Microbiology of the food chain - Methods for the detection of Anisakidae L3 larvae in fish and fishery products - Part 2: Artificial digestion method (ISO 23036-2:2021)

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, lightly processed fish products, such as marinated, salted or smoked, and it’s 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, which identification is made by morphological and/or molecular methods.

Mikrobiologie der Lebensmittelkette - Verfahren zum Nachweis von Anisakidae L3-Larven in Fisch und Fischereierzeugnissen - Teil 2: Verfahren der künstlichen Verdauung (ISO 23036-2:2021)

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 geräucherten Fischprodukten. Es ist auch als bestätigendes Verfahren bei einer Sichtprüfung von inneren Organen geeignet.
Das Verfahren der künstlichen Verdauung [4],[5],[6] ist anwendbar für die Quantifizierung parasitärer Infektionen durch Schätzung der Anzahl der in der Fischmuskulatur vorhandenen Parasiten. Bei der Anwendung mit frischem Fisch oder leicht verarbeiteten Fischprodukten (die vor der Verarbeitung nie eingefroren waren) kann zudem die Lebensfähigkeit der gegebenenfalls vorhandenen L3 Anisakidaelarven bestimmt werden.
Dieses Verfahren ist jedoch 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 les poissons et produits de la pêche - Partie 2: Méthode de digestion artificielle (ISO 23036-2:2021)

Mikrobiologija v prehranski verigi - Metode ugotavljanja prisotnosti ličink Anisakidae L3 v ribah in ribjih proizvodih – 2. del: Metoda umetne prebave (ISO 23036-2:2021)

General Information

Status
Published
Public Enquiry End Date
04-Feb-2020
Publication Date
29-Jun-2021
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
14-Jun-2021
Due Date
19-Aug-2021
Completion Date
30-Jun-2021

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SLOVENSKI STANDARD
SIST EN ISO 23036-2:2021
01-september-2021
Mikrobiologija v prehranski verigi - Metode ugotavljanja prisotnosti ličink
Anisakidae L3 v ribah in ribjih proizvodih – 2. del: Metoda umetne prebave (ISO
23036-2:2021)

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

and fishery products - Part 2: Artificial digestion method (ISO 23036-2:2021)
Mikrobiologie der Lebensmittelkette - Verfahren zum Nachweis von Anisakidae L3-

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

(ISO 23036-2:2021)
Microbiologie de la chaîne alimentaire - Méthodes de recherche des larves L3

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

artificielle (ISO 23036-2:2021)
Ta slovenski standard je istoveten z: EN ISO 23036-2:2021
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
SIST EN ISO 23036-2:2021 en

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

---------------------- Page: 1 ----------------------
SIST EN ISO 23036-2:2021
---------------------- Page: 2 ----------------------
SIST EN ISO 23036-2:2021
EN ISO 23036-2
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
2: Artificial digestion method (ISO 23036-2:2021)

Microbiologie de la chaîne alimentaire - Méthodes de Mikrobiologie der Lebensmittelkette - Verfahren zum

recherche des larves L3 d'Anisakidae dans les poissons Nachweis von Anisakidae L3-Larven in Fisch und

et produits de la pêche - Partie 2: Méthode de digestion Fischereierzeugnissen - Teil 2: Verfahren der

artificielle (ISO 23036-2:2021) künstlichen Verdauung (ISO 23036-2:2021)
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 23036-2:2021 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST EN ISO 23036-2:2021
EN ISO 23036-2:2021 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

---------------------- Page: 4 ----------------------
SIST EN ISO 23036-2:2021
EN ISO 23036-2:2021 (E)
European foreword

This document (EN ISO 23036-2:2021) 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 23036-2:2021 has been approved by CEN as EN ISO 23036-2:2021 without any

modification.
---------------------- Page: 5 ----------------------
SIST EN ISO 23036-2:2021
---------------------- Page: 6 ----------------------
SIST EN ISO 23036-2:2021
INTERNATIONAL ISO
STANDARD 23036-2
First edition
2021-04
Microbiology of the food chain —
Methods for the detection of
Anisakidae L3 larvae in fish and
fishery products —
Part 2:
Artificial digestion method
Microbiologie de la chaîne alimentaire — Méthodes de recherche des
larves L3 d'Anisakidae dans les poissons et produits de la pêche —
Partie 2: Méthode de digestion artificielle
Reference number
ISO 23036-2:2021(E)
ISO 2021
---------------------- Page: 7 ----------------------
SIST EN ISO 23036-2:2021
ISO 23036-2:2021(E)
COPYRIGHT PROTECTED DOCUMENT
© 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

below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2021 – All rights reserved
---------------------- Page: 8 ----------------------
SIST EN ISO 23036-2:2021
ISO 23036-2:2021(E)
Contents Page

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 V erification of findings ..................................................................................................................................................................... 2

5 Reagents ........................................................................................................................................................................................................................ 3

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

7 Sampling ........................................................................................................................................................................................................................ 4

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 ................................................................................................................... 5

