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 LAWBS EN ISO 10705-2:20001 This British Standard, having been prepared under the direction of the Health and Environment Sector Committee, was published u
3、nder 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 language version of EN ISO 10705-2:2001 (as corrected 2001-11-07). It is identical with ISO 10705-2:2000. The U
4、K 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 organizations represented on this committee can be obtained on request to its secretary. Cross-references The Briti
5、sh 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 Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British
6、 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 does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to t
7、he responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside f
8、ront 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 document indicates when the document was last issued. Amendments issued since publication Amd. No. Date Comment
9、s 10977 Corrigendum June 2000 Correct text for title added. 13447 November 2001 Implementation of the European StandardEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM ENISO107052 August2001 ICS07.100.20 Englishversion WaterqualityDetectionandenumerationofbacteriophages Part2:Enumerationofsomaticcolip
10、hages(ISO107052:2000) QualitdeleauDtectionetdnombrementdes bactriophagesPartie2:Dnombrementdescoliphages somatiques(ISO107052:2000) WasserbeschaffenheitNachweisundZhlungvon BakteriophagenTeil2:Zhlungvonsomatischen Coliphagen(ISO107052:2000) ThisEuropeanStandardwasapprovedbyCENon22June2001. CENmember
11、sareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcernings uchnational standardsmaybeobtainedonapplicationtotheManagementCentreortoanyCENmember.
12、ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheManagementCentrehasthesamestatusasthe official versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,
13、CzechRepublic,Denmark,Finland,France,Germany,Greece, Iceland,Ireland,Italy,Luxembourg,Netherlands,Norway,Portugal,Spain,Sweden,SwitzerlandandUnitedKingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITEEFRNORMUNG ManagementCentre:ruedeStassart,36B1050Brussels 2001C
14、EN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.ENISO107052:2001EENISO107052:2001(E) CORRECTED20011107 Foreword Thete xto ftheI nternational Standard from TechnicalCo mmittee ISO/TC147 “Waterquali ty“ oftheInternationalOrganizationforStandardization(IS
15、O)hasbeentakenoverasan EuropeanStandardbyTechnicalCommitteeCEN/TC230“Wateranalysis“,thesecretariatof whichisheldbyDIN. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublication ofanidenticaltextorbyendorsement,atthelatestbyFebruary2002,andconflictingnational standardsshallbewi
16、thdrawnatthelatestbyFebruary2002. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsof thefollowingcountriesareboundtoimplementthisEuropeanStandard:Austria,Belgium, CzechRepublic,Denmark,Finland,France,Germany,Greece,Iceland,Ireland,Italy, Luxembourg,Netherlands,Norway,Po
17、rtugal,Spain,Sweden,SwitzerlandandtheUnited Kingdom. NOTE:NormativereferencestoInternationalStandardsarelistedinannexZA(normative).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: Enumer
18、ation of somatic coliphages Qualit de leau Dtection et dnombrement des bactriophages Partie 2: Dnombrement des coliphages somatiquesii ENISO107052:2001iii Contents Page 1 Scope 1 2 Normative references 1 3 Terms and definitions .1 4 Safety precautions.2 5 Principle2 6 Diluent, culture media and reag
19、ents2 6.1 Basic materials.2 6.2 Diluent.2 7 Apparatus and glassware .2 8 Microbiological reference cultures 4 9 Sampling.4 10 Preparation of test material 5 10.1 Culturing and maintenance of host strains 5 10.2 Calibration of absorbance measurements for counts of viable microorganisms 6 11 Procedure
20、 .6 11.1 Preparation of inoculum cultures 6 11.2 Standard procedure.7 11.3 Method for samples with high bacterial background flora .7 11.4 Samples with low phage counts 7 11.5 Presence/absence test 8 11.6 Quality assurance8 12 Expression of results 9 12.1 Plaque count procedures (11.2 to 11.4).9 12.
