1、BRITISH STANDARDBS EN ISO 10272-1:2006Microbiology of food and animal feeding stuffs Horizontal method for detection and enumeration of Campylobacter spp. Part 1: Detection methodThe European Standard EN ISO 10272-1:2006 has the status of a British StandardICS 07.100.30g49g50g3g38g50g51g60g44g49g42g
2、3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58BS EN ISO 10272-1:2006This British Standard was published under the authority of the Standards Policy and Strategy Committee on 3
3、February 2006 BSI 3 February 2006ISBN 0 580 47687 1National forewordThis British Standard 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 It supersedes BS 5763-17:1996 which is withdrawn.The UK participation in its preparation was entrusted to
4、Technical Committee AW/9, Microbiology, which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary.Cross-referencesThe British Standards which implement international or European publications referred to in this document may be
5、found in the BSI Catalogue under the section entitled “International Standards 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 re
6、sponsible for its correct application. Compliance with a British Standard does 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 kee
7、p UK interests informed; monitor related international and European developments and promulgate them in the UK.Summary of pagesThis 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
8、, an inside back cover and a back cover.The BSI copyright notice displayed in this document indicates when the document was last issued.Amendments issued since publicationAmd. No. Date CommentsEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 10272-1January 2006ICS 07.100.30English VersionMicrob
9、iology of food and animal feeding stuffs - Horizontalmethod for detection and enumeration of Campylobacter spp. -Part 1: Detection method (ISO 10272-1:2006)Microbiologie des aliments - Mthode horizontale pour larecherche et le dnombrement des Campylobacter spp. -Partie 1: Mthode de recherche (ISO 10
10、272-1:2006)Mikrobiologie von Lebensmitteln und Futtermitteln -Horizontales Verfahren zum Nachweis und zur Zhlung vonCampylobacter spp.- Teil 1: Nachweisverfahren (ISO10272-1:2006)This European Standard was approved by CEN on 28 November 2005.CEN members are bound to comply with the CEN/CENELEC Inter
11、nal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Central Secretariat or to any CEN member.T
12、his European Standard exists in three official versions (English, French, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the officialversions.CEN members are the
13、 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,Slovakia, Slovenia, Spain, Sweden, Switzerland and United
14、Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2006 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 10272-1:2006: E
15、Foreword This document (EN ISO 10272-1:2006) has been prepared by Technical Committee ISO/TC 34 “Agricultural food products“ in 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 s
16、tatus of a national standard, either by publication of an identical text or by endorsement, at the latest by July 2006, and conflicting national standards shall be withdrawn at the latest by July 2006. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the fol
17、lowing countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Sp
18、ain, Sweden, Switzerland and United Kingdom. Endorsement notice The text of ISO 10272-1:2006 has been approved by CEN as EN ISO 10272-1:2006 without any modifications. EN ISO 10272-1:2006Reference numberISO 10272-1:2006(E)INTERNATIONAL STANDARD ISO10272-1First edition2006-01-15Microbiology of food a
19、nd animal feeding 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 10272-1:2006ii iiiContents Page Forewor
20、d iv Introduction v 1 Scope 1 2 Normative references 1 3 Terms and definitions .1 4 Principle2 5 Culture 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 report 8 Annex A (normative
21、) Diagram of procedure 9 Annex B (normative) Composition and preparation of culture media and reagents .10 Bibliography 16 EN ISO 10272-1:2006iv Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of p
22、reparing 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-governmental,
23、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 main ta
24、sk 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. Attenti
25、on 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, Microbiology. T
26、his 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 enumerat
27、ion of Campylobacter spp.: Part 1: Detection method Part 2: Colony-count technique (Technical Specification) EN ISO 10272-1:2006vIntroduction Because of the large variety of food and feed products, this horizontal method may not be appropriate in every detail for certain products, and for some other
28、 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 that deviations from this will only be made if absolutely necessary for technical reasons. When this International Sta
29、ndard 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 from this in the case of particular products. The harmonization of test methods cannot be immediate and, for certain g
30、roup 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, they will be changed to comply with this International Standard so that eventually the only remaining departures fr
31、om this horizontal method will be those necessary for well-established technical reasons. EN ISO 10272-1:2006blank1Microbiology of food and animal feeding stuffs Horizontal method for detection and enumeration of Campylobacter spp. Part 1: Detection method 1 Scope This part of ISO 10272 describes a
32、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 the Introduction. 2 Normative refer
33、ences 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 (all parts), Microbiology of food
34、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 General guidance for the preparatio
35、n 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 the preparation of culture media
36、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 document, the following terms and defi
37、nitions 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 tests are conducted in accordance
38、 with this part of ISO 10272 EN ISO 10272-1:20062 NOTE The most frequently encountered species are Campylobacter jejuni and Campylobacter coli. Other species have, however, been described (Campylobacter lari, Campylobacter upsaliensis and some others). 3.2 detection of Campylobacter determination of
39、 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 Principle 4.1 General In general, the detection of Campylobacter requires the following stages (see Annex A for a diagram of the procedure). 4.2 E
40、nrichment in selective liquid medium The test portion is inoculated into the liquid enrichment medium (Bolton broth) and homogenized. It is incubated in a microaerobic 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 ob
41、tained in 4.2, two selective solid media are inoculated: modified charcoal cefoperazone deoxycholate agar (mCCD agar); any other solid selective medium based on 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 t
42、o detect the presence of colonies presumed because of their characteristics to be Campylobacter. 4.4 Confirmation The colonies presumed to be Campylobacter are subcultured on the non-selective Columbia blood agar, then confirmed by means of microscopic examination and appropriate biochemical and gro
43、wth tests. Optionally, the Campylobacter species are identified by specific biochemical tests and antibiotic sensitivity tests. 5 Culture media and reagents 5.1 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
44、 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 ISO 10272-1:200635.3 Selective plating medium: Modified charcoal cefoperazone deoxycholate agar (mCCD agar) See B.2. 5.4 Confirmation and id
45、entification media and reagents 5.4.1 Columbia blood 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
46、 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 particular, the following. 6.1 Apparatus for dry sterilization (oven) or wet sterilization (autocla
47、ve) See ISO 7218. 6.2 Oven, laminar flow cabinet or incubator, capable of operating between 37 C and 55 C. 6.3 Incubator, capable of operating at 41,5 C 1 C. 6.4 Water baths, capable of operating at 25 C 1 C and 37 C 1 C, or incubators capable of operating at 25 C 1 C and 37 C 1 C. 6.5 Water bath, c
48、apable of operating between 47 C and 50 C. 6.6 pH-meter, accurate to within 0,1 pH units at 25 C. EN ISO 10272-1:20064 6.7 Containers, in particular culture tubes of dimensions 18 mm 180 mm and 9 mm 180 mm, haemolysis tubes of dimensions 13 mm 75 mm, bottles with non-toxic metal closures and/or flas
49、ks of appropriate capacity with appropriate covers. 6.8 Petri dishes, in glass or plastic, with diameters 90 mm to 100 mm. 6.9 Total-delivery graduated pipettes, with a wide opening, and a nominal capacity of 1 ml and 10 ml, graduated in 0,1 ml divisions, and Pasteur pipettes. 6.10 Rubber teats, or any other safety system capable of being adapted to the graduated pipettes. 6.11 Sterile loops, of platinum/iridium, nickel/chromium or plastic, approximately 3 mm in diameter, and wires of the same
