Microbiology of the food chain - Preparation of test samples, initial suspension and decimal dilutions for microbiological examination - Part 3: Specific rules for the preparation of fish and fishery products (ISO 6887-3:2017)

ISO 6887-3:2017 specifies rules for the preparation of fish and fishery product samples and their suspension for microbiological examination when the samples require a different preparation from the methods described in ISO 6887‑1. ISO 6887‑1 defines the general rules for the preparation of the initial suspension and dilutions for microbiological examination.
ISO 6887-3:2017 includes special procedures for sampling raw molluscs, tunicates and echinoderms from primary production areas.
NOTE 1       Sampling of raw molluscs, tunicates and echinoderms from primary production areas is included in this document, rather than ISO 13307, which specifies rules for sampling from the terrestrial primary production stage.
ISO 6887-3:2017 excludes preparation of samples for both enumeration and detection test methods where preparation details are specified in the relevant International Standards (e.g. ISO/TS 15216‑1 and ISO/TS 15216‑2 for determination of hepatitis A virus and norovirus in food using real-time RT-PCR).
ISO 6887-3:2017 is intended to be used in conjunction with ISO 6887‑1. It is applicable to the following raw, processed or frozen fish and shellfish and their products (see Annex A for classification of major taxa):
a)    Raw fishery products, molluscs, tunicates and echinoderms including:
-      whole fish or fillets, with or without skin and heads, and gutted;
-      crustaceans, whole or shelled;
-      cephalopods;
-      bivalve molluscs;
-      gastropods;
-      tunicates and echinoderms.
b)    Processed products including:
-      smoked fish, whole or prepared fillets, with or without skin;
-      cooked or partially cooked, whole or shelled crustaceans, molluscs, tunicates and echinoderms;
-      cooked or partially cooked fish and fish-based multi-component products.
c)    Raw or cooked frozen fish, crustaceans, molluscs and others, in blocks or otherwise, including:
-      fish, fish fillets and pieces;
-      whole and shelled crustacean (e.g. flaked crab, prawns), molluscs, tunicates and echinoderms.
NOTE 2       The purpose of examinations performed on these samples can be either hygiene testing or quality control. However, the sampling techniques described in this document relate mainly to hygiene testing (on muscle tissues).

Mikrobiologie der Lebensmittelkette - Vorbereitung von Untersuchungsproben und Herstellung von Erstverdünnungen und von Dezimalverdünnungen für mikrobiologische Untersuchungen - Teil 3: Spezifische Regeln für die Vorbereitung von Fisch und Fischerzeugnissen (ISO 6887-3:2017)

