DIN EN ISO 4833-1-2013 Microbiology of the food chain - Horizontal method for the enumeration of microorganisms - Part 1 Colony-count at 30 degrees C by the pour plate technique (I.pdf

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1、December 2013Translation by DIN-Sprachendienst.English price group 10No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).IC

2、S 07.100.30; 65.120!%*vJ“2078339www.din.deDDIN EN ISO 4833-1Microbiology of the food chain Horizontal method for the enumeration of microorganisms Part 1: Colony-count at 30 degrees C by the pour plate technique(ISO 4833-1:2013);English version EN ISO 4833-1:2013,English translation of DIN EN ISO 48

3、33-1:2013-12Mikrobiologie der Lebensmittelkette Horizontales Verfahren zur Zhlung von Mikroorganismen Teil 1: Koloniezhlung bei 30 C mittels Gussplattenverfahren (ISO 4833-1:2013);Englische Fassung EN ISO 4833-1:2013,Englische bersetzung von DIN EN ISO 4833-1:2013-12Microbiologie de la chane aliment

4、aire Mthode horizontale pour le dnombrement des micro-organismes Partie 1: Comptage des colonies 30 degrs C par la technique densemencement enprofondeur (ISO 4833-1:2013);Version anglaise EN ISO 4833-1:2013,Traduction anglaise de DIN EN ISO 4833-1:2013-12Together withDIN EN ISO 4833-2:2013-12,supers

5、edesDIN EN ISO 4833:2003-06www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 16 pages11.13 DIN EN ISO 4833-1:2013-12 2 A comma is used as the decimal marker. National foreword This document (EN ISO 4833-1:2013) has been prepared by Techni

6、cal Committee ISO/TC 34 “Food products” (Secretariat: AFNOR, France/ABNT, Brazil), Subcommittee SC 9 “Microbiology” (Secretariat: AFNOR, France) in collaboration with Technical Committee CEN/TC 275 “Food analysis Horizontal methods” (Secretariat: DIN, Germany). Based on the results of parallel votin

7、g, ISO 4833-1:2013 has been adopted as a European Standard. The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-01-06 AA Mikrobiologische Lebensmit

8、teluntersuchung einschlielich Schnellverfahren. ISO/TC 34/SC 9 decided to revise and subdivide EN ISO 4833 into two parts. EN ISO 4833 Part 1 specifies a horizontal method for the enumeration of microorganisms at 30 C by the pour plate technique and supersedes EN ISO 4833:2003. EN ISO 4833 Part 2 sp

9、ecifies a horizontal method for the enumeration of microorganisms at 30 C by the surface plating technique. DIN EN ISO 4833 consists of the following parts, under the general title Microbiology of the food chain Horizontal method for the enumeration of microorganisms: Part 1: Colony count at 30 C by

10、 the pour plate technique Part 2: Colony count at 30 C by the surface plating technique The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 6887 (all parts) DIN EN ISO 6887 (all parts) ISO 7218 DIN EN ISO 7218 ISO 11133 DIN EN ISO 11133 Ame

11、ndments This standard differs from DIN EN ISO 4833:2003-06 as follows: a) Clause 1 “Scope” has been rendered more precise and specifies in detail the application of the pour plate technique; b) Subclause 5.3.3 “Performance testing of the culture medium” has been added. Previous editions DIN 10192-1:

12、 1981-01, 1984-04 DIN EN ISO 4833: 2003-06 DIN EN ISO 4833-1:2013-12 3 National Annex NA (informative) Bibliography DIN EN ISO 6887 (all parts), Microbiology of food and animal feeding stuffs Preparation of the test samples, of initial suspension and of decimal dilutions for microbiological examinat

13、ion DIN EN ISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations DIN EN ISO 11133, Microbiology of food, animal feed and water Preparation, production, storage and performance testing of culture media DIN EN ISO 4833-1:2013-12 4 T

14、his page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 4833-1 September 2013 ICS 07.100.30 Supersedes EN ISO 4833:2003English Version Microbiology of the food chain - Horizontal method for the enumeration of microorganisms - Part 1: Colony count at 30 degrees C by t

