SIST EN ISO 10272-1:2006
(Main)Microbiology of food and animal feeding stuffs - Horizontal method for detection and enumeration of Campylobacter spp. - Part 1: Detection method (ISO 10272-1:2006)
Microbiology of food and animal feeding stuffs - Horizontal method for detection and enumeration of Campylobacter spp. - Part 1: Detection method (ISO 10272-1:2006)
This part of ISO 10272 describes a 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.
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum Nachweis und zur Zählung von Campylobacter spp.- Teil 1: Nachweisverfahren (ISO 10272-1:2006)
Dieser Teil von ISO 10272 legt ein horizontales Verfahren zum Nachweis von Campylobacter spp. fest.
Diese Internationale Norm gilt für Erzeugnisse, die für den menschlichen Verzehr oder als Futtermittel bestimmt sind, und für Umgebungsproben im Bereich der Herstellung und der Behandlung von Lebensmitteln, wobei die in der Einleitung angegebenen Einschränkungen gelten.
Microbiologie des aliments - Méthode horizontale pour la recherche et le dénombrement des Campylobacter spp. - Partie 1: Méthode de recherche (ISO 10272-1:2006)
L'ISO 10272-1:2006 décrit une méthode horizontale pour la recherche de Campylobacter spp.
Sous réserve de certaines restrictions (exposées dans l'introduction), elle est applicable aux produits destinés à la consommation humaine ou à l'alimentation des animaux, ainsi qu'aux échantillons environnementaux prélevés dans les secteurs de la production et de la manutention des aliments.
Mikrobiologija živil in krme – Horizontalna metoda za ugotavljanje prisotnosti in števila Campylobacter spp. – 1. del: Metoda za ugotavljanje prisotnosti (ISO 10272-1:2006)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 10272-1:2006
01-april-2006
Mikrobiologija živil in krme – Horizontalna metoda za ugotavljanje prisotnosti in
števila Campylobacter spp. – 1. del: Metoda za ugotavljanje prisotnosti (ISO 10272-
1:2006)
Microbiology of food and animal feeding stuffs - Horizontal method for detection and
enumeration of Campylobacter spp. - Part 1: Detection method (ISO 10272-1:2006)
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum
Nachweis und zur Zählung von Campylobacter spp.- Teil 1: Nachweisverfahren (ISO
10272-1:2006)
Microbiologie des aliments - Méthode horizontale pour la recherche et le dénombrement
des Campylobacter spp. - Partie 1: Méthode de recherche (ISO 10272-1:2006)
Ta slovenski standard je istoveten z: EN ISO 10272-1:2006
ICS:
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 10272-1:2006 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 10272-1:2006
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SIST EN ISO 10272-1:2006
EUROPEAN STANDARD
EN ISO 10272-1
NORME EUROPÉENNE
EUROPÄISCHE NORM
January 2006
ICS 07.100.30
English Version
Microbiology of food and animal feeding stuffs - Horizontal
method for detection and enumeration of Campylobacter spp. -
Part 1: Detection method (ISO 10272-1:2006)
Microbiologie des aliments - Méthode horizontale pour la Mikrobiologie von Lebensmitteln und Futtermitteln -
recherche et le dénombrement des Campylobacter spp. - Horizontales Verfahren zum Nachweis und zur Zählung von
Partie 1: Méthode de recherche (ISO 10272-1:2006) Campylobacter spp.- Teil 1: Nachweisverfahren (ISO
10272-1:2006)
This European Standard was approved by CEN on 28 November 2005.
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 bibliographical references concerning such national
standards may be obtained on application to the Central Secretariat 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 responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official
versions.
CEN members are the 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 Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2006 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10272-1:2006: E
worldwide for CEN national Members.
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SIST EN ISO 10272-1:2006
EN ISO 10272-1:2006 (E)
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 status 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 following 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, Spain, 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.
