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/FDAM 1:2026)

Mikrobiologie der Lebensmittelkette - Nachweis und Zählung von Cryptosporidium und Giardia in frischem grünem Blattgemüse und Beeren - Änderung 1: Verfahrensvalidierungsstudien und Leistungsmerkmale (ISO 18744:2016/FDAM 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/FDAM 1:2026)

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/FDAM 1:2026)

General Information

Status
Not Published
Publication Date
30-Aug-2026
Current Stage
5020 - Submission to Vote - Formal Approval
Start Date
02-Apr-2026
Completion Date
02-Apr-2026

Relations

Effective Date
12-Feb-2026
Effective Date
31-Jul-2024

Overview

EN ISO 18744:2016/FprA1 - Microbiology of the food chain - Detection of Cryptosporidium and Giardia in fresh leafy green vegetables and berry fruits - Amendment 1 focuses on the presence and detection methods for these significant protozoan pathogens within fresh produce. This amendment updates the original standard to refine the method's validation studies, incorporate performance characteristics, and clarify that the method is qualitative, reporting only presence or absence of these parasites in tested samples.

Ensuring food safety and public health, this standard reflects the latest guidance on accurate pathogen detection, supporting laboratories, food producers, and regulatory authorities in consistently assessing microbial contamination risks in fresh vegetables and berry fruits. By bringing together harmonized detection protocols and robust validation data, it underpins food microbiology best practices in the European and international context.

Key Topics

  • Target Organisms: Cryptosporidium oocysts and Giardia cysts, key protozoan pathogens in the food chain
  • Product Scope: Fresh leafy green vegetables (such as lettuce) and berry fruits (such as raspberries)
  • Method Update: Emphasis on the detection (not enumeration) of parasites and qualitative result reporting (presence/absence)
  • Validation Studies: Inclusion of interlaboratory trial data to support method robustness and reproducibility
  • Performance Characteristics: Standardizes evaluation criteria including sensitivity, specificity, limit of detection (LOD), and interlaboratory reproducibility. For example:
    • Low laboratory-to-laboratory variation for Cryptosporidium detection, indicating high reproducibility
    • Higher variability for Giardia detection, indicating lower reproducibility for this pathogen
  • Centrifugation Protocols: Updated technical notes clarify centrifugation options and their impact on recovery rates
  • Internal Controls: Method allows (but does not require) use of internal standard controls for test validation

Applications

This amendment to EN ISO 18744:2016 has significant practical applications in:

  • Food Testing Laboratories: Provides validated, internationally harmonized methods for detecting Cryptosporidium and Giardia, helping labs deliver reliable, comparable results across countries and sectors.
  • Food Industry Safety Programs: Supports food producers and processors in meeting regulatory and quality requirements for microbial safety in salad greens and berries, essential for both domestic markets and international trade.
  • Regulatory Oversight: Facilitates national food safety authorities and public health agencies in implementing consistent surveillance and outbreak investigation protocols for protozoan contamination in fresh produce.
  • Research and Risk Assessment: Offers standardized data and methods for stakeholders conducting food safety research or quantitative microbial risk assessments related to fresh produce.

By standardizing the approach for detecting these parasites, the standard helps reduce the risk of foodborne illness outbreaks and supports consumer protection globally.

Related Standards

  • ISO 16140 series: On validation of microbiological methods; referenced for method validation statistical approaches.
  • ISO 18744:2016: Original standard covering detection and enumeration (now updated to detection only) of Cryptosporidium and Giardia in leafy greens and berries.
  • Food Microbiology Standards:
    • EN ISO 11290 (Listeria in food products)
    • EN ISO 6579 (Salmonella in food products)
  • ICS Codes:
    • 07.100.30: Food microbiology
    • 67.080.01: Fruits, vegetables and derived products

EN ISO 18744:2016/FprA1 is essential for professionals seeking validated, robust protocols for parasite detection in fresh produce, ensuring compliance with international quality standards and strengthening the safety of the food supply chain.

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EN ISO 18744:2016/oprA1:2025

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Frequently Asked Questions

EN ISO 18744:2016/FprA1 is a draft published by the European Committee for Standardization (CEN). Its full title is "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/FDAM 1:2026)". This standard covers: 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/FDAM 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/FDAM 1:2026)

EN ISO 18744:2016/FprA1 is classified under the following ICS (International Classification for Standards) categories: 07.100.30 - Food microbiology. The ICS classification helps identify the subject area and facilitates finding related standards.

EN ISO 18744:2016/FprA1 has the following relationships with other standards: It is inter standard links to ISO 18744:2016/FDAmd 1, EN ISO 18744:2016. Understanding these relationships helps ensure you are using the most current and applicable version of the standard.

