EN ISO 18744:2016
(Main)Microbiology of the food chain - Detection and enumeration of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits (ISO 18744:2016)
Microbiology of the food chain - Detection and enumeration of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits (ISO 18744:2016)
ISO 18744:2016 specifies a method that is applicable for the detection and enumeration of Cryptosporidium oocysts and Giardia cysts on or in food products that are described herein as fresh leafy green vegetables and berry fruits. With suitable controls, it may also be applicable for the examination of other fresh produce.
The microscopy descriptions are for Cryptosporidium spp. oocysts and Giardia duodenalis cysts of size ranges which include those species (Cryptosporidium) or assemblages (Giardia) known to be pathogenic to humans.
This method does not include any molecular analysis and therefore is not suitable for the determination of the species or genotypes/assemblages of Cryptosporidium oocysts and Giardia cysts. The method will detect all species and genotypes/assemblages that are known to be pathogenic for humans and also others that are not. For further identification, molecular typing assays are required. However, these cannot be reliably performed if process positive controls have been spiked into the samples, as the result of molecular typing assays will be obfuscated.
This method does not allow the determination of viability or infectivity of any Cryptosporidium oocysts and Giardia cysts which may be present.
Mikrobiologie der Lebensmittelkette - Nachweis und Zählung von Cryptosporidium und Giardia in frischem grünem Blattgemüse und Beeren (ISO 18744:2016)
Diese Internationale Norm legt ein Verfahren zum Nachweis und zur Zählung von Cryptosporidium-Oozysten und Giardia Zysten auf oder in Lebensmitteln, die hier als frisches grünes Blattgemüse und Beeren beschrieben werden, fest. Mit geeigneten Kontrollen ist es möglicherweise auch für die Untersuchung anderer Frischwaren geeignet.
Die Beschreibungen für die Mikroskopie sind für Oozysten von Cryptosporidium spp. und Zysten von Giardia duodenalis in den Größenbereichen, die die Arten (Cryptosporidium) oder Genogruppen (Giardia) einschließen, die als pathogen für den Menschen bekannt sind.
Dieses Verfahren schließt keine molekulare Analyse ein und ist deshalb nicht zur Bestimmung der Arten oder Genotypen/Genogruppen von Cryptosporidium Oozysten und Giardia Zysten geeignet. Mit dem Verfahren werden alle Arten und Genotypen/Genogruppen nachgewiesen, die als pathogen für den Menschen bekannt sind, und auch andere, die das nicht sind. Zur weiteren Identifizierung sind molekulare Typisierungstests erforderlich. Diese können jedoch nicht zuverlässig durchgeführt werden, wenn die Proben mit positiven Prozesskontrollen beimpft (en: spiked) wurden, da das Ergebnis von molekularen Typisierungstests verschleiert wird.
Dieses Verfahren ermöglicht keine Bestimmung der Lebensfähigkeit oder Infektiosität beliebiger möglicherweise vorhandener Cryptosporidium Oozysten und Giardia Zysten.
Microbiologie de la chaîne alimentaire - Recherche et dénombrement des Cryptosporidium et Giardia dans les légumes verts frais à feuilles et les fruits à baies (ISO 18744:2016)
ISO 18744:2016 spécifie une méthode applicable à la détection et au dénombrement des oocystes de Cryptosporidium et aux kystes de Giardia sur ou dans des produits alimentaires qui sont décrits ici comme étant des légumes verts frais à feuilles et des fruits à baies. Grâce à des contrôles appropriés, elle peut être également applicable à l'examen d'autres produits frais.
Les descriptions faites au microscope concernent les oocystes de Cryptosporidium spp. et les kystes de Giardia duodenalis dont la taille inclut les espèces (Cryptosporidium) ou assemblages (Giardia) connus pour être pathogènes pour l'homme.
Cette méthode n'inclut pas l'analyse moléculaire et n'est donc pas applicable à la détermination des espèces ou des génotypes/assemblages d'oocystes de Cryptosporidium et de kystes de Giardia. La méthode permettra de détecter toutes les espèces et tous les génotypes/assemblages connus pour être pathogènes pour l'homme ainsi que d'autres qui ne le sont pas. Pour mieux les identifier, des essais de typage moléculaire sont requis. Toutefois, ceux-ci ne peuvent pas être effectués de manière fiable si des contrôles de processus positifs ont été ajoutés aux échantillons, car le résultat des essais de typage moléculaire sera erroné.
Cette méthode ne permet pas de déterminer la viabilité ou l'infectivité des oocystes de Cryptosporidium et des kystes de Giardia qui peuvent être présents.