15、he pour plate technique (ISO 4833-1:2013) Microbiologie de la chane alimentaire - Mthode horizontale pour le dnombrement des micro-organismes -Partie 1: Comptage des colonies 30 degrs C par la technique densemencement en profondeur(ISO 4833-1:2013) Mikrobiologie der Lebensmittelkette - Horizontales

16、Verfahren zur Zhlung von Mikroorganismen -Teil 1: Koloniezhlung bei 30 C mittels Gussplattenverfahren (ISO 4833-1:2013) This European Standard was approved by CEN on 26 July 2013. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this

17、 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 application to the CEN-CENELEC Management Centre or to any CEN member. This European Standard exists in three official ve

18、rsions (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 CEN-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austri

19、a, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, 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,

20、 Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC 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 n

21、ational Members. Ref. No. EN ISO 4833-1:2013: EContents Page Foreword . 3 1 Scope . 4 2 Normative references. 4 3 Terms and definitions 4 4 Principle 5 5 Culture media and diluents . 5 5.1 General 5 5.2 Diluents . 5 5.3 Agar medium: plate count agar (PCA) . 5 5.4 Overlay medium (if necessary; see 9.

22、2.7) . 6 6 Apparatus 7 7 Sampling . 7 8 Preparation of test sample 7 9 Procedure 7 9.1 Test portion, initial suspension and dilutions 7 9.2 Inoculation and incubation . 7 9.3 Counting of colonies 8 10 Expression of results . 8 10.1 Method of calculation 8 10.2 Precision . 8 10.3 Interpretation of te

23、st results . 9 11 Test report . 10 Annex A (informative) Use of the critical difference for the interpretation of results. 11 Bibliography 12 2DIN EN ISO 4833-1:2013-12EN ISO 4833-1:2013 (E) Foreword This document (EN ISO 4833-1:2013) has been prepared by Technical Committee ISO/TC 34 Food products

24、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 March 2014, and c

25、onflicting national standards shall be withdrawn at the latest by March 2014. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. This doc

26、ument supersedes EN ISO 4833:2003, together with EN ISO 4833-2:2013. 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 Republic, Denmark,

27、 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. Endorsement notice

28、 The text of ISO 4833-1:2013 has been approved by CEN as EN ISO 4833-1:2013 without any modification. “”3DIN EN ISO 4833-1:2013-12EN ISO 4833-1:2013 (E)1 ScopeThis part of ISO 4833 specifies a horizontal method for enumeration of microorganisms that are able to grow and form colonies in a solid medi

29、um after aerobic incubation at 30 C. The method is applicable to:a) products intended for human consumption and for animal feed;b) environmental samples in the area of food and feed production and handling.This part of ISO 4833 is applicable to:1) products that require a reliable count when a low li

30、mit of detection is specified (below 102/g or 102/ml for liquid samples or below 103/g for solid samples);2) products expected to contain spreading colonies that obscure colonies of other organisms, e.g. milk and milk products likely to contain spreading Bacillus spp.The applicability of this part o

31、f ISO 4833 to the examination of certain fermented food and animal feeds is limited and other media or incubation conditions can be more appropriate. However, this method can be applied to such products even though it is possible that the predominant microorganisms in those products are not detected

32、 effectively.For some matrices, the method specified in this part of ISO 4833 can give different results to those obtained using the method specified in ISO 4833-2.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable fo

33、r 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 (all parts), Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and d

34、ecimal dilutions for microbiological examinationISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinationsISO 11133, Microbiology of food, animal feed and water Preparation, production, storage and performance testing of culture media3

35、 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.4DIN EN ISO 4833-1:2013-12EN ISO 4833-1:2013 (E) 3.1microorganismentity of microscopic size, encompassing bacteria, fungi, protozoa and virusesSOURCE: ISO/TS 11139:2006,32.26Note 1 to entry: For the pu

36、rposes of this part of ISO 4833, microorganisms are bacteria, yeasts and moulds that are able to produce colonies under the conditions specified in this part of ISO 4833.4 PrincipleA specified quantity of the liquid test sample, or a specified quantity of an initial suspension in the case of other p