EN ISO 18744:2016/FprA1 is available in PDF format for immediate download after purchase. The document can be added to your cart and obtained through the secure checkout process. Digital delivery ensures instant access to the complete standard document.

Standards Content (Sample)


SLOVENSKI STANDARD
SIST EN ISO 18744:2016/oprA1:2025
01-maj-2025
Mikrobiologija v prehranski verigi - Ugotavljanje prisotnosti in števila
"Cryptosporidium" in "Giardia" v sveži zeleni listni zelenjavi in jagodičevju -
Dopolnilo A1: Študija validacije metode in značilnosti delovanja (ISO
18744:2016/DAmd 1:2025)
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/DAmd 1:2025)
Mikrobiologie der Lebensmittelkette - Nachweis und Zählung von Cryptosporidium und
Giardia in frischem grünem Blattgemüse und Beeren -Änderung 1 (ISO
18744:2016/DAmd 1:2025)
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/DAmd1:2025)
Ta slovenski standard je istoveten z: EN ISO 18744:2016/prA1:2025
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
SIST EN ISO 18744:2016/oprA1:2025 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

SIST EN ISO 18744:2016/oprA1:2025

SIST EN ISO 18744:2016/oprA1:2025
DRAFT
Amendment
ISO 18744:2016/
DAM 1
ISO/TC 34/SC 9
Microbiology of the food chain —
Secretariat: AFNOR
Detection and enumeration of
Cryptosporidium and Giardia in
Voting begins on:
2025-03-25
fresh leafy green vegetables and
Voting terminates on:
berry fruits
2025-06-17
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
ICS: 07.100.30
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IS THEREFORE SUBJECT TO CHANGE
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Reference number
ISO 18744:2016/DAM 1:2025(en)
SIST EN ISO 18744:2016/oprA1:2025
ISO 18744:2016/DAM 1:2025(en)
© ISO 2025
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Published in Switzerland
ii
SIST EN ISO 18744:2016/oprA1:2025
ISO 18744:2016/DAM 1:2025(en)
Foreword
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bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology.
A list of all parts in the ISO 18744 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
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Title
Delete the text:
and enumeration
Explanation note: WG6 agreed to remove the term enumeration in the title and text since, due to the nature of
the validation, the method cannot be considered quantitative but qualitative and the results should be reported
only as presence/absence of the parasite in the matrix. Any enumeration in the method refers to the number of
pre-labelled and enumerated non-viable Cryptosporidium oocysts and Giardia cysts that may be used as internal
controls. The use of internal controls is recommended not mandatory.
Replace the text with the following:
Microbiology of the food chain — Detection of Cryptosporidium and Giardia on fresh leafy green vegetables
and berry fruits
Contents
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iii
SIST EN ISO 18744:2016/oprA1:2025
ISO 18744:2016/DAM 1:2025(en)
Annex D (informative) Method validation studies and performance characteristics

Introduction
Line 13
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and for their enumeration
1  Scope
Line 1
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and enumeration
8.1
e)
NOTE 1
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Centrifugation at 2 500 x g can result in a very compact pellet. A lower speed of 1 100 x g for 10 min with no
braking has been reported to give at least equivalent recoveries of Cryptosporidium oocysts and Giardia cysts.
i)
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Centrifuge the eluate at 2 500 x g for 10 min with no braking.
NOTE 2
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Centrifugation at 2 500 x g can result in a very compact pellet. A lower speed of 1 100 x g for 10 min with no
braking has been reported to give at least equivalent recoveries of Cryptosporidium oocysts and Giardia cysts.

8.2
g)
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Centrifuge the eluate at 2 500 x g for 10 min with no braking.

NOTE 2
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Centrifugation at 2 500 x g can result in a very compact pellet. A lower speed of 1 100 x g for 10 min with no
braking has been reported to give at least equivalent recoveries of Cryptosporidium oocysts and Giardia cysts.

iv
SIST EN ISO 18744:2016/oprA1:2025
ISO 18744:2016/DAM 1:2025(en)
k)
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Centrifuge the eluate at 2 500 x g for 10 min with no braking.

NOTE 3
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Centrifugation at 2 500 x g can result in a very compact pellet. A lower speed of 1 100 x g for 10 min with no
braking has been reported to give at least equivalent recoveries of Cryptosporidium oocysts and Giardia cysts.

8.5.2.2
NOTE
Third sentence
Replace the text with the following:
Some Cryptosporidium species have diameters up to 8 μm (e.g., C. andersoni and C. muris, which could be
associated with infections in immunocompromised persons in rare cases).

10.1
First line
Replace the text with the following: The result should be given as the presence or absence of Cryptosporidium
oocysts and/or Giardia cysts detected per weight of fresh produce sample examined.

10.2
i)
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The presence or absence of Cryptosporidium oocysts and Giardia cysts, and any relevant details regarding
their identification
...

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