oSIST prEN ISO 23036-2:2020

SLOVENSKI STANDARD
oSIST prEN ISO 23036-2:2020
01-februar-2020

Mikrobiologija v prehranski verigi - Metode ugotavljanja prisotnosti Anisakidae L3

larv v ribah in ribjih proizvodih - 2. del: Umetna digestivna metoda (ISO/DIS 23036-

Microbiology of the food chain - Methods for the detection of Anisakidae L3 larvae in fish

and fishery products - Part 2: Artificial digestion method (ISO/DIS 23036-2:2019)

Larven in Fisch und Fischereierzeugnissen - Teil 2: Verfahren der künstlichen Verdauung

d'Anisakidae dans les poissons et produits de la pêche - Partie 2: Méthode de digestion

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

Microbiologie de la chaîne alimentaire — Méthodes de recherche des larves L3 d'Anisakidae dans les

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Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Principle ........................................................................................................................................................................................................................ 2

4.1 General ........................................................................................................................................................................................................... 2

4.2 Sample size ................................................................................................................................................................................................. 2

4.3 Sample preparation ............................................................................................................................................................................ 2

4.4 Digestion of the sample ................................................................................................................................................................... 2

4.5 Filtration of the digest fluid ......................................................................................................................................................... 2

4.6 Verification of findings ..................................................................................................................................................................... 2

5 Reagents ........................................................................................................................................................................................................................ 2

6 Equipment and consumables .................................................................................................................................................................. 3

7 Sampling ........................................................................................................................................................................................................................ 3

8 Procedure..................................................................................................................................................................................................................... 4

8.1 Preparation of the sample. ............................................................................................................................................................ 4

8.2 Preparation of the digest fluid. .................................................................................................................................................. 4

8.3 Digestion of the sample in the glass beaker. .................................................................................................................. 4

8.4 Filtration of the digest fluid ......................................................................................................................................................... 5

8.5 Microscopic examination ............................................................................................................................................................... 5

9 Documentation ....................................................................................................................................................................................................... 5

10 Expression of the results. ............................................................................................................................................................................ 5

11 Performance characteristics of the method ............................................................................................................................. 5

Annex A (informative) Sample collection ........................................................................................................................................................ 7

Annex B (informative) Example of a laboratory worksheet for recording data when testing

fish fillets with artificial digestion method .............................................................................................................................. 8

Bibliography ................................................................................................................................................................................................................................ 9

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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,

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 L3 stage.

This stage is infective to fish and squid, and larvae are transferred between fishes through predation,

maintaining the L3 stage. Upon the host’s death, some larvae migrate from the abdominal cavity into

muscle tissues. Humans are incidental hosts and may be infected after ingesting raw or undercooked

Nematodes of the family Anisakidae, are the causative agents of human anisakidosis, a disease that not

only is a public health hazard affecting humans, but also represents an economic problem in fishery and

food safety (the term anisakiasis, designating the disease caused by members of the genus Anisakis, has

been used by some authors as well). Worldwide, marine and wild anadromous fishes are intermediate

Visual inspection procedures for the detection of Anisakidae larvae in fish are employed to minimize

the risk that contaminated fish will reach the consumer, thus preventing human anisakidosis.

The UV-press and the artificial digestion of the fish muscular tissue are the methods specifically

designed to detect nematode larvae in fish and to evaluate the infestation level of a batch, and have

Alternative methods can be used for analysis, provided their equivalence with the methods described

This part of ISO 23036 specifies a method that is applicable for the detection of Anisakidae L3 larvae

commonly found in marine and anadromous fishes. The method can be applied to fresh fish and/or

frozen fish, as well as lightly processed fish products, such as marinated, salted or smoked. It is also

suitable for visceral organs as confirmatory method for visual inspection scheme.

The artificial digestion method allows quantifying parasitic infections by estimating the number of

parasites in the fish musculature and, when applied to fresh fish or lightly processed fish products

(never frozen before processing), determining the viability of Anisakidae L3, which may be present.

This method doesn’t allow determining species or genotype of detected parasites. Final identification is

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 7218, Microbiology of food and animal feeding stuffs — General requirements and guidance for

CODEX STAN 244, 2004, Standard for salted Atlantic herring and salted sprat. CODEX STAN 244-2004.

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