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

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

9 Expression of the results .............................................................................................................................................................................. 5

10 Performance characteristics of the method ............................................................................................................................. 6

11 Test report ................................................................................................................................................................................................................... 6

12 Quality assurance ................................................................................................................................................................................................ 6

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

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

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

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

© ISO 2021 – All rights reserved iii
---------------------- Page: 9 ----------------------
SIST EN ISO 23036-2:2021
ISO 23036-2:2021(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

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 of the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO’s adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see www .iso .org/

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
---------------------- Page: 10 ----------------------
SIST EN ISO 23036-2:2021
ISO 23036-2:2021(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 anisakidosis, a disease that is not

only 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,

is also sometimes used). Worldwide, marine and wild anadromous fishes are intermediate hosts of

Anisakidae, whereas marine mammals are the definitive hosts.

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

[1]][2]

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

[3]
been validated and tested in multicentre collaborative studies (see Clause 10).
© ISO 2021 – All rights reserved v
---------------------- Page: 11 ----------------------
SIST EN ISO 23036-2:2021
---------------------- Page: 12 ----------------------
SIST EN ISO 23036-2:2021
INTERNATIONAL STANDARD ISO 23036-2:2021(E)
Microbiology of the food chain — Methods for the
detection of Anisakidae L3 larvae in fish and fishery
products —
Part 2:
Artificial digestion method
1 Scope

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 smoked. It is also suitable for visceral organs as a

confirmatory method for a visual inspection scheme.
[4][5][6]
The artificial digestion m
...

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-

1:2019)

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)

Mikrobiologie der Lebensmittelkette - Verfahren zum Nachweis von Anisakidae L3-

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

(ISO/DIS 23036-2:2019)
Microbiologie de la chaîne alimentaire - Méthodes de recherche des larves L3

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

artificielle (ISO/DIS 23036-2:2019)
Ta slovenski standard je istoveten z: prEN ISO 23036-2
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
oSIST prEN ISO 23036-2:2020 en

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

---------------------- Page: 1 ----------------------
oSIST prEN ISO 23036-2:2020
---------------------- Page: 2 ----------------------
oSIST prEN ISO 23036-2:2020
DRAFT INTERNATIONAL STANDARD
ISO/DIS 23036-2
ISO/TC 34/SC 9 Secretariat: AFNOR
Voting begins on: Voting terminates on:
2019-11-04 2020-01-27
Microbiology of the food chain — Methods for the
detection of Anisakidae L3 larvae in fish and fishery
products —
Part 2:
Artificial digestion method

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

poissons et produits de la pêche —
Partie 2: Méthode de digestion artificielle
ICS: 07.100.30
THIS DOCUMENT IS A DRAFT CIRCULATED
This document is circulated as received from the committee secretariat.
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
NOT BE REFERRED TO AS AN INTERNATIONAL
STANDARD UNTIL PUBLISHED AS SUCH.
IN ADDITION TO THEIR EVALUATION AS
ISO/CEN PARALLEL PROCESSING
BEING ACCEPTABLE FOR INDUSTRIAL,
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
Reference number
NATIONAL REGULATIONS.
ISO/DIS 23036-2:2019(E)
RECIPIENTS OF THIS DRAFT ARE INVITED
TO SUBMIT, WITH THEIR COMMENTS,
NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
PROVIDE SUPPORTING DOCUMENTATION. ISO 2019
---------------------- Page: 3 ----------------------
oSIST prEN ISO 23036-2:2020
ISO/DIS 23036-2:2019(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2019

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

below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved
---------------------- Page: 4 ----------------------
oSIST prEN ISO 23036-2:2020
ISO/DIS 23036-2:2019(E)
Contents Page

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

© ISO 2019 – All rights reserved iii
---------------------- Page: 5 ----------------------
oSIST prEN ISO 23036-2:2020
ISO/DIS 23036-2:2019(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 voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO's adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following

URL: www .iso .org/iso/foreword .html.

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

Microbiology.
A list of all parts in the ISO 23036 series can be found on the ISO website.
iv © ISO 2019 – All rights reserved
---------------------- Page: 6 ----------------------
oSIST prEN ISO 23036-2:2020
ISO/DIS 23036-2:2019(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 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

infected fish or cephalopods containing viable L3.

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

hosts of Anisakidae, whereas marine mammals are the definitive hosts.

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

been validated and tested in multicenter collaborative studies (see clause 11).

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

in this standard are demonstrated.
© ISO 2019 – All rights reserved v
---------------------- Page: 7 ----------------------
oSIST prEN ISO 23036-2:2020
---------------------- Page: 8 ----------------------
oSIST prEN ISO 23036-2:2020
DRAFT INTERNATIONAL STANDARD ISO/DIS 23036-2:2019(E)
Microbiology of the food chain — Methods for the
detection of Anisakidae L3 larvae in fish and fishery
products —
Part 2:
Artificial digestion method
1 Scope

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

made by morphological and/or molecular methods.
2 Normative references

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

microbiological examinations

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

3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

...

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