1、BRITISH STANDARD BS EN ISO 21567:2004 Microbiology of food and animal feeding stuffs Horizontal method for the detection of Shigella spp. The European Standard EN ISO 21567:2004 has the status of a British Standard ICS 07.100.30 BS EN ISO 21567:2004 This British Standard was published under the auth
2、ority of the Standards Policy and Strategy Committee on 22 November 2004 BSI 22 November 2004 ISBN 0 580 44803 7 National foreword This British Standard is the official English language version of EN ISO 21567:2004. It is identical with ISO 21567:2004. The UK participation in its preparation was ent
3、rusted to 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-references The British Standards which implement international or European publications referred to in this docu
4、ment may be 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.
5、 Users are responsible 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 ch
6、ange, 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 front cover, the EN ISO title page, the EN ISO foreword page, the ISO title page, pages ii to v, a blank
7、page, pages 1 to 26, 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 publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM ENISO21567 November2004 ICS07.100.30 E
8、nglishversion MicrobiologyoffoodandanimalfeedingstuffsHorizontal methodforthedetectionofShigellaspp.(ISO21567:2004) MicrobiologiedesalimentsMthodehorizontalepourla recherchedeShigellaspp.(ISO21567:2004) ThisEuropeanStandardwasapprovedbyCENon20October2004. CENmembersareboundtocomplywiththeCEN/CENELEC
9、InternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcernings uchnational standardsmaybeobtainedonapplicationtotheCentralSecretariatortoanyCENmember. ThisEuropeanStandardexistsinthree
10、officialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheCentralSecretariathasthesamestatusast heofficial versions. CENmembersarethenationalstandardsbodiesofAustria,Belgium,Cyprus,CzechRepublic,Denmark,Es
11、tonia,Finland,France, Germany,Greece,Hungary,Iceland,Ireland,Italy,Latvia,Lithuania,Luxembourg,Malta,Netherlands,Norway,Poland,Portugal, Slovakia, Slovenia,Spain,Sweden,SwitzerlandandUnitedKingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITEEFRNORMUNG Management
12、Centre:ruedeStassart,36B1050Brussels 2004CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.ENISO21567:2004:ENE ISO 12( 4002:765E) 2 Foreword This document (EN ISO 21567:2004) has been prepared by Technical Committee ISO/TC 34 “Agricultural food products
13、“ 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 status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 2005, and
14、conflicting national standards shall be withdrawn at the latest by May 2005. 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, F
15、inland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. Endorsement notice The text of ISO 21567:2004 has been approved by CEN as EN ISO 21567:20
16、04 without any modifications. ENISO21567:2004 Reference number ISO 21567:2004(E) OSI 4002INTERNATIONAL STANDARD ISO 21567 First edition 2004-11-01 Microbiology of food and animal feeding stuffs Horizontal method for the detection of Shigella spp. Microbiologie des aliments Mthode horizontale pour la
17、 recherche de Shigella spp. ENISO21567:2004IS:76512 O4002(E) DPlcsid Fremia ihTs PDF file may ctnoian emdebt dedyfepcaes. In ccaocnadrw eith Aebods licensilop gnic,y this file mairp eb ynted iv roweb detu slahl ton ide ebtlnu deess the typefaces whice era hml era deddebicsnede to i dnanstlaled t noe
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20、rm ttneC ehlar Secrteiraat ta the serddaig sleb nevwo. ISO 4002 All irthgs erse.devr lnUeto sswrehise specified, on trap fo this lbupictaion maeb y cudorperro de tuilizi den yna form ro na ybm ynae,s lecetrinoc ro mecinahcal, inclidung tohpcoiypodna gn micrfoilm, wittuoh repmissii non writign from i
21、etI rehSa Ot tsserdda eh ebolw or ISOs memreb i ydobn the cnuotrfo y ttseuqer ehe.r ISO cirypothg fofice saCe tsopale 65 eneG 1121-HC 02 av leT. 4 + 10 947 22 1 11 xaF0 947 22 14 + 9 74 E-mail coirypthgiso.o gr We bwww.is.o gro Pulbisdehi n Switlrez dnaii ISO 4002 Allr ithgsr esedevrENISO21567:2004I
22、S:76512 O4002(E) I SO 4002 All irthgs ersedevr iiiContents Page Foreword iv Introduction v 1 Scope 1 2 Normative references . 1 3 Terms and definitions. 2 4 Principle . 2 4.1 General. 2 4.2 Enrichment in selective liquid medium 2 4.3 Plating out and identification of colonies. 2 4.4 Biochemical and
23、serological confirmation. 2 5 Culture media, reagents and antisera. 2 6 Apparatus and glassware. 3 7 Sampling 3 8 Preparation of test sample. 3 9 Procedure (see diagram in Annex A) 4 9.1 Test portion . 4 9.2 Enrichment. 4 9.3 Plating out and colony selection. 4 9.4 Confirmation of colonies 4 9.5 Ser
24、ological confirmation 10 10 Expression of results 11 11 Test report 11 Annex A (normative) Diagram of test procedure 12 Annex B (normative) Composition and preparation of culture media and reagents 13 Annex C (normative) Description of Shigella colony morphology and colour on selective agars, for bo
25、th identification and quality control purposes 26 ENISO21567:2004IS:76512 O4002(E) iv I SO 4002 All irthgs ersedevrForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standar
26、ds 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, in liaison with ISO, also take