2
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SIST EN ISO 10272-1:2006
INTERNATIONAL ISO
STANDARD 10272-1
First edition
2006-01-15
Microbiology of food and animal
feeding stuffs — Horizontal method for
detection and enumeration of
Campylobacter spp. —
Part 1:
Detection method
Microbiologie des aliments — Méthode horizontale pour la recherche et
le dénombrement de Campylobacter spp. —
Partie 1: Méthode de recherche
Reference number
ISO 10272-1:2006(E)
©
ISO 2006
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
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ii © ISO 2006 – All rights reserved
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope .1
2 Normative references .1
3 Terms and definitions .1
4 Principle.2
5 Culture media and reagents .2
6 Apparatus and glassware .3
7 Sampling.4
8 Preparation of test sample.4
9 Procedure (see diagram in Annex A) .5
10 Expression of results .8
11 Test report .8
Annex A (normative) Diagram of procedure .9
Annex B (normative) Composition and preparation of culture media and reagents .10
Bibliography .16
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
Foreword
ISO (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
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 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 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 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.
This 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 enumeration of Campylobacter spp.:
⎯ Part 1: Detection method
⎯ Part 2: Colony-count technique (Technical Specification)
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
Introduction
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 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 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 the reasons for deviations from
this in the case of particular products. The harmonization of test methods cannot be immediate and, for certain
group 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 from this horizontal
method will be those necessary for well-established technical reasons.
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SIST EN ISO 10272-1:2006
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SIST EN ISO 10272-1:2006
INTERNATIONAL STANDARD ISO 10272-1:2006(E)
Microbiology 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 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 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 (all parts), Microbiology of food 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 preparation 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 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 definitions 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 with this part of ISO 10272
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
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 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 Enrichment 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 obtained 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 to 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 growth 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 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.
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
5.3 Selective plating medium: Modified charcoal cefoperazone deoxycholate agar (mCCD
agar)
See B.2.
5.4 Confirmation and identification 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 agar
See B.7.
5.4.7 Nalidixic acid (30 µg) and cephalothin (30 µg) 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 (autoclave)
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, capable of operating between 47 °C and 50 °C.
6.6 pH-meter, accurate to within 0,1 pH units at 25 °C.
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SIST EN ISO 10272-1:2006
ISO 10272-1:2006(E)
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 flasks 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 material, or a glass or plastic rod.
A nickel/chromium loop is not suitable for use in the oxidase test (see 9.4.6).
6.12 Forceps, fine, round-ended, of stainless steel.
6.13 Microscope, preferably with phase contrast (for observing the characteristic motility of Campylobacter).
6.14 Apparatus suitable for achieving a microaerobic atmosphere with oxygen content of 5 % ± 2 %,
carbon dioxide 10 % ± 3 %, optional hydrogen u 10 %, with the balance nitrogen. Appropriate gastight
containers are used to hold Petri dishes and/or flasks or bottles of about 350 ml capacity used for the
enrichment broth, e.g. bacteriological anaerobic jars.
NOTE 1 The appropriate microaerobic atmosphere can be obtained using commercially available gas-generating kits,
following precisely the manufacturer's instructions, particularly those relating to the volume of the jar and the capacity of
the gas-generating kit. Alternatively, the jar may be filled with an appropriate gas mixture prior to incubation.
NOTE 2 As an alternative to incubation in a microaerobic atmosphere, the enrichment broth can be incubated in screw-
capped bottles or flasks filled with enrichment broth, leaving a headspace of less than 2 cm, and tightly closing the caps.
7 Sampling
It is important that the laboratory receive a sample which is truly representative and has not been damaged or
changed during transport or storage.
Sampling is not part of the method specified in this part of ISO 10272. See the specific International Standard
dealing with the product concerned. If there is no specific International Standard dealing with sampling of the
product concerned, it is recommended that the parties concerned come to an agreement on this subject.
Since Campylobacter spp. are very sensitive to freezing but survive best at low temperatures, it is
recommended that samples to be tested should not be frozen, but stored at +3 °C ± 2 °C and subjected to
analysis as rapidly as possible. Also take care to prevent the samples from drying.
8 Preparation of test sample
Prepare the test sample in accordance with the specific International Standard dealing with the product
concerned. If there is no specific International Standard,
...
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