37、roducts, is dispensed into an empty Petri dish and mixed with a specified molten agar culture medium to form a poured plate.Other plates are prepared under the same conditions using decimal dilutions of the test sample or of the initial suspension.The plates are incubated under aerobic conditions at

38、 30 C for 72 h.The number of microorganisms per gram or per millilitre of the test sample is calculated from the number of colonies obtained in the plates containing fewer than 300 colonies.5 Culture media and diluents5.1 GeneralFollow ISO 11133 for preparation, production and performance testing of

39、 culture media.5.2 DiluentsUse the diluent(s) specified in ISO 6887 for the product concerned or the specific International Standard dealing with the product under examination.5.3 Agar medium: plate count agar (PCA)5.3.1 CompositionEnzymatic digestion of casein 5,0 gYeast extract 2,5 gGlucose, anhyd

40、rous (C6H12O6) 1,0 gAgara9 g to 18 gWater 1 000 mlaDepending on the gel strength of the agar.When dairy products are examined, add skimmed milk powder at 1,0 g/l of the culture medium. The skimmed milk powder shall be free from inhibitory substances.5.3.2 PreparationDissolve the components or the de

41、hydrated complete medium in the water, by heating if necessary. Mix thoroughly and leave to stand for several minutes.5DIN EN ISO 4833-1:2013-12EN ISO 4833-1:2013 (E)Adjust the pH (6.4), if necessary, so that after sterilization it is 7,0 0,2 at 25 C.Dispense the medium into tubes, flasks or bottles

42、 (6.8) of suitable capacity. Sterilize in an autoclave (6.1) at 121 C for 15 min.lf the medium is to be used immediately, cool it to 44 C to 47 C in a water bath (6.3) before use. If not, store it in the dark at a temperature of (5 3) C for no longer than 3 months, under conditions which do not allo

43、w any change in its composition and properties.Before beginning the microbiological examination, completely melt the medium, then cool it to 44 C to 47 C in a water bath (6.3) before use. See ISO 11133.Use the molten agar as soon as possible; it should not be retained for more than 4 h.5.3.3 Perform

44、ance testing of the culture medium5.3.3.1 GeneralPlate count agar is a non-selective medium, used in this part of ISO 4833 as a pour plate. Productivity shall be tested according to ISO 11133.5.3.3.2 ProductivityIncubation (30 1) C for (72 3) h Control strains Escherichia coli WDCM 00013 or WDCM 000

45、12aWorld Data Centre for Micro-organisms (WDCM)Bacillus subtilis subsp. spizizenii WDCM 00003aStaphylococcus aureus WDCM 00032 or WDCM 00034Reference medium Tryptone soya agarControl method QuantitativeCriterion Productivity ratio (PR) 0,7aThe strains to be used as a minimum by the user laboratory.

46、See Reference 7 for information on culture collection strain numbers and contact details.5.4 Overlay medium (if necessary; see 9.2.7)5.4.1 CompositionAgara12 g to 18 gWater 1 000 mlaDepending on the gel strength of the agar.5.4.2 PreparationAdd the agar to the water and heat to boiling, stirring fre

47、quently until the agar is completely dissolved, or steam for about 30 min.Adjust the pH (6.4), if necessary, so that after sterilization it is 7,0 0,2 at 25 C.Dispense the medium into tubes, flasks or bottles (6.8) of appropriate capacity.Sterilize in an autoclave at 121 C for 15 min.6DIN EN ISO 483

48、3-1:2013-12EN ISO 4833-1:2013 (E) lf the medium is to be used immediately, cool it to 44 C to 47 C in a water bath (6.3) before use. If not, store it in the dark at a temperature of (5 3) C for no longer than 3 months, under conditions which do not allow any change in its composition and properties.

49、Before beginning the microbiological examination, completely melt the medium then cool it to 44 C to 47 C in a water bath (6.3) before use. See ISO 11133.6 ApparatusDisposable apparatus is an acceptable alternative to re-usable glassware and plastic if it has suitable specifications.Usual microbiological laboratory equipment (see ISO 7218) and in particular the following.6.1 Oven for dry sterilization or autocla