EN ISO 6579:2002/A1:2007
(Amendment)Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Salmonella spp. - Amendment 1: Annex D: Detection of Salmonella spp. in animal faeces and in environmental samples from the primary production stage (ISO 6579:2002/Amd 1:2007)
Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Salmonella spp. - Amendment 1: Annex D: Detection of Salmonella spp. in animal faeces and in environmental samples from the primary production stage (ISO 6579:2002/Amd 1:2007)
20060207 - ISO DIS running from 2005-09-20 to 2006-02-20. TC decision to opt for PQ followed by FV/VA ISO lead. ISO confirmation received that they will wait for PQ closure before embarking on parallel FDIS
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum Nachweis von Salmonella spp. - ÄNDERUNG 1: Anhang D: Nachweis von Salmonella spp. in Tierkot und in Umgebungsprobenproben aus der Primärpoduktion (ISO 6579:2002/Amd.1:2007)
Microbiologie des aliments - Méthode horizontale pour la recherche des Salmonella spp. - Amendement 1: Annexe D: Recherche des Salmonella spp. dans les matières fécales des animaux et dans des échantillons environnementaux au stade de la production primaire (ISO 6579:2002/Amd 1:2007)
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje prisotnosti vrst Salmonella - Dopolnilo 1: Dodatek D: Ugotavljanje vrst Salmonella v živalskih iztrebkih in v okoljskih vzorcih iz faz primarne proizvodnje (ISO 6579:2002/Amd 1:2007)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-oktober-2007
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje prisotnosti vrst
Salmonella - Dopolnilo 1: Dodatek D: Ugotavljanje vrst Salmonella v živalskih
iztrebkih in v okoljskih vzorcih iz faz primarne proizvodnje (ISO 6579:2002/Amd
1:2007)
Microbiology of food and animal feeding stuffs - Horizontal method for the detection of
Salmonella spp. - Amendment 1: Annex D: Detection of Salmonella spp. in animal faeces
and in environmental samples from the primary production stage (ISO 6579:2002/Amd
1:2007)
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum
Nachweis von Salmonella spp - Änderung 1 (ISO 6579:2002/Amd 1:2007)
Microbiologie des aliments - Méthode horizontale pour la recherche des Salmonella spp.
- Amendement 1: Annexe D: Recherche des Salmonella spp. dans les matieres fécales
des animaux et dans des échantillons environnementaux au stade de la production
primaire (ISO 6579:2002/Amd 1:2007)
Ta slovenski standard je istoveten z: EN ISO 6579:2002/A1:2007
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 6579:2002/A1
NORME EUROPÉENNE
EUROPÄISCHE NORM
July 2007
ICS 07.100.30
English Version
Microbiology of food and animal feeding stuffs - Horizontal
method for the detection of Salmonella spp. - Amendment 1:
Annex D: Detection of Salmonella spp. in animal faeces and in
environmental samples from the primary production stage (ISO
6579:2002/Amd 1:2007)
Microbiologie des aliments - Méthode horizontale pour la Mikrobiologie von Lebensmitteln und Futtermitteln -
recherche des Salmonella spp. - Amendement 1: Annexe Horizontales Verfahren zum Nachweis von Salmonella spp
D: Recherche des Salmonella spp. dans les matières - Änderung 1 (ISO 6579:2002/Amd 1:2007)
fécales des animaux et dans des échantillons
environnementaux au stade de la production primaire (ISO
6579:2002/Amd 1:2007)
This amendment A1 modifies the European Standard EN ISO 6579:2002; it was approved by CEN on 23 May 2007.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for inclusion of this
amendment into the relevant national standard without any alteration. Up-to-date lists and bibliographical references concerning such
national standards may be obtained on application to the CEN Management Centre or to any CEN member.
This amendment 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 CEN Management Centre has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, 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
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Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2007 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 6579:2002/A1:2007: E
worldwide for CEN national Members.
Foreword
This document (EN ISO 6579:2002/A1:2007) 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 January 2008, and conflicting national
standards shall be withdrawn at the latest by January 2008.
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, 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 6579:2002 has been approved by CEN as EN ISO 6579:2002/A1:2007 without
any modifications.
INTERNATIONAL ISO
STANDARD 6579
Fourth edition
2002-07-15
AMENDMENT 1
2007-07-15
Microbiology of food and animal feeding
stuffs — Horizontal method for the
detection of Salmonella spp.
AMENDMENT 1: Annex D: Detection of
Salmonella spp. in animal faeces and in
environmental samples from the primary
production stage
Microbiologie des aliments — Méthode horizontale pour la recherche
des Salmonella spp.
AMENDEMENT 1: Annexe D: Recherche des Salmonella spp. dans les
matières fécales des animaux et dans des échantillons
environnementaux au stade de la production primaire
Reference number
ISO 6579:2002/Amd.1:2007(E)
©
ISO 2007
ISO 6579:2002/Amd.1:2007(E)
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ii © ISO 2007 – All rights reserved
ISO 6579:2002/Amd.1:2007(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
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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
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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.
Amendment 1 to ISO 6579:2002 was prepared by Technical Committee ISO/TC 34, Food products,
Subcommittee SC 9, Microbiology.
