EN ISO 23036-1:2021

Microbiology of the food chain - Methods for the detection of Anisakidae L3 larvae in fish and fishery products - Part 1: UV-press method (ISO 23036-1:2021)

This document specifies a method for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method is applicable to fresh fish and/or frozen fish, as well as lightly processed fish products, such as marinated, salted or cold smoked.
This method is applicable to quantifying parasitic infections by estimating the number of parasites in the fish musculature.
This method does not apply to determining the species or genotype of detected parasites. Final identification is made by morphological and/or molecular methods.

Mikrobiologie der Lebensmittelkette - Verfahren zum Nachweis von Anisakidae L3-Larven in Fisch und Fischereierzeugnissen - Teil 1: UV-Pressverfahren (ISO 23036-1:2021)

Dieses Dokument legt ein Verfahren für den Nachweis von L3 Anisakidaelarven, die häufig in marinen und anadromen Fischen vorkommen, fest. Das Verfahren ist anwendbar bei frischem und/oder gefrorenem Fisch sowie bei leicht verarbeiteten Fischprodukten, wie etwa marinierten, gesalzenen oder kalt geräucherten Fischprodukten.
Dieses Verfahren ist anwendbar für die Quantifizierung parasitärer Infektionen durch Schätzung der Anzahl der in der Fischmuskulatur vorhandenen Parasiten.
Dieses Verfahren ist nicht für die Bestimmung der Art oder des Genotyps der nachgewiesenen Parasiten anwendbar. Eine endgültige Identifizierung erfolgt mittels morphologischer und/oder molekularer Verfahren.

Microbiologie de la chaîne alimentaire - Méthodes de recherche des larves L3 d’Anisakidae dans le poisson et les produits de la pêche - Partie 1: Méthode presse/UV (ISO 23036-1:2021)

Le présent document spécifie une méthode de détection des larves L3 d’Anisakidae qui se trouvent fréquemment dans les poissons marins et anadromes. La méthode s’applique aux poissons frais et/ou surgelés, ainsi qu’aux produits à base de poisson légèrement transformés, c’est-à-dire marinés, salés ou fumés à froid.
Cette méthode s’applique à la quantification des infestations parasitaires en estimant le nombre de parasites dans les tissus musculaires des poissons.
Cette méthode ne s’applique pas à la détermination des espèces ou du génotype des parasites détectés. Des méthodes morphologiques et/ou moléculaires permettent une identification définitive.

Mikrobiologija v prehranski verigi - Metode ugotavljanja prisotnosti ličink Anisakidae L3 v ribah in ribjih proizvodih – 1. del: Metoda UV-kompresije (ISO 23036-1:2021)

General Information

• Status: Published
• Publication Date: 11-May-2021
• Withdrawal Date: 29-Nov-2021
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 275/WG 6 - Microbiological contamination
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 12-May-2021
• Completion Date: 12-May-2021
• Directive: 2017/625 - Regulation (EU) 2017/625 of the European Parliament and of the Council of 15 March 2017 on official controls and other official activities performed to ensure the application of food and feed law, rules on animal health and welfare, plant health and plant protection products
• Directive: 882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
• Ref Project: SIST EN ISO 23036-1:2021 - Microbiology of the food chain - Methods for the detection of Anisakidae L3 larvae in fish and fishery products - Part 1: UV-press method (ISO 23036-1:2021)

Overview

EN ISO 23036-1:2021 - Microbiology of the food chain - Part 1: UV-press method - specifies a standardized laboratory procedure for the detection and quantification of Anisakidae L3 larvae in fish and fishery products. The method is applicable to fresh or frozen whole fish and fillets, and to lightly processed products (marinated, salted, cold-smoked). It estimates parasite burden in the fish musculature but does not determine species or genotype (final identification requires morphological and/or molecular methods).

Key topics and technical requirements

• Scope: Detection of third‑stage (L3) Anisakidae larvae (e.g., Anisakis, Contracaecum, Pseudoterranova) in marine and anadromous fish.
• Principle: Larvae fluoresce under UV light after the sample is pressed thin and frozen; fluorescence is due to lipofuscin pigments.
• Sample preparation: Fillets or fish sides are placed in clear plastic bags, pressed to a thin layer (approximately 1–2 mm), frozen, then thawed for inspection.
• Inspection: Visual screening under a 366 nm UV light reveals brightly fluorescent larvae as spots; infected areas are recorded to estimate parasite numbers in musculature.
• Applicability to large fish: Large fillets (e.g., salmon, cod, halibut) can be cut into smaller pieces and processed separately.
• Limitations: The method does not identify species/genotype - it is a detection/quantification technique. There are no internal quality controls embedded in the method; laboratories should follow general microbiology QA (see ISO 7218).
• Validation: The UV‑press and artificial digestion methods have been validated and evaluated in collaborative studies (performance characteristics are provided in the standard).

Applications and users

Practical uses and stakeholders:

• Seafood processors and processors’ quality control teams for routine screening of raw materials
• Food safety laboratories and accredited testing facilities performing parasite surveillance
• Regulatory agencies and inspection services enforcing public health controls for fishery products
• Research groups studying fish parasitology, prevalence and distribution of Anisakidae
• Importers/exporters and supply‑chain auditors requiring standardized detection methods

Benefits include consistent detection of Anisakidae larvae, support for food safety decision-making, and quantification of infection levels for risk assessment and batch acceptance.