Dieses Dokument legt Regeln für die Vorbereitung von Proben von Fisch und Fischerzeugnissen und deren Verdünnungen für mikrobiologische Untersuchungen fest, wenn die Proben eine Vorbereitung erfordern, die sich von den in ISO 6887 1 beschriebenen Verfahren unterscheidet. ISO 6887 1 legt die allgemeinen Regeln für die Herstellung der Erstverdünnung und der Verdünnungen für mikrobiologische Untersuchungen fest.
Dieses Dokument enthält spezielle Verfahren zur Probenahme von rohen Weichtieren, Manteltieren und Stachelhäutern aus Bereichen der Primärproduktion.
ANMERKUNG 1   Die Probenahme von rohen Weichtieren, Manteltieren und Stachelhäutern aus Bereichen der Primärproduktion ist in diesem Dokument statt in ISO 13307 enthalten, in der spezielle Regeln für die Probenahme in der Primärproduktion an Land festgelegt sind.
Dieses Dokument schließt die Vorbereitung von Proben für Prüfverfahren sowohl zur Zählung als auch zum Nachweis aus, bei denen die Einzelheiten zur Vorbereitung in den betreffenden Internationalen Normen festgelegt sind (z. B. ISO/TS 21872 1 und ISO/TS 21872 2 zum Nachweis von potentiell enteropathogenen Vibrio spp., ISO/TS 15216-1 und ISO/TS 15216-2 zur Bestimmung von Hepatitis A-Virus und Norovirus in Lebensmitteln mittels Real-time-RT-PCR).
Dieses Dokument ist vorgesehen um in Verbindung mit ISO 6887 1 angewendet zu werden. Er gilt für folgende rohe, verarbeitete oder gefrorene Fische und Schalentiere und die aus diesen hergestellten Produkten (hinsichtlich der Klassifikation der wichtigsten taxonomischen Gruppen (Taxa) siehe Anhang A):
a)   Rohe Fischerzeugnisse, Weichtiere, Manteltiere und Stachelhäuter, einschließlich:
   Fisch, ganz oder als Filet, mit oder ohne Haut und Köpfen sowie ausgenommen;
   Krebstiere im Ganzen oder geschält;
   Kopffüßler;
   Muscheln;
   Gastropoden;
   Manteltiere und Stachelhäuter.
b)   Verarbeitete Produkte, einschließlich:
   geräucherter Fisch, ganz oder als Filet, mit oder ohne Haut;
   gekochte oder teilweise gekochte Krebstiere, Weichtiere, Manteltiere und Stachelhäuter, ganz oder geschält;
   gekochter oder teilweise gekochter Fisch und Mehrkomponentenprodukte auf der Grundlage von Fisch.
c)   Rohe oder gekochte gefrorene Fische, Krebstiere, Weichtiere und andere, in Blöcken oder anderer Form, einschließlich:
   Fisch, Fischfilet und Fischstücke;
   ganze und geschälte Krebstiere (z. B. zerlegte Krabben, Garnelen), Weichtiere, Manteltiere und Stachelhäuter.
ANMERKUNG 2   Der Zweck der an diesen Proben durchgeführten Untersuchungen kann entweder eine Hygieneuntersuchung oder Qualitätskontrolle sein. Die in diesem Dokument beschriebenen Probenahmeverfahren beziehen sich jedoch hauptsächlich auf eine Hygieneuntersuchung (an Muskelgewebe).

Microbiologie de la chaîne alimentaire - Préparation des échantillons, de la suspension mère et des dilutions décimales en vue de l'examen microbiologique - Partie 3: Règles spécifiques pour la préparation des produits de la pêche (ISO 6887-3:2017)

ISO 6887-3:2017 spécifie des règles pour la préparation des échantillons de produits de la pêche et leur mise en suspension en vue de l'examen microbiologique, lorsque ces échantillons nécessitent une préparation différente des méthodes décrites dans l'ISO 6887‑1. L'ISO 6887‑1 définit des règles générales pour la préparation de la suspension mère et des dilutions en vue de l'examen microbiologique.
ISO 6887-3:2017 comprend des modes opératoires spécifiques pour le prélèvement de mollusques crus, tuniciers et échinodermes de zones de production primaire.
NOTE 1       Le prélèvement de mollusques crus, tuniciers et échinodermes de zones de production primaire est inclus dans le présent document plutôt que dans l'ISO 13307 qui spécifie des règles de prélèvement au stade de production primaire terrestre.
ISO 6887-3:2017 exclut la préparation d'échantillons pour les méthodes d'essai de dénombrement et de recherche où les détails de préparation sont spécifiés dans les Normes internationales concernées (par exemple, ISO/TS 15216‑1 and ISO/TS 15216‑2 pour la détermination du virus de l'hépatite A et du norovirus dans les aliments par RT-PCR en temps réel).
ISO 6887-3:2017 est destiné à être utilisé en lien avec l'ISO 6887‑1. Il est applicable aux poissons, coquillages et crustacés crus, transformés ou congelés ainsi qu'à leurs produits dérivés suivants (voir l'Annexe A pour la classification des principaux taxons):
a)    Produits de la pêche, mollusques, tuniciers et échinodermes crus, notamment:
-      poissons entiers ou en filets, avec ou sans peau et/ou tête, vidés;
-      crustacés, entiers ou décortiqués;
-      céphalopodes;
-      mollusques bivalves;
-      gastéropodes;
-      tuniciers et échinodermes.
b)    Produits transformés, notamment:
-      poissons fumés, entiers ou en filets, avec ou sans peau;
-      crustacés entiers ou décortiqués, mollusques, tuniciers et échinodermes cuits, ou partiellement cuits;
-      poissons et produits hétérogènes à base de poissons, cuits ou partiellement cuits.
c)    Poissons, crustacés, mollusques et autres, congelés crus ou cuits, en bloc ou autres, notamment:
-      poissons, filets et morceaux de poisson;
-      crustacés entiers et décortiqués (par exemple, chair de crabe, crevettes), mollusques, tuniciers et échinodermes.
NOTE 2       L'objet des analyses menées sur ces échantillons peut être une analyse d'hygiène ou un contrôle qualité. Cependant, les techniques de prélèvement décrites dans le présent document relèvent davantage de l'analyse d'hygiène (sur tissus musculaires).

