EN ISO 18744:2016/A1:2026

• Status: Published
• Publication Date: 07-Jul-2026
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 463 - Microbiology of the food chain
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 08-Jul-2026
• Completion Date: 08-Jul-2026
• Ref Project: SIST EN ISO 18744:2016/A1:2026 - Microbiology of the food chain - Detection and enumeration of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits - Amendment 1: Method validation studies and performance characteristics (ISO 18744:2016/Amd 1:2026)

• Consolidated By: ISO 18744:2016/Amd 1:2026 - Microbiology of the food chain — Detection and enumeration of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits — Amendment 1: Method validation studies and performance characteristics
• Effective Date: 12-Feb-2026
• Amends: EN 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)
• Effective Date: 31-Jul-2024

Overview

EN ISO 18744:2016/A1:2026 is an amendment to the international standard developed by CEN and ISO focusing on the microbiology of the food chain, specifically the detection of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits. This amendment, known as Method Validation Studies and Performance Characteristics, delivers essential updates to the original ISO 18744:2016, clarifying the scope of the method as qualitative rather than quantitative and providing comprehensive validation data from interlaboratory studies.

Ensuring the accurate detection of Cryptosporidium oocysts and Giardia cysts in produce is crucial for safeguarding food safety and public health. This standard is especially relevant for food safety laboratories, regulatory bodies, and the food industry seeking to apply harmonized and validated microbiological testing protocols.

Key Topics

• Scope Clarification: The amendment reinforces that the method described is qualitative, i.e., for detection (presence/absence), not enumeration. That is, results are reported as whether the parasites are present or absent in the tested produce sample, rather than providing a count.
• Method Performance: Extensive interlaboratory studies were conducted on lettuce and raspberries (for Cryptosporidium) and lettuce (for Giardia) to determine specificity, sensitivity, and laboratory reproducibility.
  • Specificity and sensitivity levels are provided for different matrices and contamination levels.
  • Limit of Detection (LOD): Statistical methods were used to calculate LODs with confidence intervals, providing a benchmark for laboratories to assess method reliability.
• Centrifugation Procedure: Guidance is given on centrifugation parameters (2 500 x g for 10 minutes without braking), but also notes that a lower speed (1 100 x g for 10 minutes without braking) may yield similar recovery rates for oocysts and cysts.
• Reporting Requirements: Results must specify the presence or absence of Cryptosporidium oocysts and/or Giardia cysts for the tested sample, including details of any identification or verification performed.

Applications

EN ISO 18744:2016/A1:2026 is applicable to a broad range of stakeholders in the food chain, including:

• Food Safety Laboratories: Offers a standardized, validated detection procedure for identifying Cryptosporidium and Giardia in critical fresh produce categories. Laboratories can rely on this amendment for protocol harmonization and reliable performance parameters.
• Regulatory Bodies: Enables competent authorities and food regulators to enforce safety requirements for fresh leafy greens and berries more effectively, particularly regarding protozoan parasites.
• Food Producers and Processors: Supports risk assessment and compliance programs by providing a recognized test method for these pathogens, improving product safety and consumer confidence.
• Accreditation and Quality Control: Facilitates ISO/IEC 17025 accreditation by supplying robust, statistically validated method performance data, including information on interlaboratory reproducibility.
• Public Health Agencies: Enhances surveillance and outbreak investigation capabilities by standardizing the detection of these important waterborne and foodborne protozoa.

Related Standards

For organizations engaged in food microbiology and produce safety, the following standards are also relevant:

• ISO 16140-2: Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) microbiological methods.
• ISO 6887 series: Microbiology of food and animal feeding stuffs - Preparation of test samples, initial suspension, and decimal dilutions for microbiological examination.
• ISO 21527: Microbiology of food - Horizontal method for the enumeration of yeasts and moulds.
• EN ISO 18744:2016: The base standard for the detection of Cryptosporidium and Giardia in fresh produce; this amendment should be used in conjunction.
• Codex Alimentarius Guidelines: For microbiological criteria for foods.

By implementing EN ISO 18744:2016/A1:2026, organizations strengthen their food safety systems for detecting protozoan parasites in fresh produce, align with international best practices, and contribute to public health protection.

