1、Water quality - Determination of the growth inhibition effects of waste waters, natural waters and chemicals on the duckweed Spirodela polyrhiza - Method using a stock culture independent microbiotest (ISO 20227:2017)BS EN ISO 20227:2017BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd
2、1 15/05/2013 15:06EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 20227 July 2017 ICS 13.060.70 English Version Water quality - Determination of the growth inhibition effects of waste waters, natural waters and chemicals on the duckweed Spirodela polyrhiza - Method using a stock culture ind
3、ependent microbiotest (ISO 20227:2017) Qualit de leau - Dtermination des effets dinhibition sur la croissance de la lentille deau Spirodela polyrhizapar les eaux uses, les eaux naturelles et les produits chimiques - Mthode utilisant un bioessai miniaturis indpendant dune culture mre (ISO 20227:2017)
4、 Wasserbeschaffenheit - Bestimmung der wachstumshemmenden Wirkung von Abwssern, natrlichen Wssern und Chemikalien auf die Wasserlinsenart Spirodela polyrhiza - Verfahren mittels Stammkultur unabhngigem mikrobiologischem Test (ISO 20227:2017) This European Standard was approved by CEN on 9 May 2017.
5、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 bibliographical references concerning such national standards may be obtained on a
6、pplication 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 of a CEN member into its own language and notified to the CEN-CENELEC
7、 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 Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy,
8、Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre:
9、 Avenue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 20227:2017 ENational forewordThis British Standard is the UK implementation of EN ISO 20227:2017.The UK participation in its preparation wa
10、s entrusted to Technical Committee EH/3/5, Biological Methods.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application.
11、The British Standards Institution 2017Published by BSI Standards Limited 2017ISBN 978 0 580 92023 3ICS 13.060.70Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 3
12、1 August 2017.Amendments/corrigenda issued since publicationDate Text affectedBRITISH STANDARDBS EN ISO 20227:2017EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 20227 July 2017 ICS 13.060.70 English Version Water quality - Determination of the growth inhibition effects of waste waters, nat
13、ural waters and chemicals on the duckweed Spirodela polyrhiza - Method using a stock culture independent microbiotest (ISO 20227:2017) Qualit de leau - Dtermination des effets dinhibition sur la croissance de la lentille deau Spirodela polyrhizapar les eaux uses, les eaux naturelles et les produits
14、chimiques - Mthode utilisant un bioessai miniaturis indpendant dune culture mre (ISO 20227:2017) Wasserbeschaffenheit - Bestimmung der wachstumshemmenden Wirkung von Abwssern, natrlichen Wssern und Chemikalien auf die Wasserlinsenart Spirodela polyrhiza - Verfahren mittels Stammkultur unabhngigem mi
15、krobiologischem Test (ISO 20227:2017) This European Standard was approved by CEN on 9 May 2017. 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-d
16、ate lists and bibliographical 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 translatio
17、n 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 official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Fo
18、rmer Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZAT
19、ION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 20227:2017 EBS EN ISO 20227:2017EN ISO 2022
20、7:2017 (E) 3 European foreword This document (EN ISO 20227:2017) has been prepared by Technical Committee ISO/TC 147 “Water quality“ in collaboration with 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 na
21、tional standard, either by publication of an identical text or by endorsement, at the latest by January 2018 and conflicting national standards shall be withdrawn at the latest by January 2018. Attention is drawn to the possibility that some of the elements of this document may be the subject of pat
22、ent rights. CEN shall not be held responsible for identifying 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,
23、Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the Un
24、ited Kingdom. Endorsement notice The text of ISO 20227:2017 has been approved by CEN as EN ISO 20227:2017 without any modification. ISO 20227:2017(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Test organisms 36 Growth medium . 36.1 Preparatio
25、n of stock solutions . 46.2 Preparation of the final concentration of modified Steinberg medium . 47 Apparatus . 48 Reference chemicals . 59 Procedure. 59.1 Germination of the Spirodela polyrhiza turions 59.2 Tests on effluents (and waste waters) 59.2.1 Addition of concentrated growth medium to the
