SIST EN ISO 16654:2002/A2:2023

Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 - Amendment 2: Inclusion of performance testing of all culture media and reagents (ISO 16654:2001/Amd 2:2023)

Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren für den Nachweis von Escherichia coli O157 - Änderung 2: Aufnahme der Leistungsprüfung von Nährmedien und Reagenzien (ISO 16654:2001/Amd 2:2023)

No scope available

Microbiologie des aliments - Méthode horizontale pour la recherche des Escherichia coli O157 - Amendement 2: Inclusion d'essais de performance de tous les milieux de culture et réactifs (ISO 16654:2001/Amd 2:2023)

Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje Escherichia coli O157 - Dopolnilo A2: Vključitev preskušanja učinkovitosti vseh gojišč in reagentov (ISO 16654:2001/Amd 2:2023)

General Information

• Status: Published
• Public Enquiry End Date: 29-Apr-2022
• Publication Date: 11-Apr-2023
• ICS: 07.100.30 - Food microbiology
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 29-Mar-2023
• Due Date: 03-Jun-2023
• Completion Date: 12-Apr-2023
• Ref Project: EN ISO 16654:2001/A2:2023 - Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 - Amendment 2: Inclusion of performance testing of all culture media and reagents (ISO 16654:2001/Amd 2:2023)

Relations

• Amends: SIST EN ISO 16654:2002 - Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 (ISO 16654:2001)
• Effective Date: 08-Dec-2021

Overview

SIST EN ISO 16654:2002/A2:2023 is a key European Amendment to the international standard ISO 16654:2001, focusing on the microbiological detection of Escherichia coli O157 in food and animal feeding stuffs. This amendment introduces the inclusion of performance testing for all culture media and reagents used in the horizontal detection method of E. coli O157, enhancing the reliability and consistency of microbiological analyses.

Developed by ISO Technical Committee 34 in collaboration with CEN Technical Committee 463, this standard aligns with global food safety requirements. It supports laboratories and food safety professionals by ensuring standardized procedures for accurate, reproducible identification of E. coli O157, a critical pathogenic strain linked to serious foodborne illnesses.

Key Topics

• Horizontal Detection Method
  Defines a uniform test procedure applicable across various food matrices and animal feedingstuffs to detect E. coli O157.

• Performance Testing Requirements
  Establishes mandatory verification of culture media and reagents per ISO 11133, ensuring media productivity, selectivity, and specificity meet defined quality criteria before use.

• Culture Media and Reagents
  Includes detailed guidance on preparation, storage, shelf-life verification, and quality assurance of key media such as:

  • Modified Tryptone Soya Broth with novobiocin (mTSB + N)
  • Cefixime tellurite sorbitol MacConkey agar (CT-SMAC)
  • Nutrient agar and Tryptone/tryptophan media for confirmation tests

• Quality Control Strains
  Utilizes reference strains (e.g., E. coli O157:H7, Staphylococcus aureus) to validate media performance through growth characteristics and inhibition patterns.

• Normative References
  Integrates related ISO standards including ISO 11133 for culture media testing and ISO 7218 for general microbiological practices.

Applications

• Food Safety Testing
  Enables food manufacturers and testing laboratories to detect and monitor E. coli O157 contamination in raw materials, processed food products, and animal feeds.

• Regulatory Compliance
  Assists compliance with European food safety legislation and international guidelines by standardizing detection methods, thereby supporting public health protection against pathogenic E. coli outbreaks.

• Quality Assurance Programs
  Provides a framework for laboratories to implement rigorous quality control and validation procedures for culture media and reagents, ensuring consistent and reliable microbial test results.

• Research and Development
  Facilitates development of new detection platforms or improved culture media formulations by providing clear criteria for media performance testing.

Related Standards

• ISO 11133 – Microbiology of food, animal feed and water – Preparation, production, storage and performance testing of culture media. This standard underpins media quality assurance practices cited in SIST EN ISO 16654:2002/A2:2023.