Mikrobiologija v prehranski verigi - Priprava preskusnih vzorcev, osnovne suspenzije in decimalnih razredčin za mikrobiološko preiskavo - 3. del: Posebna pravila za pripravo rib in ribjih izdelkov (ISO 6887-3:2017)

Ta dokument določa pravila za pripravo vzorcev rib in ribjih izdelkov ter njihove razredčine za mikrobiološko preiskavo, kadar se za vzorce zahteva drugačna priprava od
metod, opisanih v standardu ISO 6887-1. Standard ISO 6887-1 določa splošna pravila za pripravo osnovnih razredčin za mikrobiološko preiskavo.
Ta dokument zajema posebne postopke za vzorčenje surovih školjk, plaščarjev in iglokožcev na področjih primarne proizvodnje.
OPOMBA 1: vzorčenje surovih školjk, plaščarjev in iglokožcev na področjih primarne proizvodnje je zajeto v tem dokumentu namesto v standardu ISO 13307, ki določa pravila za vzorčenje v kopenski fazi primarne proizvodnje.
Ta dokument ne zajema priprave vzorcev za preskusni metodi za ugotavljanje števila in odkrivanje, pri čemer so podrobnosti glede priprave opredeljene v ustreznih mednarodnih standardih (npr. standarda ISO/TS 15216-1 in ISO/TS 15216-2 za določevanje virusa hepatitisa A in norovirusa v hrani z RT-PCR v realnem času).
Ta dokument je treba uporabljati v povezavi s standardom ISO 6887-1. Uporablja se za naslednje surove, predelane ali zamrznjene ribe in školjke ter njihove izdelke (glej dodatek A za razvrščanje glavnih taksonov):
a) Surovi ribji izdelki, školjke, plaščarji in iglokožci, vključno s/z:
– celimi ribami ali fileji, s kožo in glavami ali brez, brez drobovja;
– raki, celimi ali izluščenimi;
– glavonožci;
– dvolupinskimi školjkami;
– polži;
– plaščarji in iglokožci.
b) Predelani izdelki, vključno s/z:
– prekajenimi ribami, celimi ali pripravljenimi fileji, s kožo ali brez;
– kuhanimi ali delno kuhanimi, celimi ali izluščenimi raki, školjkami, plaščarji in iglokožci;
– kuhanimi ali delno kuhanimi ribami in večkomponentnimi izdelki iz rib.
c) Surove ali kuhane zamrznjene ribe, raki, školjke in ostalo, v blokih ali drugače, vključno s/z:
– ribami, ribjimi fileji in kosi;
– celimi ali izluščenimi raki (npr. koščki rakovice, kozice), školjkami, plaščarji in iglokožci.
OPMBA 2: namen izvedenih preiskav teh vzorcev je lahko testiranje higiene ali nadzor kakovosti. Vendar se tehnike vzorčenja, opisane v tem dokumentu, nanašajo v glavnem na testiranje higiene (na tkivih školjk).

