BS EN ISO 6887-6-2013 Microbiology of food and animal feed Preparation of test samples initial suspension and decimal dilutions for microbiological examination Specific rules for tr.pdf

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1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN ISO 6887-6:2013Microbiology of food andanimal feed Preparation oftest samples, initial suspensionand decimal dilutions formicrobiological examinationPart 6: Specific rules

2、for the preparation ofsamples taken at the primary productionstage (ISO 6887-6:2013)BS EN ISO 6887-6:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO6887-6:2013.The UK participation in its preparation was entrusted to TechnicalCommittee AW/9, Microbiolog

3、y.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2013. Published by BSI Standa

4、rdsLimited 2013ISBN 978 0 580 67812 7ICS 07.100.30Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 30 April 2013.Amendments issued since publicationDate Text affect

5、edEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 6887-6 March 2013 ICS 07.100.30 English Version Microbiology of food and animal feed - Preparation of test samples, initial suspension and decimal dilutions for microbiological examination - Part 6: Specific rules for the preparation of samp

6、les taken at the primary production stage (ISO 6887-6:2013) Microbiologie des aliments - Prparation des chantillons, de la suspension mre et des dilutions dcimales en vue de lexamen microbiologique - Partie 6: Rgles spcifiques pour la prparation des chantillons prlevs au stade de production primaire

7、 (ISO 6887-6:2013) Mikrobiologie von Lebensmitteln und Futtermitteln - Vorbereitung von Untersuchungsproben und Herstellung von Erstverdnnungen und von Dezimalverdnnungen fr mikrobiologische Untersuchungen - Teil 6: Spezifische Regeln fr die Vorbereitung von Proben aus der Primrproduktion (ISO 6887-

8、6:2013) This European Standard was approved by CEN on 16 February 2013. CEN members 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 bibliograp

9、hical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre 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 responsibili

10、ty of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic o

11、f Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATI

12、ON EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 6887-6:2013: EBS EN ISO 6887-6:2013EN ISO 6887-6:2013 (E) 3 Foreword This document (EN

13、ISO 6887-6:2013) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN, in collaboration with Technical Committee ISO/TC 34 “Food products“. This European Standard shall be given the status of a national standard, either by

14、publication of an identical text or by endorsement, at the latest by September 2013, and conflicting national standards shall be withdrawn at the latest by September 2013. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or

15、 CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech R

16、epublic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. B

17、S EN ISO 6887-6:2013ISO 6887-6:2013(E) ISO 2013 All rights reserved iiiContents PageForeword iv1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Diluents and disinfectants 25.1 Diluents for special purposes . 25.2 Disinfectants for use during laboratory examination . 36 Ap

18、paratus and glassware 37 Types of sample that can be sent to the laboratory . 38 Preparation of samples 38.1 General . 38.2 Storage . 49 Specific procedures 49.1 Procedures carried out on samples taken at farm . 49.2 Procedures carried out on samples taken from slaughterhouse . 69.3 Procedures for s

19、amples taken from poultry at hatchery or when transporting from hatchery to farm 710 Further decimal dilutions . 8Bibliography 9BS EN ISO 6887-6:2013ISO 6887-6:2013(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member b

20、odies). 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 established has the right to be represented on that committee. International organizations, governmental a

21、nd 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 standardization.International Standards are drafted in accordance with the rules given in the ISO/IEC Directives,

22、 Part 2.The 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 cast

23、ing a vote.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 6887-6 was prepared by the European Committee for Standardization (CEN) Technical Commi

24、ttee CEN/TC 275, Food analysis Horizontal methods, in collaboration with Technical Committee ISO/TC 34, Food products, Subcommittee SC 9, Microbiology, in accordance with the Agreement on technical cooperation between ISO and CEN (Vienna Agreement). ISO 6887 consists of the following parts, under th

25、e general title Microbiology of food and animal feed1) 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 Part 2: Specific rules for the preparation of meat a

26、nd meat products Part 3: Specific rules for the preparation of fish and fishery products Part 4: Specific rules for the preparation of products other than milk and milk products, meat and meat products, and fish and fishery products Part 5: Specific rules for the preparation of milk and milk product

27、s Part 6: Specific rules for the preparation of samples taken at the primary production stage1) It is intended that, upon revision, the main element of Parts 2 to 5 will be aligned with the main element of the title of Part 6 (i.e. “Microbiology of food and animal feed”).iv ISO 2013 All rights reser

28、vedBS EN ISO 6887-6:2013INTERNATIONAL STANDARD ISO 6887-6:2013(E)Microbiology of food and animal feed Preparation of test samples, initial suspension and decimal dilutions for microbiological examination Part 6: Specific rules for the preparation of samples taken at the primary production stage1 Sco

29、peThis part of ISO 6887 specifies rules for the preparation of samples taken at all stages from the farm to the slaughterhouse and their suspension for microbiological examination when the samples require different preparation from the methods described in ISO 6887-1. ISO 6887-1 defines the general

30、rules for the preparation of the initial suspension and decimal dilutions for microbiological examination.This part of ISO 6887 excludes the preparation of samples for both enumeration and detection test methods where preparation details are specified in the relevant International Standards.This par