SLOVENSKI STANDARD
01-september-2026
Mikrobiologija v prehranski verigi - Ugotavljanje prisotnosti in števila
"Cryptosporidium" in "Giardia" v sveži zeleni listnati zelenjavi in jagodičju -
Dopolnilo A1: Študije validacije metode in značilnosti delovanja (ISO
18744:2016/Amd 1:2026)
Microbiology of the food chain - Detection and enumeration of Cryptosporidium and
Giardia in fresh leafy green vegetables and berry fruits - Amendment 1: Method
validation studies and performance characteristics (ISO 18744:2016/Amd 1:2026)
Mikrobiologie der Lebensmittelkette - Nachweis und Zählung von Cryptosporidium und
Giardia in frischem grünem Blattgemüse und Beeren - Änderung 1 (ISO
18744:2016/Amd 1:2026)
Microbiologie de la chaîne alimentaire - Recherche et dénombrement de
Cryptosporidium et Giardia dans les légumes verts frais à feuilles et les fruits à baies -
Amendement 1: Études de validation des méthodes et caractéristiques de performance
(ISO 18744:2016/Amd 1:2026)
Ta slovenski standard je istoveten z: EN ISO 18744:2016/A1:2026
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 18744:2016/A1
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2026
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 - Amendment 1: Method
validation studies and performance characteristics (ISO
18744:2016/Amd 1:2026)
Microbiologie de la chaîne alimentaire - Recherche et Mikrobiologie der Lebensmittelkette - Nachweis und
dénombrement de Cryptosporidium et Giardia dans les Zählung von Cryptosporidium und Giardia in frischem
légumes verts frais à feuilles et les fruits à baies - grünem Blattgemüse und Beeren - Änderung 1:
Amendement 1: Études de validation des méthodes et Verfahrensvalidierungsstudien und
caractéristiques de performance (ISO Leistungsmerkmale (ISO 18744:2016/Amd 1:2026)
18744:2016/Amd 1:2026)
This amendment A1 modifies the European Standard EN ISO 18744:2016; it was approved by CEN on 1 July 2026.

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
© 2026 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 18744:2016/A1:2026 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 18744:2016/A1:2026) 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 Amendment to the European Standard EN ISO 18744:2016 shall be given the status of a national
standard, either by publication of an identical text or by endorsement, at the latest by January 2027, and
conflicting national standards shall be withdrawn at the latest by January 2027.
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 18744:2016/Amd 1:2026 has been approved by CEN as EN ISO 18744:2016/A1:2026
without any modification.
International
Standard
ISO 18744
First edition
Microbiology of the food chain —
2016-04-01
Detection and enumeration of
Cryptosporidium and Giardia in
AMENDMENT 1
fresh leafy green vegetables and
2026-06
berry fruits
AMENDMENT 1: Method validation
studies and performance
characteristics
Microbiologie de la chaîne alimentaire — Recherche et
dénombrement de Cryptosporidium et Giardia dans les
légumes verts frais à feuilles et les fruits à baies
AMENDEMENT 1: Études de validation des méthodes et
caractéristiques de performance
Reference number
ISO 18744:2016/Amd.1:2026(en) © ISO 2026

ISO 18744:2016/Amd.1:2026(en)
© ISO 2026
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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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
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Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
ISO 18744:2016/Amd.1:2026(en)
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
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of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
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ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology, in collaboration with the European Committee for Standardization (CEN) Technical Committee
CEN/TC 463, Microbiology of the food chain, in accordance with the Agreement on technical cooperation
between ISO and CEN (Vienna Agreement).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iii
ISO 18744:2016/Amd.1:2026(en)
Microbiology of the food chain — Detection and
enumeration of Cryptosporidium and Giardia in fresh leafy
green vegetables and berry fruits
AMENDMENT 1: Method validation studies and performance
characteristics
Contents
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Annex D (informative) Method validation studies and performance characteristics
2. Normative references
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ISO 18744:2016/Amd.1:2026(en)
8.1, NOTE 1
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Centrifugation at 2 500g can result in a very compact pellet. A lower speed
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