General Information

Status
Published
Publication Date
04-Apr-2017
Current Stage
6060 - Definitive text made available (DAV) - Publishing
Due Date
05-Apr-2017
Completion Date
05-Apr-2017

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SLOVENSKI STANDARD
SIST EN ISO 6887-3:2017
01-junij-2017
1DGRPHãþD
SIST EN ISO 6887-3:2003
0LNURELRORJLMDYSUHKUDQVNLYHULJL3ULSUDYDSUHVNXVQLKY]RUFHYRVQRYQH
VXVSHQ]LMHLQGHFLPDOQLKUD]UHGþLQ]DPLNURELRORãNRSUHLVNDYRGHO3RVHEQD
SUDYLOD]DSULSUDYRULELQULEMLKL]GHONRY ,62

Microbiology of the food chain - Preparation of test samples, initial suspension and

decimal dilutions for microbiological examination - Part 3: Specific rules for the

preparation of fish and fishery products (ISO 6887-3:2017)
Mikrobiologie der Lebensmittelkette - Vorbereitung von Untersuchungsproben und

Herstellung von Erstverdünnungen und von Dezimalverdünnungen für mikrobiologische

Untersuchungen - Teil 3: Spezifische Regeln für die Vorbereitung von Fisch und
Fischerzeugnissen (ISO 6887-3:2017)

Microbiologie de la chaîne alimentaire - Préparation des échantillons, de la suspension

mère et des dilutions décimales en vue de l'examen microbiologique - Partie 3: Règles

spécifiques pour la préparation des produits de la pêche (ISO 6887-3:2017)
Ta slovenski standard je istoveten z: EN ISO 6887-3:2017
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
SIST EN ISO 6887-3:2017 en

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

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SIST EN ISO 6887-3:2017
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SIST EN ISO 6887-3:2017
EN ISO 6887-3
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2017
EUROPÄISCHE NORM
ICS 07.100.30 Supersedes EN ISO 6887-3:2003
English Version
Microbiology of the food chain - Preparation of test
samples, initial suspension and decimal dilutions for
microbiological examination - Part 3: Specific rules for the
preparation of fish and fishery products (ISO 6887-
3:2017)

Microbiologie de la chaîne alimentaire - Préparation Mikrobiologie der Lebensmittelkette - Vorbereitung

des échantillons, de la suspension mère et des dilutions von Untersuchungsproben und Herstellung von

décimales en vue de l'examen microbiologique - Partie Erstverdünnungen und von Dezimalverdünnungen für

3: Règles spécifiques pour la préparation des produits mikrobiologische Untersuchungen - Teil 3: Spezifische

de la pêche (ISO 6887-3:2017) Regeln für die Vorbereitung von Fisch und
Fischerzeugnissen (ISO 6887-3:2017)
This European Standard was approved by CEN on 14 January 2017.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,

Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, 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: Avenue Marnix 17, B-1000 Brussels

© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 6887-3:2017 E

worldwide for CEN national Members.
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SIST EN ISO 6887-3:2017
EN ISO 6887-3:2017 (E)
Contents Page

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

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SIST EN ISO 6887-3:2017
EN ISO 6887-3:2017 (E)
European foreword

This document (EN ISO 6887-3:2017) has been prepared by Technical Committee ISO/TC 34 "Food

products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”

the secretariat of which is held by DIN.

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 October 2017, and conflicting national standards shall

be withdrawn at the latest by October 2017.

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent

rights.
This document supersedes EN ISO 6887-3:2003.

