1、BRITISH STANDARDBS EN ISO 6888-1:1999IncorporatingAmendment No. 1 and Corrigendum No. 1Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of coagulase-positivestaphylococci(Staphylococcus aureusand other species) Part 1: Technique using Baird-Parker agar mediumThe E
2、uropean Standard EN ISO 6888-1:1999, with the incorporation of amendment A1:2003, has the status of a British StandardICS 07.100.30BS EN ISO 6888-1:1999This British Standard, havingbeen prepared under thedirection of the ConsumerProducts and Services SectorCommittee, was publishedunder the authority
3、 of theStandards Committee andcomes into effect on15 May 1999 BSI August 2006ISBN 0 580 32306 4National forewordThis British Standard is the English language version of EN ISO 6888-1:1999,including amendment A1:2003. It is identical with ISO 6888-1:1999, includingamendment 1:2003. Together with Part
4、 2, it supersedes BS 5763-7:1983 whichis withdrawn. This standard also supersedes BS 4285-3.10.1:1989 which iswithdrawn.The start and finish of text introduced or altered by amendment is indicated inthe text by tags . Tags indicating changes to CEN text carry the numberof the CEN amendment. For exam
5、ple, text altered by CEN amendment A1 isindicated by .The UK participation in its preparation was entrusted to Technical CommitteeAW/9, Microbiology, which has the responsibility to:A list of organizations represented on this committee can be obtained onrequest to its secretary.Cross-referencesThe B
6、ritish Standards which implement international or Europeanpublications referred to in this document may be found in the BSI Catalogueunder the section entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of BritishStandards Onl
7、ine.This publication does not purport to include all the necessary provisions of acontract. Users are responsible for its correct application.Compliance with a British Standard does not of itself confer immunityfrom legal obligations. aid enquirers to understand the text; present to the responsible
8、international/European committee anyenquiries on the interpretation, or proposals for change, and keep UK interests informed; monitor related international and European developments andpromulgate them in the UK.Summary of pagesThis document comprises a front cover, an inside front cover, the EN ISO
9、titlepage, the EN ISO foreword page, the ISO title page, pages ii to iv, pages 1 to 13and a back cover.The BSI copyright notice displayed in this document indicates when thedocument was last issued.Amendments issued since publicationAmd. No. Date Comments1459016271 August 2006 Addition of supersessi
10、on details tonational forewordCorrigendum No. 123 October 2003 See national forewordEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 6888-1February 1999+ A1July 2003ICS 07.100.30English versionMicrobiology of food and animal feeding stuffs Horizontal method for the enumeration of coagulase-posi
11、tive staphylococci (Staphylococcus aureusan d oth er sp ecies ) Pa rt 1: Tec hn iqu e u sin g Baird-Parker agar medium(includes amendment A1:2003)(ISO 6888-1:1999 + A1:2003)Microbiologie des aliments Mthode horizontale pour le dnombrement des staphylocoques coagulase positive (Staphylococcus aureus
12、et autres espces) Partie 1: Technique utilisant le milieu glos de Baird-Parker(inclut lamendement A1:2003)(ISO 6888-1:1999 + A1:2003)Mikrobiologie von Lebensmitteln und Futtermitteln Horizontales Verfahren fr die Zhlung von koagulase-positiven Staphylokokken (Staphylococcus aureus und andere Spezies
13、) Teil 1: Verfahren mit Baird Parker Agar(enthlt nderung A1:2003)(ISO 6888-1:1999 + A1:2003)This European Standard was approved by CEN on 15 February 1999.Amendment A1 was approved by CEN on 2 June 2003.CEN members are bound to comply with the CEN/CENELEC InternalRegulations which stipulate the cond
14、itions for giving this European Standardthe status of a national standard without any alteration. Up-to-date lists andbibliographical references concerning such national standards may be obtainedon application to the Central Secretariat or to any CEN member.This European Standard exists in three off
15、icial versions (English, French,German). A version in any other language made by translation under theresponsibility of a CEN member into its own language and notified to theCentral Secretariat has the same status as the official versions.CEN members are the national standards bodies of Austria, Bel
16、gium,Czech Republic, Denmark, Finland, France, Germany, Greece, Hungary,Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal,Slovakia, Spain, Sweden, Switzerland and United Kingdom.CENEuropean Committee for StandardizationComit Europen de NormalisationEuropisches Komitee fr Norm
17、ungCentral Secretariat: rue de Stassart 36, B-1050 Brussels 1999 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members.Ref. No. EN ISO 6888-1:1999 + A1:2003 EEN ISO 6888-1:1999 BSI 23 October 2003ForewordThe text of the International Standard from Te
18、chnical Committee ISO/TC 34, Agricultural food products, of the International Organization for Standardization (ISO) has been taken over as an European Standard by Technical Committee CEN/TC 275, Food analysis Horizontal methods, the Secretariat of which is held by DIN.This European Standard shall b
19、e given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by August 1999, and conflicting national standards shall be withdrawn at the latest by August 1999.According to the CEN/CENELEC Internal Regulations, the national standards organiza
20、tions of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom.Endorsement noticeThe tex
21、t of the International Standard ISO 6888-1:1999 has been approved by CEN as a European Standard without any modification.Foreword to amendment A1This document EN ISO 6888-1:1999/A1:2003 has been prepared by Technical Committee ISO/TC 34, Agricultural food products, in collaboration with Technical Co
22、mmittee 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 January 2004, and conflicting national standards shall
23、 be withdrawn at the latest by January 2004.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, Hungary, Iceland,
24、 Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and the United Kingdom.Endorsement noticeThe text of ISO 6888-1:1999/Amd 1 has been approved by CEN as EN ISO 6888-1:1999/A1:2003 without any modifications.EN ISO 6888-1:1999ii BSI 23 October 2003
25、ContentsPageForeword iii1Scope 12 Normative references 13 Terms and definitions 24 Principle 25 Diluent and culture media 26 Apparatus and glassware 57Sampling 58 Preparation of test sample 59 Procedure 510 Expression of results 711 Precision 912 Test report 10Annex A (informative) Results of the in
