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1、BRITISH STANDARD BS EN ISO 10705-2:2001 BS 6068-4.13:2000 Incorporating Corrigendum No. 1 Incorporating Amendment No. 1 to BS 6068-4.13:2000 (renumbers the BS as BS EN ISO 10705-2:2001) Water quality Detection and enumeration of bacteriophages Part 2: Enumeration of somatic coliphages The European S
2、tandard EN ISO 10705-2:2001 has the status of a British Standard ICS 07.100.20 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS EN ISO 10705-2:20001 This British Standa
3、rd, having been prepared under the direction of the Health and Environment Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 April 2000 BSI 19 November 2001 ISBN 0 580 34695 1 National foreword This British Standard is the official English lan
4、guage version of EN ISO 10705-2:2001 (as corrected 2001-11-07). It is identical with ISO 10705-2:2000. The UK participation in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee EH/3/4, Microbiological methods, which has the responsibility to: A list of organiz
5、ations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards
6、 Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard d
7、oes not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European dev
8、elopments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO foreword page, the ISO title page, pages ii to iv, pages 1 to 16, the annex ZA page and a back cover. The BSI copyright date displayed in this doc
9、ument indicates when the document was last issued. Amendments issued since publication Amd. No. DateComments 10977 Corrigendum June 2000Correct text for title added. 13447November 2001 Implementation of the European Standard Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2
10、006, Uncontrolled Copy, (c) BSI EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10705-2 August 2001 ICS 07.100.20 English version Water quality - Detection and enumeration of bacteriophages - Part 2: Enumeration of somatic coliphages (ISO 10705-2:2000) Qualit de leau - Dtection et dnombreme
11、nt des bactriophages - Partie 2: Dnombrement des coliphages somatiques (ISO 10705-2:2000) Wasserbeschaffenheit - Nachweis und Zhlung von Bakteriophagen - Teil 2: Zhlung von somatischen Coliphagen (ISO 10705-2:2000) This European Standard was approved by CEN on 22 June 2001. CEN members are bound to
12、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 application to the Managem
13、ent 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 Management Centre has the same status as the off
14、icial versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMI
15、T EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: rue de Stassart, 36 B-1050 Brussels 2001 CENAll rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 10705-2:2001 E Licensed Copy: sheffieldun sheffieldun, na, Fri
16、 Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI EN ISO 10705-2:2001 (E) CORRECTED 2001-11-07 Foreword The text of the International Standard from Technical Committee ISO/TC 147 “Water quality“ of the International Organization for Standardization (ISO) has been taken over as an European
17、Standard 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 by endorsement, at the latest by February 2002, and conflicting national standa
18、rds shall be withdrawn at the latest by February 2002. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Icelan
19、d, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. NOTE: Normative references to International Standards are listed in annex ZA (normative). Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy
20、, (c) BSI Reference number ISO 10705-2:2000(E) INTERNATIONAL STANDARD ISO 10705-2 First edition 2000-03-01 Water quality Detection and enumeration of bacteriophages Part 2: Enumeration of somatic coliphages Qualit de leau Dtection et dnombrement des bactriophages Partie 2: Dnombrement des coliphages
21、 somatiques Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ii? EN ISO 107052:2001 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI iii ContentsPage 1Scope 1 2Normative references1 3T
22、erms and definitions .1 4Safety precautions.2 5Principle2 6Diluent, culture media and reagents2 6.1Basic materials.2 6.2Diluent.2 7Apparatus and glassware.2 8Microbiological reference cultures4 9Sampling.4 10Preparation of test material5 10.1Culturing and maintenance of host strains 5 10.2Calibratio
23、n of absorbance measurements for counts of viable microorganisms6 11Procedure.6 11.1Preparation of inoculum cultures 6 11.2Standard procedure.7 11.3Method for samples with high bacterial background flora .7 11.4Samples with low phage counts 7 11.5Presence/absence test8 11.6Quality assurance8 12Expre
24、ssion of results 9 12.1Plaque count procedures (11.2 to 11.4).9 12.2Presence/absence test (11.5)9 13Test report10 Annex A (normative) Culture media, reagents and diluent.11 Annex B (informative) General description of somatic bacteriophages .14 Annex C (informative) Culturing of bacteriophage ? ? ?