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, Former Yugoslav Republic of Macedonia,

France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,

Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and the United Kingdom.
Endorsement notice

The text of ISO 6887-3:2017 has been approved by CEN as EN ISO 6887-3:2017 without any

modification.
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SIST EN ISO 6887-3:2017
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SIST EN ISO 6887-3:2017
INTERNATIONAL ISO
STANDARD 6887-3
Second edition
2017-03
Microbiology of the food chain —
Preparation of test samples, initial
suspension and decimal dilutions for
microbiological examination —
Part 3:
Specific rules for the preparation of
fish and fishery products
Microbiologie de la chaîne alimentaire — Préparation des
échantillons, de la suspension mère et des dilutions décimales en vue
de l’examen microbiologique —
Partie 3: Règles spécifiques pour la préparation des produits de la pêche
Reference number
ISO 6887-3:2017(E)
ISO 2017
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SIST EN ISO 6887-3:2017
ISO 6887-3:2017(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2017, Published in Switzerland

All rights reserved. Unless otherwise specified, 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
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2017 – All rights reserved
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SIST EN ISO 6887-3:2017
ISO 6887-3:2017(E)
Contents Page

Foreword ..........................................................................................................................................................................................................................................v

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

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

3 Terms and definitions ..................................................................................................................................................................................... 2

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

5 Diluents .......................................................................................................................................................................................................................... 2

6 Apparatus ..................................................................................................................................................................................................................... 2

7 Sampling and sample types ....................................................................................................................................................................... 3

7.1 General procedures ............................................................................................................................................................................. 3

7.2 Specific procedures for sampling bivalve molluscs, echinoderms and tunicates

from primary production ............................................................................................................................................................... 3

7.2.1 General...................................................................................................................................................................................... 3

7.2.2 Sampling and laboratory sample transport ............................................................................................. 3

7.2.3 Sampling method .................. .................................................... ....................................................................................... 3

7.2.4 Size and number of individuals per sample ............................................................................................. 4

7.2.5 Temperature control during transport ......................................................................................................... 4

7.3 Specific procedures for sampling bivalve molluscs, gastropods, echinoderms and

tunicates placed on the market................................................................................................................................................. 4

8 General procedures ........................................................................................................................................................................................... 4

9 Specific procedures............................................................................................................................................................................................ 4

9.1 Raw fishery products, including fish, crustaceans, molluscs, tunicates and

echinoderms (see Annex A) ......................................................................................................................................................... 4

9.1.1 Whole fresh fish (more than 15 cm in length) ....................................................................................... 4

9.1.2 Whole fresh fish (less than 15 cm in length) ........................................................................................... 5

9.1.3 Sliced fish, fillets and steaks ................................................................................................................................... 5

9.1.4 Whole and sliced cephalopods ............................................................................................................................ 5

9.1.5 Whole crustacea such as crabs ............................................................................................................................ 5

9.1.6 Shelled crustacea flesh ............................................................................................................................................... 5

9.1.7 Crustacea such as prawns, crayfish, and lobsters ............................................................................... 6

9.1.8 Live bivalve molluscs .................................................................................................................................................... 6

9.1.9 Echinoderms........................................................................................................................................................................ 7

9.2 Processed products ............................................................................................................................................................................. 7

9.2.1 Whole smoked fish ......................................................................................................................................................... 7

9.2.2 Smoked fish fillets and slices, with or without skin .......................................................................... 8

9.2.3 Whole cooked molluscs in the shell ................................................................................................................ 8

9.2.4 Fish and fish-based multi-component products (e.g. pre-prepared fish

taco, mixed seafood selections, mixed fish ball) ................................................................................... 8

9.2.5 Cooked or precooked shelled bivalves .......................................................................................................... 8

9.2.6 Salted or pickled products (including fish eggs/roe such as caviar) .................................. 8

9.2.7 Dried fish including dried salted fish ............................................................................................................. 9

9.2.8 Fermented products...................................................................................................................................................... 9

9.2.9 Marinated products ....................................................................................................................................................... 9

9.2.10 Breaded products ............................................................................................................................................................ 9

9.3 Frozen fish, crustacea, molluscs, tunicates, and echinoderms ...................................................................... 9

