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1、BRITISH STANDARD BS EN ISO 10272-1:2006 Microbiology of food and animal feeding stuffs Horizontal method for detection and enumeration of Campylobacter spp. Part 1: Detection method The European Standard EN ISO 10272-1:2006 has the status of a British Standard ICS 07.100.30 ? Licensed Copy: sheffiel
2、dun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS EN ISO 10272-1:2006 This British Standard was published under the authority of the Standards Policy and Strategy Committee on 3 February 2006 BSI 3 February 2006 ISBN 0 580 47687 1 National foreword This British S
3、tandard is the official English language version of EN ISO 10272-1:2006. It is identical with ISO 10272-1:2006. It supersedes BS 5763-17:1996 which is withdrawn. The UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology, which has the responsibility to: A list o
4、f organizations 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 Catalogue under the section entitled “International Standards
5、Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. Compliance with a British Standard does
6、 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 UK interests informed; monitor related international and European developmen
7、ts 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 v, a blank page, pages 1 to 16, an inside back cover and a back cover. The BSI copyright notice displayed
8、in this document indicates when the document was last issued. Amendments issued since publication Amd. No. DateComments Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10272-1 January
9、2006 ICS 07.100.30 English Version Microbiology of food and animal feeding stuffs - Horizontal method for detection and enumeration of Campylobacter spp. - Part 1: Detection method (ISO 10272-1:2006) Microbiologie des aliments - Mthode horizontale pour la recherche et le dnombrement des Campylobacte
10、r spp. - Partie 1: Mthode de recherche (ISO 10272-1:2006) Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum Nachweis und zur Zhlung von Campylobacter spp.- Teil 1: Nachweisverfahren (ISO 10272-1:2006) This European Standard was approved by CEN on 28 November 2005. CEN me
11、mbers 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 applica
12、tion to the Central Secretariat 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 Central Secretariat has the
13、 same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania
14、, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: rue de Stassart, 36 B-1050 Brussels 2006 CENAll rights of exploitation in any form and by any means reserved wo
15、rldwide for CEN national Members. Ref. No. EN ISO 10272-1:2006: E Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Foreword This document (EN ISO 10272-1:2006) has been prepared by Technical Committee ISO/TC 34 “Agricultural food products“ in
16、 collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods“, 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 July 2006, and con
17、flicting national standards shall be withdrawn at the latest by July 2006. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Fin
18、land, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. Endorsement notice The text of ISO 10272-1:2006 has been approved by CEN as EN ISO
19、 10272-1:2006 without any modifications. EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Reference number ISO 10272-1:2006(E) INTERNATIONAL STANDARD ISO 10272-1 First edition 2006-01-15 Microbiology of food and animal fee
20、ding stuffs Horizontal method for detection and enumeration of Campylobacter spp. Part 1: Detection method Microbiologie des aliments Mthode horizontale pour la recherche et le dnombrement de Campylobacter spp. Partie 1: Mthode de recherche EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun,
21、 na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ii Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI iii Contents Page Foreword iv Introduction v 1 Scope 1 2 Normative references1 3 Terms and definitions .1 4 Principle2 5 C
22、ulture media and reagents .2 6 Apparatus and glassware .3 7 Sampling.4 8 Preparation of test sample4 9 Procedure (see diagram in Annex A) 5 10 Expression of results 8 11 Test report8 Annex A (normative) Diagram of procedure 9 Annex B (normative) Composition and preparation of culture media and reage
23、nts.10 Bibliography16 EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 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 wo
24、rk 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-governm
25、ental, 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 2. The
26、main task of technical committees is to prepare International Standards. 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 75 % of the member bodies casting a vote.