Related standards

• ISO 7218 - General requirements and guidance for microbiological examinations (quality/QA)
• EN ISO 23036 series - other parts (e.g., artificial digestion methods) that complement the UV‑press approach

Keywords: ISO 23036-1:2021, EN ISO 23036-1, UV-press method, Anisakidae L3 detection, fish parasite screening, seafood safety, anisakidosis prevention.

Standards Content (Sample)

SLOVENSKI STANDARD
01-september-2021
Mikrobiologija v prehranski verigi - Metode ugotavljanja prisotnosti ličink
Anisakidae L3 v ribah in ribjih proizvodih – 1. del: Metoda UV-kompresije (ISO
23036-1:2021)
Microbiology of the food chain - Methods for the detection of Anisakidae L3 larvae in fish
and fishery products - Part 1: UV-press method (ISO 23036-1:2021)
Mikrobiologie der Lebensmittelkette - Verfahren zum Nachweis von Anisakidae L3-
Larven in Fisch und Fischereierzeugnissen - Teil 1: UV-Pressverfahren (ISO 23036-
1:2021)
Microbiologie de la chaîne alimentaire - Méthodes de recherche des larves L3
d’Anisakidae dans le poisson et les produits de la pêche - Partie 1: Méthode presse/UV
(ISO 23036-1:2021)
Ta slovenski standard je istoveten z: EN ISO 23036-1:2021
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 23036-1
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2021
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of the food chain - Methods for the detection
of Anisakidae L3 larvae in fish and fishery products - Part
1: UV-press method (ISO 23036-1:2021)
Microbiologie de la chaîne alimentaire - Méthodes de Mikrobiologie der Lebensmittelkette - Verfahren zum
recherche des larves L3 d'Anisakidae dans le poisson et Nachweis von Anisakidae L3-Larven in Fisch und
les produits de la pêche - Partie 1: Méthode presse/UV Fischereierzeugnissen - Teil 1: UV-Pressverfahren (ISO
(ISO 23036-1:2021) 23036-1:2021)
This European Standard was approved by CEN on 24 March 2021.

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, 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, Turkey 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
© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 23036-1:2021 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 23036-1:2021) 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 November 2021, and conflicting national standards
shall be withdrawn at the latest by November 2021.
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.
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, Turkey and the
United Kingdom.
Endorsement notice
The text of ISO 23036-1:2021 has been approved by CEN as EN ISO 23036-1:2021 without any
modification.
INTERNATIONAL ISO
STANDARD 23036-1
First edition
2021-04
Microbiology of the food chain —
Methods for the detection of
Anisakidae L3 larvae in fish and
fishery products —
Part 1:
UV-press method
Microbiologie de la chaîne alimentaire — Méthodes de recherche des
larves L3 d’Anisakidae dans le poisson et les produits de la pêche —
Partie 1: Méthode presse/UV
Reference number
ISO 23036-1:2021(E)
©
ISO 2021
ISO 23036-1:2021(E)
© ISO 2021
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
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Published in Switzerland
ii © ISO 2021 – All rights reserved

ISO 23036-1:2021(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Equipment and consumables . 2
6 Sampling . 2
7 Procedure. 2
7.1 Weighing the sample . 2
7.2 Preparation of the sample . 3
7.3 Pressing . 3
7.4 Freezing . 3
7.5 Thawing . 3
7.6 Visual inspection . 3
8 Expression of the results . 4
9 Performance characteristics of the method . 4
10 Test report . 4
11 Quality assurance . 4
Annex A (informative) Sample collection . 5
Annex B (informative) Findings after UV-press method . 6
Annex C (informative) Example of a laboratory worksheet for recording data when testing
fish fillets with the UV-press method . 8
Bibliography . 9
ISO 23036-1:2021(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
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ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
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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
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iso/ foreword .html.
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 275, Food analysis — Horizontal methods, in accordance with the Agreement on
technical cooperation between ISO and CEN (Vienna Agreement).
A list of all parts in the ISO 23036 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
complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2021 – All rights reserved

ISO 23036-1:2021(E)
Introduction
Nematodes of the Anisakidae family have a complex life cycle involving a high number of hosts. Adult
stages of Anisakidae reside in the stomach of marine mammals, where they are embedded in the mucosa.
Unembryonated eggs produced by adult females are released with the faeces of marine mammals and
become embryonated in seawater, where first-stage larvae (L1) develop in the eggs. The larvae moult
to become free-swimming second-stage larvae (L2) and, if ingested by crustaceans, mature into third-
stage larvae (L3). This stage is infective to fish and squid, and larvae are transferred between fishes
through predation, maintaining the L3 stage. Some larvae migrate from the abdominal cavity into
muscle tissues. Humans are incidental hosts and can be infected after ingesting raw or undercooked
infected fish or cephalopods containing viable L3.
Nematodes of the family Anisakidae are the causative agents of human
...