31、t of ISO 6887 is applicable to various samples taken from the hatchery, the farm, from the vehicle or the animals during transportation, or from animals or their carcasses in the slaughterhouse, to indicate the microbiological status of the animals in relation to zoonotic agents. This part of ISO 68

32、87 does not apply to samples taken to assess the hygiene of meat. These are covered by ISO 6887-2.This part of ISO 6887 does not consider samples taken from the aquatic environment (marine or freshwater) at the primary production stage. These are covered by ISO 6887-3.2 Normative referencesThe follo

33、wing documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. 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, Micro

34、biology of food 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 dilutions1)ISO 7218:2007, Microbiology of food and animal feeding stuffs Gen

35、eral requirements and guidance for microbiological examinationsISO 13307, Microbiology of food and animal feed Primary production stage Sampling techniques3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1laboratory samplesample prepared for sendi

36、ng to the laboratory and intended for inspection or testing ISO 2013 All rights reserved 1BS EN ISO 6887-6:2013ISO 6887-6:2013(E)3.2test portionmeasured (volume or mass) representative sample taken from the laboratory sample (3.1) for use in the preparation of the initial suspension (3.4)3.3pooled s

37、amplecomposite sample taken from a number of different animals or composites of separate environmental samples3.4initial suspensionprimary dilution suspension, solution or emulsion obtained after a weighed or measured quantity of the product under examination (or of a test sample prepared from the p

38、roduct) has been mixed with, normally, a nine-fold quantity of diluent, allowing large particles, if present, to settleNote 1 to entry: Sometimes higher or lower initial dilutions are prepared, for example 1 in 5 or 1 in 100 (V1 V2).3.5further decimal dilutionssuspension or solution obtained by mixi

39、ng a measured volume of the initial suspension (3.4) with a nine-fold volume of diluent and by repeating this operation with each dilution prepared in this way, until a decimal dilution series, suitable for the inoculation of culture media, is obtained4 PrincipleAn initial suspension (3.4) is prepar

40、ed to obtain as uniform a distribution as possible of the microorganisms contained in the test sample, without reducing their viability.A suspension for pre-enrichment or enrichment is prepared in the same way, using the medium recommended by the method of analysis concerned, except in the special c

41、ases mentioned in each product subclause (section) of this part of ISO 6887.If necessary, further dilutions (3.5) are prepared in order to reduce the number of microorganisms per unit volume to allow, after incubation, observation of any growth (in the case of liquid media) or enumeration of colonie

42、s (in or on agar plates), as stated in each specific standard.In order to restrict, if required, the range of enumeration to a given interval, or if high numbers of microorganisms are foreseen, it is possible to inoculate only the appropriate dilutions (at least two successive dilutions) according t

43、o the calculation described in ISO 7218.5 Diluents and disinfectantsDiluents for general use are described in ISO 6887-1.5.1 Diluents for special purposes5.1.1 Neutralizing agentsNeutralizing liquid, prepared in accordance with ISO 13307, is used if needed at 10 % volume fraction in diluent. Neutral

44、izer is normally added when the sample is taken, before transportation to the laboratory.NOTE If a high level of formalin is expected in the samples, L-histidine (0.9 %) can be also added to the pre-enrichment broth.2 ISO 2013 All rights reservedBS EN ISO 6887-6:2013ISO 6887-6:2013(E)5.2 Disinfectan

45、ts for use during laboratory examinationDisinfectants are given in ISO 7218:2007, 6.2.4.6 Apparatus and glasswareUsual microbiological laboratory equipment for general use (see ISO 6887-1 and ISO 7218) and, in particular, the following:6.1 Homogenizers6.1.1 Peristaltic homogenizer (stomacher)6.1.2 R

46、otary homogenizer (blender)6.1.3 Vibrational mixer (pulsifier)6.2 Sterile hammer or plastic mallet6.3 Sterile sand, pestle and mortar6.4 Sterile forceps, scissors, scalpels, spatulas, spoons6.5 Sterile flasks or screw-cap bottles of appropriate capacities6.6 Sterile total delivery graduated pipettes

47、, pipette tips7 Types of sample that can be sent to the laboratoryThe samples shall be taken and transported in accordance with ISO 7218 and ISO 13307.The following list includes examples only. More detailed information on samples needed in different situations is given in Clause 9: samples taken at

48、 the farm: from the environment (e.g. swabs, litter, faeces, dust, water); from animals (e.g. swabs); samples taken at slaughter (e.g. rectal or caecal content, mesenteric lymph nodes); samples taken at the hatchery (e.g. hatcher basket liners, broken egg-shells); samples taken on vehicles, modules

49、and crates for animal transport (e.g. swabs).8 Preparation of samples8.1 GeneralAll preparations and manipulations should be carried out using good aseptic techniques and with sterile equipment to prevent microbial contamination of samples from all external sources (see ISO 7218). ISO 2013 All rights reserved 3BS EN ISO 6887-6:2013ISO 6887-6:2013(E)If the sample is transferred to another container and the whole laboratory sample is used, ensure that all of the sample is transferred (e.g. when transferring bootsocks fro