Mikrobiologija v prehranski verigi - Ugotavljanje prisotnosti in števila "Cryptosporidium" in "Giardia" v sveži zeleni listni zelenjavi in jagodičevju (ISO 18744:2016)
Ta mednarodni standard določa metodo, ki se uporablja za ugotavljanje prisotnosti in števila oocist »Cryptosporidium« in cist »Giardia« na ali v živilih, ki so v tem dokumentu opisana kot sveža zelena listna zelenjava in jagodičevje. Z ustreznim nadzorom se lahko uporablja tudi za preiskovanje drugih svežih proizvodov. Ta metoda ne omogoča ugotavljanja sposobnosti za življenje ali infektivnosti vseh oocist Cryptosporidium in cist Giardia, ki so lahko prisotne. Opisi mikroskopije se uporabljajo za oociste Cryptosporidium in ciste Giardia velikostnih obsegov, ki vključujejo vrste, za katere je znano, da so patogene za ljudi.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2016
0LNURELRORJLMDYSUHKUDQVNLYHULJL8JRWDYOMDQMHSULVRWQRVWLLQãWHYLOD
&U\SWRVSRULGLXPLQ*LDUGLDYVYHåL]HOHQLOLVWQL]HOHQMDYLLQMDJRGLþHYMX ,62
Microbiology of the food chain - Detection and enumeration of Cryptosporidium and
Giardia in fresh leafy green vegetables and berry fruits (ISO 18744:2016)
Mikrobiologie der Lebensmittelkette - Nachweis und Zählung von Cryptosporidium und
Giardia in frischem grünem Blattgemüse und Beeren (ISO 18744:2016)
Microbiologie de la châine alimentaire - Recherche et dénombrement des
Cryptosporidium et Giardia dans les légumes verts frais à feuilles et les fruits à baies
(ISO 18744:2016)
Ta slovenski standard je istoveten z: EN ISO 18744:2016
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.080.01 Sadje, zelenjava in njuni Fruits, vegetables and
proizvodi na splošno derived products in general
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN ISO 18744
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2016
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of the food chain - Detection and
enumeration of Cryptosporidium and Giardia in fresh leafy
green vegetables and berry fruits (ISO 18744:2016)
Microbiologie de la chaîne alimentaire - Recherche et Mikrobiologie der Lebensmittelkette - Nachweis und
dénombrement des Cryptosporidium et Giardia dans Zählung von Cryptosporidium und Giardia in frischem
les légumes verts frais à feuilles et les fruits à baies grünem Blattgemüse und Beeren (ISO 18744:2016)
(ISO 18744:2016)
This European Standard was approved by CEN on 11 March 2016.
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 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 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, 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
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2016 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 18744:2016 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
European foreword
This document (EN ISO 18744:2016) has been prepared by Technical Committee ISO/TC 34 “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 october 2016, and conflicting national standards shall
be withdrawn at the latest by october 2016.
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.
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, 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
The text of ISO 18744:2016 has been approved by CEN as EN ISO 18744:2016 without any modification.
INTERNATIONAL ISO
STANDARD 18744
First edition
2016-04-01
Microbiology of the food chain —
Detection and enumeration of
Cryptosporidium and Giardia in fresh
leafy green vegetables and berry fruits
Microbiologie de la chaîne alimentaire — Recherche et
dénombrement de Cryptosporidium et Giardia dans les légumes verts
frais à feuilles et les fruits à baies
Reference number
ISO 18744:2016(E)
©
ISO 2016
ISO 18744:2016(E)
© ISO 2016, Published in Switzerland
All rights reserved. Unless otherwise specified, 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
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2016 – All rights reserved
ISO 18744:2016(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Reagents . 2
6 Apparatus . 3
7 Sampling and transport . 4
7.1 Sampling . 4
7.2 Transport . 4
7.3 Receipt of samples . 4
7.4 Storage . 4
7.5 Preparation of test sample . 4
8 Procedure. 5
8.1 Removal of parasites from leafy green vegetables . 5
8.2 Removal of parasites from berry fruits . 6
8.3 Immunomagnetic separation (IMS) . 7
8.4 Sample staining . 7
8.5 Microscopy . 8
8.5.1 General comments . 8
8.5.2 Examination of sample preparations using epifluorescence microscopy . 9
8.5.3 Examination of sample preparations using DIC microscopy .11
9 Quality control procedures.12
9.1 General .12
9.2 Inclusion and interpretation of controls .12
9.3 Equipment cleaning .13
10 Reporting of results .13
10.1 Expression of results .13
10.2 Test report .13
Annex A (normative) Preparation of reagents .15
Annex B (normative) Preparation of positive process control suspensions .17
Annex C (informative) Percentage recoveries achieved by the methods .19
Bibliography .20
ISO 18744:2016(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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.