• ISO 7218 – Microbiology of food and animal feeding stuffs – General requirements and guidance for microbiological examinations. It provides laboratory practice guidelines referenced as complementary to this standard.

• ISO 16649 Series – Standards related to the enumeration of E. coli and coliforms that complement detection efforts.

• National and International Food Safety Regulations – SIST EN ISO 16654:2002/A2:2023 supports compliance with EU food safety legislative frameworks and global trade standards.

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Keywords: E. coli O157 detection, food microbiology, animal feeding stuffs, culture media performance, microbiological testing standard, ISO 16654 amendment, food safety, microbiological quality assurance, horizontal method, CEN standards.

Standards Content (Sample)

SLOVENSKI STANDARD
01-maj-2023
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje Escherichia coli
O157 - Dopolnilo A2: Vključitev preskušanja učinkovitosti vseh gojišč in reagentov
(ISO 16654:2001/Amd 2:2023)
Microbiology of food and animal feeding stuffs - Horizontal method for the detection of
Escherichia coli O157 - Amendment 2: Inclusion of performance testing of all culture
media and reagents (ISO 16654:2001/Amd 2:2023)
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren für den
Nachweis von Escherichia coli O157 - Änderung 2: Aufnahme der Leistungsprüfung von
Nährmedien und Reagenzien (ISO 16654:2001/Amd 2:2023)
Microbiologie des aliments - Méthode horizontale pour la recherche des Escherichia coli
O157 - Amendement 2: Inclusion d'essais de performance de tous les milieux de culture
et réactifs (ISO 16654:2001/Amd 2:2023)
Ta slovenski standard je istoveten z: EN ISO 16654:2001/A2:2023
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 16654:2001/A2
EUROPEAN STANDARD
NORME EUROPÉENNE
February 2023
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of food and animal feeding stuffs - Horizontal
method for the detection of Escherichia coli O157 -
Amendment 2: Inclusion of performance testing of all
culture media and reagents (ISO 16654:2001/Amd
2:2023)
Microbiologie des aliments - Méthode horizontale pour Mikrobiologie von Lebensmitteln und Futtermitteln -
la recherche des Escherichia coli O157 - Amendement Horizontales Verfahren für den Nachweis von
2: Inclusion d'essais de performance de tous les Escherichia coli O157 - Änderung 2: Aufnahme der
milieux de culture et réactifs (ISO 16654:2001/Amd Leistungsprüfung von Nährmedien und Reagenzien
2:2023) (ISO 16654:2001/Amd 2:2023)
This amendment A2 modifies the European Standard EN ISO 16654:2001; it was approved by CEN on 23 December 2022.

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-CENELEC 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-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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2023 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16654:2001/A2:2023 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 16654:2001/A2:2023) has been prepared by Technical Committee ISO/TC 34
"Food products" in collaboration with Technical Committee CEN/TC 463 “Microbiology of the food
chain” the secretariat of which is held by AFNOR.
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 August 2023, and conflicting national standards shall
be withdrawn at the latest by August 2023.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
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, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 16654:2001/Amd 2:2023 has been approved by CEN as EN ISO 16654:2001/A2:2023
without any modification.
INTERNATIONAL ISO
STANDARD 16654
First edition
2001-05-01
AMENDMENT 2
2023-01
Microbiology of food and animal
feeding stuffs — Horizontal method
for the detection of Escherichia coli
O157
AMENDMENT 2: Inclusion of
performance testing of all culture media
and reagents
Microbiologie des aliments — Méthode horizontale pour la recherche
des Escherichia coli O157
AMENDEMENT 2: Inclusion d'essais de performance de tous les
milieux de culture et réactifs
Reference number
ISO 16654:2001/Amd.2:2023(E)
ISO 16654:2001/Amd.2:2023(E)
© ISO 2023
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
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