ISO 6579:2002/Amd.1:2007(E)
Microbiology of food and animal feeding stuffs — Horizontal
method for the detection of Salmonella spp.
AMENDMENT 1: Annex D: Detection of Salmonella spp. in animal
faeces and in environmental samples from the primary production
stage
Page 1, Clause 2
Replace the introductory text as follows and add the two references.
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/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
Page 27, after Annex C
Add the following as Annex D.
ISO 6579:2002/Amd.1:2007(E)
Annex D
(normative)
Detection of Salmonella spp. in animal faeces and in environmental
samples from the primary production stage
D.1 Introduction
The method given in the main text of this International Standard is primarily intended for the isolation of
Salmonella spp. from food and feeding stuffs and is not always suitable for the detection of Salmonella spp.
from other matrices.
This annex is applicable to the detection of Salmonella spp. in
⎯ animal faeces (such as from poultry, pigs, cattle), and
⎯ environmental samples in the area of the primary production stage (such as dust).
The method in this annex is based upon Clause 9, with a different selective enrichment medium. Therefore,
where possible, reference will be made to Clause 9.
The selective enrichment medium as described in this annex (modified semi-solid Rappaport-Vassiliadis:
MSRV) is intended for the detection of motile Salmonellae and is not appropriate for the detection of
non-motile Salmonellae.
NOTE The non-motile Salmonella biovars of Salmonella Gallinarum (Salmonella Gallinarum biovar gallinarum and
Salmonella Gallinarum biovar pullorum do not seem to survive long in environmental samples and will therefore rarely be
detected in faecal or environmental (such as dust) samples (regardless of the method). The number of other non-motile
Salmonella serovars in faecal samples seems to be generally low. For example, in Reference [7] in which
circa 1 000 faecal samples of poultry layer flocks and circa 900 faecal samples of broiler flocks were analysed, less than
1 % of the total number of samples were positive in a selective broth and at the same time negative on MSRV (and likely
to be non-motile). Similar results were found in a Dutch study with circa 3 200 faecal samples of pigs (non-published data).
On the other hand, in the case of the study in Reference [7], up to almost 40 % of positive samples would not have been
detected (i.e. false negatives) if only a selective broth (in this case Rappaport Vassiliadis) had been used instead of a
semi-solid medium.
D.2 Principle
D.2.1 General
The detection of Salmonella in animal faeces and in samples of the primary production stage necessitates
four stages, as described in Clause 4.
D.2.2 Pre-enrichment in non-selective liquid medium
Buffered peptone water (BPW) is inoculated at ambient temperature with the test portion, then incubated
at 37 °C ± 1 °C for 18 h ± 2 h.
D.2.3 Enrichment on selective semi-solid medium
Modified semi-solid Rappaport-Vassiliadis (MSRV) agar plates are inoculated with the culture obtained
in D.2.2.
2 © ISO 2007 – All rights reserved
ISO 6579:2002/Amd.1:2007(E)
The MSRV is incubated at 41,5 °C ± 1 °C for 24 h ± 3 h. If a plate is negative after 24 h, it is incubated for a
further 24 h ± 3 h.
D.2.4 Selective plating and identification
From the culture obtained in D.2.3, two selective solid media are inoculated:
⎯ xylose lysine deoxycholate (XLD) agar;
⎯ any other solid selective medium complementary to XLD agar (see 4.4).
The XLD agar is incubated at 37 °C ± 1 °C and examined after 24 h ± 3 h.
The second selective medium is incubated in accordance with the manufacturer’s instructions.
D.2.5 Confirmation of identity
Colonies of presumptive Salmonella are subcultured, then plated-out as described in D.2.4, and their identity
is confirmed by means of appropriate biochemical and serological tests.
D.3 Culture media, reagents and sera
D.3.1 General
For current laboratory practice, see ISO 7218.
All media and reagents needed for this annex are described in Annex B, except for modified semi-solid
Rappaport-Vassiliadis (MSRV) medium, which is described in D.3.2. Alternatively, dehydrated complete media
or diluents may be used. Follow, in that respect, the manufacturer’s instructions.
NOTE The composition of MSRV, as described in Reference [8], contained 20 mg/l of novobiocin. However, from a
scientific point of view, 10 mg/l novobiocin is preferred. In studies performed at the CRL-Salmonella, more Salmonella-
positive results were found in pig faeces samples when tested with MSRV containing 10 mg/l than with MSRV containing
20 mg/l novobiocin (see Reference [9]). Furthermore, when testing different animal faeces (pigs, chicken, cattle) and
naturally contaminated dust, the migration zones on MSRV containing 10 mg/l novobiocin were (much) larger than on
MSRV containing 20 mg/l novobiocin (Reference [9]). The influence of novobiocin on bacterial motility was earlier
described in Reference [10].
For the preparation of the selective plating agar media (see B.4, XLD-agar), standard size Petri dishes may be
used (90 mm or 100 mm) instead of large size Petri dishes (140 mm).
ISO 6579:2002/Amd.1:2007(E)
D.3.2 Modified semi-solid Rappaport-Vassiliadis medium (MSRV)
D.3.2.1 Base medium
D.3.2.1.1 Composition
Enzymatic digest of animal and plant tissue 4,6 g
Acid hydrolys
...
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