25、?X174.15 Bibliography16 EN ISO 107052:2001 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI iv Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The w
26、ork of preparing International Standards is normally carried out through ISO technical committees. Each member body interested 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-govern
27、mental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 3. Dra
28、ft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this part
29、 of ISO 10705 may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. InternationalStandardISO 10705-2waspreparedbyTechnicalCommitteeISO/TC 147,Waterquality, Subcommittee SC 4,Microbiological methods. ISO 10705 consists of the following p
30、arts, under the general titleWater quality Detection and enumeration of bacteriophages: ?Part 1: Enumeration of F-specific RNA bacteriophages ?Part 2: Enumeration of somatic coliphages ?Part 3: Concentration methods ?Part 4: Enumeration of bacteriophages infectingBacteroides fragilis Annex A forms a
31、 normative part of this part of ISO 10705. Annexes B and C are for information only. EN ISO 107052:2001 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 1 Water quality Detection and enumeration of bacteriophages Part 2: Enumeration of somati
32、c coliphages 1Scope This part of ISO 10705 specifies a method for the detection and enumeration of somatic coliphages by incubating the sample with an appropriate host strain. The method is applicable to all kinds of water, sediments and sludge extracts, where necessary after dilution. The method is
33、 also applicable to shellfish extracts. In the case of low phage numbers, a preconcentration step may be necessary for which a separate International Standard will be developed. NOTEIt is desirable for International Standards to be adopted as widely as possible. This part of ISO 10705 includes refer
34、ence to alternative procedures which obviate the need for expensive materials or equipment which may not be readily available in developing countries. Use of these alternatives will not affect the performance of this method. 2Normative references The following normative documents contain provisions
35、which, through reference in this text, constitute provisions of this part of ISO 10705. For dated references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this part of ISO 10705 are encouraged to investigate the possibilit
36、y of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain registers of currently valid International Standards. ISO 31-0:1992,Quantities and units Part 0
37、: General principles. ISO 3696:1987,Water for analytical laboratory use Specification and test methods. ISO 5667-1:1980,Water quality Sampling Part 1: Guidance on the design of sampling programmes. ISO 5667-2:1991,Water quality Sampling Part 2: Guidance on sampling techniques. ISO 5667-3:1994,Water
38、quality Sampling Part 3: Guidance on the preservation and handling of samples. ISO 6887:1983,Microbiology General guidance for the preparation of dilutions for microbiological examination. ISO 8199:1988,Water quality General guide to the enumeration of micro-organisms by culture. ISO/IEC Guide 2,Sta
39、ndardization and related activities General vocabulary. 3Terms and definitions For the purposes of this part of ISO 10705, the terms and definitions given in ISO/IEC Guide 2 and the following apply. EN ISO 107052:2001 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Un
40、controlled Copy, (c) BSI 2 3.1 somatic coliphage bacterial virus which is capable of infecting selectedEscherichia colihost strains (and related strains) by attachment to the bacterial cell wall as the first step of the infection process NOTESomatic coliphages produce visible plaques (clearance zone
41、s) in a confluent lawn of host bacteria grown under appropriate culture conditions. 4Safety precautions The host strain used in this standard is non-pathogenic to man and animals, and should be handled in accordance with the normal (national or international) safety procedures for bacteriological la
42、boratories. Somatic coliphages are also non-pathogenic to man and animals, but some types are very resistant to drying. Appropriate precautions should therefore be taken to prevent cross-contamination of test materials, particularly when examining or handling cultures of high titre or when inoculati
43、ng cultures of the host strain. Such procedures shall be carried out in a biohazard cabinet or a separate area of the laboratory. Chloroform is a carcinogenic substance. Observe relevant safety precautions or use an alternative method of equal efficacy. 5Principle The sample is mixed with a small vo
44、lume of semi-solid nutrient medium. A culture of host strain is added and plated on a solid nutrient medium. After this, incubation and reading of plates for visible plaques takes place. The results are expressed as the number of plaque-forming particles, pfp (also termed plaque-forming units, pfu),
45、 per unit of medium volume. 6Diluent, culture media and reagents 6.1Basic materials Use ingredients of uniform quality and chemicals of analytical grade for the preparation of culture media and reagents and follow the instructions given in annex A. For information on storage see ISO 8199, except whe
46、re indicated in this part of ISO 10705. Alternatively, use dehydrated complete media and follow strictly the manufacturers instructions. For the preparation of media, use glass-distilled water or deionized water free from substances which might inhibit bacterial growth under the conditions of the te
47、st, and complying with ISO 3696. NOTEUse of other grades of chemicals is permissible providing they are shown to be of equal performance in the test. 6.2Diluent For making sample dilutions, use peptone-saline solution (A.7) or another diluent complying with ISO 6887. 7Apparatus and glassware Usual m
48、icrobiological laboratory equipment, including 7.1Hot-air oven for dry-heat sterilization and an autoclave. Apart from apparatus supplied sterile, glassware and other equipment shall be sterilized according to the instructions given in ISO 8199. 7.2Incubator or water bath, thermostatically controlle
49、d at (36 ? 2) C. EN ISO 107052:2001 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 09:53:09 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 3 7.3Incubator or water bath, thermostatically controlled at (36 ? 2) C and equipped with a shaking device, for example a rotating platform at (100 ? 10) r/min. 7.4Water bath or heating block, thermostatically controlled at (45 ? 1) C. 7.5Water bath or equivalent device for melting of agar media. 7.6pH meter. 7.7Counting apparatus with indirect, oblique light. 7.8Deep freezer,