27、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 rights. ISO 10272-1 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9, Microbio
28、logy. This first edition of ISO 10272-1, together with ISO/TS 10272-2:2006, cancels and replaces ISO 10272:1995, which has been technically revised. ISO 10272 consists of the following parts, under the general title Microbiology of food and animal feeding stuffs Horizontal method for detection and e
29、numeration of Campylobacter spp.: Part 1: Detection method Part 2: Colony-count technique (Technical Specification) EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI v Introduction Because of the large variety of food and f
30、eed products, this horizontal method may not be appropriate in every detail for certain products, and for some other products it may be necessary to use different methods. Nevertheless, it is hoped that in all cases every attempt will be made to apply this horizontal method as far as possible and th
31、at deviations from this will only be made if absolutely necessary for technical reasons. When this International Standard is next reviewed, account will be taken of all information then available regarding the extent to which this horizontal method has been followed and the reasons for deviations fr
32、om this in the case of particular products. The harmonization of test methods cannot be immediate and, for certain group of products, International Standards and/or national standards may already exist that do not comply with this horizontal method. It is hoped that when such standards are reviewed,
33、 they will be changed to comply with this International Standard so that eventually the only remaining departures from this horizontal method will be those necessary for well-established technical reasons. EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00
34、2006, Uncontrolled Copy, (c) BSI blank Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 1 Microbiology of food and animal feeding stuffs Horizontal method for detection and enumeration of Campylobacter spp. Part 1: Detection method 1 Scope Th
35、is part of ISO 10272 describes a horizontal method for the detection of Campylobacter spp. It is applicable to products intended for human consumption or for the feeding of animals, and to environmental samples in the area of food production and food handling, subject to the limitations stated in th
36、e Introduction. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 6887 (
37、all parts), Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and decimal dilutions for microbiological examination ISO 7218, Microbiology of food and animal feeding stuffs General rules for microbiological examinations ISO 8261, Milk and milk products Ge
38、neral guidance for the preparation of test samples, initial suspensions and decimal dilutions for microbiological examinations ISO/TS 11133-1, Microbiology of food and animal feeding stuffs Guidelines on preparation and production of culture media Part 1: General guidelines on quality assurance for
39、the preparation of culture media in the laboratory ISO/TS 11133-2:2003, Microbiology of food and animal feeding stuffs Guidelines on preparation and production of culture media Part 2: Practical guidelines on performance testing of culture media 3 Terms and definitions For the purposes of this docum
40、ent, the following terms and definitions apply. 3.1 Campylobacter microorganisms forming characteristic colonies on solid selective media when incubated microaerobically at 41,5 C but not at 25 C, and which possess the characteristic motility and biochemical and growth properties described when the
41、tests are conducted in accordance with this part of ISO 10272 EN ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 2 NOTE The most frequently encountered species are Campylobacter jejuni and Campylobacter coli. Other species h
42、ave, however, been described (Campylobacter lari, Campylobacter upsaliensis and some others). 3.2 detection of Campylobacter determination of the presence or absence of these microorganisms in a defined quantity of product, when the test is conducted in accordance with this part of ISO 10272 4 Princ
43、iple 4.1 General In general, the detection of Campylobacter requires the following stages (see Annex A for a diagram of the procedure). 4.2 Enrichment in selective liquid medium The test portion is inoculated into the liquid enrichment medium (Bolton broth) and homogenized. It is incubated in a micr
44、oaerobic atmosphere at 37 C for 4 h to 6 h and then at 41,5 C for 44 h 4 h. 4.3 Isolation and selection for confirmation From the cultures obtained in 4.2, two selective solid media are inoculated: modified charcoal cefoperazone deoxycholate agar (mCCD agar); any other solid selective medium based o
45、n a principle different from that of mCCD agar. They are then incubated at 41,5 C in a microaerobic atmosphere and inspected after 44 h 4 h to detect the presence of colonies presumed because of their characteristics to be Campylobacter. 4.4 Confirmation The colonies presumed to be Campylobacter are
46、 subcultured on the non-selective Columbia blood agar, then confirmed by means of microscopic examination and appropriate biochemical and growth tests. Optionally, the Campylobacter species are identified by specific biochemical tests and antibiotic sensitivity tests. 5 Culture media and reagents 5.
47、1 General For current laboratory practice, see ISO 7218, ISO/TS 11133-1 and ISO/TS 11133-2. NOTE Because of the large number of culture media and reagents and for the clarity of the text, their compositions and preparations are given in Annex B. 5.2 Liquid enrichment medium: Bolton broth See B.1. EN
48、 ISO 10272-1:2006 Licensed Copy: sheffieldun sheffieldun, na, Fri Nov 17 15:06:27 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 3 5.3 Selective plating medium: Modified charcoal cefoperazone deoxycholate agar (mCCD agar) See B.2. 5.4 Confirmation and identification media and reagents 5.4.1 Columbia blo
49、od agar See B.3. 5.4.2 Brucella broth See B.4. 5.4.3 Reagent for the detection of oxidase See B.5. 5.4.4 Hydrogen peroxide solution, 3 % (volume fraction) 5.4.5 Reagents for the detection of hydrolysis of hippurate See B.6. 5.4.6 Mueller Hinton blood agar See B.7. 5.4.7 Nalidixic acid (30 g) and cephalothin (30 g) discs 5.4.8 Indoxyl acetate discs See B.8. 6 Apparatus and glassware Usual microbiological laboratory equipment (see ISO 7218) and, in pa