26、effluent sample . 59.2.2 Preparation of the effluent dilutions 69.2.3 Procedure . 69.3 Tests on chemical compounds . 79.3.1 Range finding test . 79.3.2 Definitive test 89.4 Filling of the test plate with the toxicant dilutions 99.4.1 General 99.4.2 Procedure . 99.5 Transfer of the germinated turions
27、 in the test cups 109.6 Photo of the multiwell at the start of the toxicity test 109.7 Incubation of the multiwell . 109.8 Photo of the multiwell at the end of the toxicity test 119.9 Measurement of the area of the first fronds . 1110 Data treatment Calculation of the growth inhibition .1111 Validit
28、y criterion .1212 Test sensitivity .1213 Test with reference chemicals .1214 Test report 15Annex A (informative) Spirodela polyrhiza stock culturing for turion production .16Annex B (informative) Sensitivity of the Spirodela polyrhiza microbiotest 17Annex C (informative) Performance data .19Bibliogr
29、aphy .20 ISO 2017 All rights reserved iiiContents PageBS EN ISO 20227:2017ISO 20227:2017(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Test organisms 36 Growth medium . 36.1 Preparation of stock solutions . 46.2 Preparation of the final conce
30、ntration of modified Steinberg medium . 47 Apparatus . 48 Reference chemicals . 59 Procedure. 59.1 Germination of the Spirodela polyrhiza turions 59.2 Tests on effluents (and waste waters) 59.2.1 Addition of concentrated growth medium to the effluent sample . 59.2.2 Preparation of the effluent dilut
31、ions 69.2.3 Procedure . 69.3 Tests on chemical compounds . 79.3.1 Range finding test . 79.3.2 Definitive test 89.4 Filling of the test plate with the toxicant dilutions 99.4.1 General 99.4.2 Procedure . 99.5 Transfer of the germinated turions in the test cups 109.6 Photo of the multiwell at the star
32、t of the toxicity test 109.7 Incubation of the multiwell . 109.8 Photo of the multiwell at the end of the toxicity test 119.9 Measurement of the area of the first fronds . 1110 Data treatment Calculation of the growth inhibition .1111 Validity criterion .1212 Test sensitivity .1213 Test with referen
33、ce chemicals .1214 Test report 15Annex A (informative) Spirodela polyrhiza stock culturing for turion production .16Annex B (informative) Sensitivity of the Spirodela polyrhiza microbiotest 17Annex C (informative) Performance data .19Bibliography .20 ISO 2017 All rights reserved iiiContents PageBS E
34、N ISO 20227:2017ISO 20227:2017(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body inter
35、ested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical
36、 Commission (IEC) on all matters of electrotechnical standardization. The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO docum
37、ents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held respons
38、ible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/ patents).Any trade name used in this document is information given
39、 for the convenience of users and does not constitute an endorsement. For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) princip
40、les in the Technical Barriers to Trade (TBT) see the following URL: www .iso .org/ iso/ foreword .html.This document was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods.iv ISO 2017 All rights reservedBS EN ISO 20227:2017ISO 20227:2017(E)ForewordISO (t
41、he International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical commit
42、tee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotech
43、nical standardization. The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted
44、 in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rig
45、hts. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/ patents).Any trade name used in this document is information given for the convenience of users and does not cons
46、titute an endorsement. For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) se
47、e the following URL: www .iso .org/ iso/ foreword .html.This document was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods.iv ISO 2017 All rights reserved ISO 20227:2017(E)IntroductionDuckweeds are free-floating higher water plants commonly used in eco
48、toxicological research for the assessment of the toxicity of waste waters, natural waters and chemicals (see ISO 20079 and References 6 to 11 and in particular plant protection products, see Reference 12).Duckweeds are fast growing plants, many of which have a cosmopolitan distribution, and they are
49、, hence, well suited as primary producers for hazard assessment of pollutants in freshwater environments.Contrary to terrestrial plants, for which bioassays can be started from the “dormant” life stages (seeds), toxicity tests with duckweeds require continuous culturing and maintenance of live stocks, with the inherent biological, technical and financial costs.A few duckweed species, however, produce dormant vegetative buds (turions) which can be stored for long periods of time, and which can be germinated on demand at
