatus and reagents . 14 A.3.2 Procedure 14 A.4 Method 3: Hot hydrogen peroxide with potassium dichromate 14 A.4.1 Apparatus 14 A.4.2 Reagents 14 A.4.3 Procedure 14 A.5 Method 4: Cold acid (permanganate) method of cleaning .
23、15 A.5.1 Apparatus 15 A.5.2 Reagents 15 A.5.3 Procedure 15 Bibliography . 17 DIN EN 13946:2014-07EN 13946:2014 (E) 3 Foreword This document (EN 13946:2014) has been prepared by Technical Committee CEN/TC 230 “Water analysis”, the secretariat of which is held by DIN. This European Standard shall be g
24、iven the status of a national standard, either by publication of an identical text or by endorsement, at the latest by September 2014, and conflicting national standards shall be withdrawn at the latest by September 2014. Attention is drawn to the possibility that some of the elements of this docume
25、nt may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. This document supersedes EN 13946:2003. This document contains the following technical changes in comparison with the previous edition: this European Standard is no
26、w also applicable for the sampling of benthic diatoms in lakes. 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, Esto
27、nia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 13946:2014-07EN
28、13946:2014 (E) 4 Introduction Diatoms are an important component of aquatic ecosystems and constitute a water quality monitoring tool where the primary objective is either a measure of ecological status based on diatoms as one compartment of the ecosystem or the impact of specific components of wate
29、r quality (e.g. eutrophication, acidification). The requirement for the monitoring of such processes is inherent in the Water Framework Directive (2000/60/EC) 7 and Urban Waste Water Treatment Directive (91/271/EEC) 8 in addition to other EU Directives and international agreements. This European Sta
30、ndard covers aspects of sampling and preparation relevant to assessment of water quality and ecological status using benthic diatoms. These sampling instructions will result in samples suitable for quantifying relative numbers of benthic diatom taxa present. If it is necessary to quantify absolute n
31、umbers of taxa, or fresh weight per unit area, modifications to the method are required, which are not within the scope of this European Standard. The use of diatoms as indicators of river and lake quality is widely accepted both in Europe and the USA. The methodology is based on the fact that all d
32、iatom species have tolerance limits and optima with respect to their preference for environmental conditions such as nutrients, organic pollution and acidity. Polluted waters will tend to support an increased abundance of those species whose optima correspond with the levels of the pollutant in ques
33、tion. Conversely, certain species are intolerant of elevated levels of one or more pollutants, whilst others may occur in a wide range of water qualities. Methods using diatoms to assess water quality have been developed in several European countries (recent work is summarized in the proceedings of
34、three symposia 1 to 3. The methodologies for evaluating the diatom data vary but the sampling and preparation processes are similar 5, 6. According to the precise usage to which this European Standard is to be put it is essential for specifiers and users to mutually agree on any necessary variations
35、 or optional procedural details prior to use. All numerical values given in this standard are approximate. WARNING Persons using this European Standard should be familiar with usual laboratory practice. This European Standard does not purport to address all of the safety problems, if any, associated
36、 with its use. It is the responsibility of the user to establish appropriate health and safety practices and to ensure compliance with any national regulatory conditions. DIN EN 13946:2014-07EN 13946:2014 (E) 5 1 Scope This European Standard specifies a method for the sampling and laboratory prepara
37、tion 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. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply
38、. 2.1 artificial substratum introduced substratum substratum introduced into river or lake by operator specifically for colonization by diatoms 2.2 benthic diatoms diatoms living on natural or artificial substrata, rather than suspended in the water column 2.3 boulder mineral substratum with a diame
39、ter 256 mm 2.4 cobble mineral substratum with a diameter 64 mm and 256 mm 2.5 ecological status measure of the structure and functioning of aquatic communities 2.6 euphotic zone part of the water column in which there is sufficient light for photosynthesis 2.7 frustule cell wall of diatoms, composed
40、 of silica and consisting of two valves linked by two or more girdle bands 2.8 habitat specific environment in which an organism lives 2.9 pebble mineral substratum with a diameter 16 mm and 64 mm 2.10 riffle shallow part of a stream with swift flow, usually with a broken surface 2.11 substratum nat
41、ural or non-natural material from which benthic diatoms are sampled DIN EN 13946:2014-07EN 13946:2014 (E) 6 2.12 taxa taxonomic units, for example families, genera or species 2.13 valve structural component of the diatom frustule 3 Principle Benthic diatoms from submerged hard surfaces or submerged
42、macrophytes in rivers, streams or littoral zones of lakes are sampled in order to produce representative collections of the diatom assemblage indicative of water quality. Samples are cleaned using strong oxidizing agents in order to prepare diatoms for identification and enumeration. The data obtain
43、ed from the microscopic analysis of these samples are suitable for the production of diatom based water quality indices (see 1, 2, and 3). 4 Equipment 4.1 Equipment for field sampling 4.1.1 Appropriate water safety equipment 4.1.2 Waders 4.1.3 Stiff toothbrush (or other similar instrument) or knife
44、(or other suitable blade). 4.1.4 Plastic tray (approximately 30 cm 20 cm or larger). 4.1.5 Sample bottles with a tight fitting lid. 4.1.6 Indelible marker pen (or other means of labelling samples). If labels are used, these shall be capable of withstanding wet conditions. 4.1.7 Hoe, with a fine-mesh
45、ed net attached, attached to long handle (if vertical hard surfaces are to be sampled). 4.1.8 Glass-bottomed box or bucket (“Aquascope”) used for finding suitable substrata under some circumstances. 4.1.9 Plastic bags 4.2 Laboratory equipment See Annex A. 5 Reagents Reagents used in the preparation
46、of the diatom frustules need not be of analytical grade but should be of a quality appropriate for the digestion process. DIN EN 13946:2014-07EN 13946:2014 (E) 7 5.1 Preservatives Preservatives are required to stop cell division of diatoms and decomposition of organic matter. No preservative is nece
47、ssary if the sample is to be processed within a few hours of collection, as long as steps are taken to minimize cell division (i.e. by storage in cool, dark place). Lugols iodine can be used for short-term storage; however, it is not suitable for long-term storage, due to problems caused by sublimat
48、ion. Buffered formaldehyde or ethanol are recommended for long-term storage of samples. Samples may also be deep-frozen. 5.1.1 Buffered formaldehyde solution, HCHO, volume fraction at least 4 %. Dilute a stock solution of formaldehyde to a volume fraction of 4 % in a solution buffered to pH 7. Suita
49、ble buffers include HEPES (2-4-(2-hydroxyethyl)piperazin-1-ylethanesulfonic acid), borate and hexamethylene-tetramine. A final solution with a volume fraction of 1 % to 4 % in the sample is recommended. The quantity required will depend upon the amount of organic matter present in the sample. NOTE The buffer is necessary to prevent dissolution of the silica frustules. This is only necessary in alkaline waters, as diatoms dissolve in alkaline wa
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