27、 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 task of technical committees is
28、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. Attention is drawn to the possibility
29、 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 21567 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9, Microbiology. ENISO21567:2004IS:76512 O4002(E)
30、I SO 4002 All irthgs ersedevr vIntroduction Because of the large variety of food and feed products, this horizontal method 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 justifi
31、ed technical reasons. Nevertheless, every attempt should be made to apply this horizontal method as far as possible. 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 t
32、he reasons for deviations from this method in the case of particular products. The harmonization of test methods cannot be immediate 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 th
33、at when such standards are reviewed 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. ENISO21567:2004blank 4002:76512OSINEINTENRATIONAL TSANDADR
34、IS:76512 O4002(E)I SO 4002 All irthgs ersedevr 1Microbiology of food and animal feeding stuffs Horizontal method for the detection of Shigella spp. WARNING In order to safeguard the health of laboratory personnel, it is essential that the whole of this method is only carried out by skilled personnel
35、 using good laboratory practices and preferably working in a containment facility. Relevant national Health and Safety Regulations relating to this organism shall be adhered to. Care shall be taken in the disposal of all infectious materials. 1 Scope This International Standard specifies a horizonta
36、l method for the detection of Shigella species. Subject to the limitations discussed in the Introduction, this International Standard is applicable to products intended for human consumption and the feeding of animals, and environmental samples in the area of food production and food handling. 2 Nor
37、mative 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 6887-1, Microbiology of foo
38、d and animal feeding stuffs Preparation of test samples, initial suspension and decimal dilutions for microbiological examination Part 1: General rules for the preparation of the initial suspension and decimal dilutions ISO 6887-2, Microbiology of food and animal feeding stuffs Preparation of test s
39、amples, initial suspension and decimal dilutions for microbiological examination Part 2: Specific rules for the preparation of meat and meat products ISO 6887-3, Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and decimal dilutions for microbiological e
40、xamination Part 3: Specific rules for the preparation of fish and fishery products ISO 6887-4, Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and decimal dilutions for microbiological examination Part 4: Specific rules for the preparation of products o
41、ther than milk and milk products, meat and meat products, and fish and fishery products ISO 7218:1996, Microbiology of food and animal feeding stuffs General rules for microbiological examinations ISO 8261, Milk and milk products General guidance for the preparation of test samples, initial suspensi
42、ons and decimal dilutions for microbiological examination 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 of culture media in the laboratory ISO/TS 11133-2, Microbiology of food a
43、nd animal feeding stuffs Guidelines on preparation and production of culture media Part 2: Practical guidelines on performance testing of culture media ENISO21567:2004IS:76512 O4002(E) 2 I SO 4002 All irthgs ersedevr3 Terms and definitions For the purposes of this document, the following terms and d
44、efinitions apply. 3.1 Shigella microorganisms which form colonies fitting the description of these species on the solid selective media used, and which display the biochemical and serological characteristics described when tests are carried out in accordance with this International Standard 3.2 dete
45、ction of Shigella spp. determination of the presence or absence of these microorganisms in a particular mass of product, when tests are carried out in accordance with this International Standard 4 Principle 4.1 General The detection of Shigella necessitates four successive stages (see Annex A). 4.2
46、Enrichment in selective liquid medium A test portion is inoculated into Shigella broth containing 0,5 g/ml of novobiocin, then incubated anaerobically at (41,5 1) C for 16 h to 20 h. 4.3 Plating out and identification of colonies From the enrichment culture obtained, three selective differential med
47、ia are inoculated: MacConkey agar with low selectivity; XLD agar with moderate selectivity; and Hektoen enteric agar with the greatest selectivity. All are incubated at 37 C for 20 h to 24 h. 4.4 Biochemical and serological confirmation Typical and suspect colonies are selected from each of the thre
48、e selective agars. The colonies are purified on nutrient agar, then biochemical and serological characterizations are carried out using the tests described. 5 Culture media, reagents and antisera For current laboratory practices, see ISO 7218, ISO/TS 11133-1 and ISO/TS 11133-2 for the preparation, production and performance testing of culture media. Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or demineralized water or water of equivalent purity.