26、terlaboratory study 11Bibliography 13EN ISO 6888-1:1999 BSI 23 October 2003iiiForewordISO (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
27、 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, in liaison with ISO, also take part in the work. ISO collaborates cl
28、osely 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.Draft International Standards adopted by the technical committees are circulated
29、to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote.International Standard ISO 6888-1 was prepared by Technical Committee ISO/TC 34, Agricultural food products, Subcommittee SC 9, Microbiology.This first edi
30、tion of ISO 6888-1, together with ISO 6888-2, cancels and replaces ISO 6888:1983, which has been technically revised.ISO 6888 consists of the following parts, under the general title Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of coagulase-positive staphyloco
31、cci (Staphylococcus aureus and other species): Part 1: Technique using Baird-Parker agar medium; Part 2: Technique using rabbit plasma fibrinogen agar medium.ivblankEN ISO 6888-1:1999 BSI 23 October 200310 Introduction0.1 Because of the large variety of food and feed products, this horizontal method
32、 may not be appropriate in every detail for certain products. In this case, different methods, which are specific to these products, may be used if absolutely necessary for justified technical reasons. Nevertheless, every attempt should be made to apply this horizontal method as far as possible.When
33、 this part of ISO 6888 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 method in the case of particular products.The harmonization of test methods cannot be immed
34、iate and, for certain groups 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 part of ISO 6888 so that eventually the only remain
35、ing departures from this horizontal method will be those necessary for well-established technical reasons.0.2 ISO 6888 describes two horizontal methods (Part 1 and Part 2) for the enumeration of coagulase-positive staphylococci among which enterotoxinogenic strains are encountered. It is mainly conc
36、erned with Staphylococcus aureus, but also with S. intermedius and certain strains of S. hyicus.Both parts of ISO 6888 are given equivalent status. Nevertheless, it is recommended to use the procedure described in ISO 6888-2 (see reference 1) for the foods (such as cheeses made from raw milk and cer
37、tain raw meat products) likely to be contaminated by: staphylococci forming atypical colonies on a Baird-Parker agar medium; background flora which can obscure the colonies being sought.0.3 For the purposes of this part of ISO 6888, the confirmation of staphylococci is based on a positive coagulase
38、reaction, but it is recognized that some strains of Staphylococcus aureus give weakly positive coagulase reactions. These latter strains may be confused with other bacteria but they may be distinguished from such other bacteria by the use of additional tests not included in this part of ISO 6888, su
39、ch as the sensitivity to lysostaphin, the production of haemolysin, of thermostable nuclease and of acid from mannitol (see reference 2).1 ScopeThis part of ISO 6888 specifies a horizontal method for the enumeration of coagulase-positive staphylococci in products intended for human consumption or fe
40、eding of animals, by counting of colonies obtained on a solid medium (Baird-Parker medium) after aerobic incubation at 35 C or 37 C.2 Normative referencesThe following normative documents contain provisions which, through reference in this text, constitute provisions of this part of ISO 6888. For da
41、ted references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this part of ISO 6888 are encouraged to investigate the possibility of applying the most recent editions of the normative documents indicated below. For undated
42、references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain registers of currently valid International Standards.ISO 6887-1, Microbiology of food and animal feeding stuffs Rules for the preparation of the test sample, of initial suspension and of dec
43、imal dilutions for microbiological examination Part 1: General rules for the preparation of the initial suspension and of decimal dilutions.ISO 7218, Microbiology of food and animal feeding stuffs General rules for microbiological examination.EN ISO 6888-1:19992 BSI 23 October 20033 Terms and defini
44、tionsFor the purposes of this part of ISO 6888, the following terms and definitions apply.3.1coagulase-positive staphylococcibacteria which form typical and/or atypical colonies on the surface of a selective culture medium and which show a positive coagulase reaction when the test is performed follo
45、wing the method specified in this part of ISO 68883.2enumeration of the coagulase-positive staphylococcidetermination of the number of coagulase-positive staphylococci found per millilitre or per gram of sample when the test is carried out according to the method specified in this part of ISO 68884
46、Principle4.1 Inoculation of the surface of a solid selective culture medium, using duplicate plates, with a specified quantity of the test sample if the product is liquid, or with a specified quantity of the initial suspension in the case of other products.Inoculation, under the same conditions, usi
47、ng decimal dilutions of the test sample or of the initial suspension, with two plates per dilution.4.2 Aerobic incubation of the plates at 35 C or 37 C1)and examination after both 24 h and 48 h.4.3 Calculation of the number of coagulase-positive staphylococci per millilitre, or per gram, of sample f
48、rom the number of typical and/or atypical colonies obtained on plates at dilution levels chosen so as to give a significant result, and confirmed by a positive coagulase test result.5 Diluent and culture media5.1 GeneralFor current laboratory practice, see ISO 7218.5.2 DiluentSee ISO 6887-1 and the
49、specific standard dealing with the product to be examined.5.3 Baird-Parker agar medium2)NOTE Commercially available media may be used. In such cases, the manufacturers instructions should be followed carefully.5.3.1 Base medium5.3.1.1 Composition1)The temperature is agreed between the interested parties and is indicated in the test report.2)The agar medium is that of Baird-Parker (see reference 3) with the addition of sulfamezathine (see reference 4) if the presence of Proteus is