21、2 Presence/absence test (11.5)9 13 Test report 10 Annex A (normative) Culture media, reagents and diluent.11 Annex B (informative) General description of somatic bacteriophages .14 Annex C (informative) Culturing of bacteriophage X174.15 Bibliography16 ENISO107052:2001iv Foreword ISO (the Internatio
22、nal 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 committee has been
23、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 electrotechnical standar
24、dization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 3. Draft 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
25、 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this part of ISO 10705 may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. International Standard ISO 10705-2 was prepared b
26、y Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4, Microbiological methods. ISO 10705 consists of the following parts, under the general title Water quality Detection and enumeration of bacteriophages: Part 1: Enumeration of F-specific RNA bacteriophages Part 2: Enumeration of somat
27、ic coliphages Part 3: Concentration methods Part 4: Enumeration of bacteriophages infecting Bacteroides fragilis Annex A forms a normative part of this part of ISO 10705. Annexes B and C are for information only. ENISO107052:20011 Water quality Detection and enumeration of bacteriophages Part 2: Enu
28、meration of somatic coliphages 1 Scope 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 dilu
29、tion. The method is 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. NOTE It is desirable for International Standards to be adopted as widely as possible. This part of ISO
30、10705 includes reference 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. 2 Normative references The following normative document
31、s contain provisions 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 inves
32、tigate the possibility 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, Quant
33、ities and units Part 0: 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 technique
34、s. ISO 5667-3:1994, Water 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
35、culture. ISO/IEC Guide 2, Standardization and related activities General vocabulary. 3 Terms 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. ENISO107052:20012 3.1 somatic coliphage bacterial virus which is capabl
36、e of infecting selected Escherichia coli host strains (and related strains) by attachment to the bacterial cell wall as the first step of the infection process NOTE Somatic coliphages produce visible plaques (clearance zones) in a confluent lawn of host bacteria grown under appropriate culture condi
37、tions. 4 Safety 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 laboratories. Somatic coliphages are also non-pathogenic to man and animals, b
38、ut 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 inoculating cultures of the host strain. Such procedures shall be carried out in a bi
39、ohazard 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. 5P r i n c i p l e The sample is mixed with a small volume of semi-solid nutrient medium. A culture of host strain is adde
40、d 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), per unit of medium volume. 6 Diluent, culture media and reagents 6.
41、1 Basic 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 where indicated in this part of ISO 10705. Alternatively, use dehydra
42、ted 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 test, and complying with ISO 3696. NOTE Use of other grades of chemi
43、cals is permissible providing they are shown to be of equal performance in the test. 6.2 Diluent For making sample dilutions, use peptone-saline solution (A.7) or another diluent complying with ISO 6887. 7 Apparatus and glassware Usual microbiological laboratory equipment, including 7.1 Hot-air oven
44、 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. 7.2 Incubator or water bath, thermostatically controlled at (36 2) C. ENISO107052:20013 7.3 Incubator or water bath,
45、 thermostatically controlled at (36 2) C and equipped with a shaking device, for example a rotating platform at (100 10) r/min. 7.4 Water bath or heating block, thermostatically controlled at (45 1) C. 7.5 Water bath or equivalent device for melting of agar media. 7.6 pH meter. 7.7 Counting apparatu
46、s with indirect, oblique light. 7.8 Deep freezer, thermostatically controlled at ( 20 5) C. 7.9 Deep freezer, thermostatically controlled at ( 70 10) C or liquid nitrogen storage vessel. 7.10 Spectrophotometer, capable of holding cuvettes of 1 cm optical path length or side-arm of nephelometric flas
47、ks (7.17) and equipped with a filter for the range 500 nm to 650 nm with a maximum bandwidth of 10 nm. Usual sterile, microbiological laboratory glassware or disposable plasticsware according to ISO 8199 and including 7.11 Petri dishes of 9 cm or 14 cm to 15 cm diameter, vented. 7.12 Graduated pipet
48、tes of 0,1 ml, 1 ml, 5 ml and 10 ml capacity and Pasteur pipettes. 7.13 Glass bottles of suitable volume. 7.14 Culture tubes with caps or suitable alternative. 7.15 Measuring cylinders of suitable capacity. 7.16 Conical flasks of 250 ml to 300 ml capacity, with cotton wool plugs or suitable alternative. 7.17 Cuvettes of optical path length 10 mm or nephelometric conical flasks with cylindrical side-arms which f