9.3.1 Fish fillets, large fish pieces frozen in blocks, frozen small parts and

single portions ................................................................................................................................................................... 9

9.3.2 Shelled crustacea (such as prawns) frozen in blocks ....................................................................... 9

9.3.3 Whole crustacea (such as prawns) frozen in blocks ......................................................................... 9

9.3.4 Flaked crustacean flesh (such as crab meats) frozen in blocks ............................................... 9

9.3.5 Molluscs (whole cephalopods, bivalve molluscs and gastropods) ....................................10

10 Further dilutions ...............................................................................................................................................................................................10

© ISO 2017 – All rights reserved iii
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SIST EN ISO 6887-3:2017
ISO 6887-3:2017(E)

Annex A (informative) Classification of major taxa ...........................................................................................................................11

Annex B (informative) Recommended number of individual live bivalve molluscs to be

submitted to the laboratory ...................................................................................................................................................................12

Annex C (informative) Additional guidance for small fish, crabs and lobsters .....................................................13

Bibliography .............................................................................................................................................................................................................................16

iv © ISO 2017 – All rights reserved
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SIST EN ISO 6887-3:2017
ISO 6887-3:2017(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 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 . i so .org/ iso/ foreword .html.

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

Microbiology.

This second edition cancels and replaces the first edition (ISO 6887-3:2003), which has been technically

revised.
A list of all parts in the ISO 6887 series can be found on the ISO website.
© ISO 2017 – All rights reserved v
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SIST EN ISO 6887-3:2017
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SIST EN ISO 6887-3:2017
INTERNATIONAL STANDARD ISO 6887-3:2017(E)
Microbiology of the food chain — Preparation of test
samples, initial suspension and decimal dilutions for
microbiological examination —
Part 3:
Specific rules for the preparation of fish and fishery
products

WARNING — The use of this document may involve hazardous materials, operations and

equipment. It is the responsibility of the user of this document to establish appropriate safety

and health practices and to determine the applicability of regulatory limitations before use.

1 Scope

This document specifies rules for the preparation of fish and fishery product samples and their

suspension for microbiological examination when the samples require a different preparation from the

methods described in ISO 6887-1. ISO 6887-1 defines the general rules for the preparation of the initial

suspension and dilutions for microbiological examination.

This document includes special procedures for sampling raw molluscs, tunicates and echinoderms from

primary production areas.

NOTE 1 Sampling of raw molluscs, tunicates and echinoderms from primary production areas is included in this

document, rather than ISO 13307, which specifies rules for sampling from the terrestrial primary production stage.

This document excludes preparation of samples for both enumeration and detection test methods where

preparation details are specified in the relevant International Standards (e.g. ISO/TS 15216-1 and

ISO/TS 15216-2 for determination of hepatitis A virus and norovirus in food using real-time RT-PCR).

This document is intended to be used in conjunction with ISO 6887-1. It is applicable to the following raw,

processed or frozen fish and shellfish and their products (see Annex A for classification of major taxa):

a) Raw fishery products, molluscs, tunicates and echinoderms including:
— whole fish or fillets, with or without skin and heads, and gutted;
— crustaceans, whole or shelled;
— cephalopods;
— bivalve molluscs;
— gastropods;
— tunicates and echinoderms.
b) Processed products including:
— smoked fish, whole or prepared fillets, with or without skin;

— cooked or partially cooked, whole or shelled crustaceans, molluscs, tunicates and echinoderms;

— cooked or partially cooked fish and fish-based multi-component products.
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c) Raw or cooked frozen fish, crustaceans, molluscs and others, in blocks or otherwise, including:

— fish, fish fillets and pieces;

— whole and shelled crustacean (e.g. flaked crab, prawns), molluscs, tunicates and echinoderms.

NOTE 2 The purpose of examinations performed on these samples can be either hygiene testing or quality

control. However, the sampling techniques described in this document relate mainly to hygiene testing (on

muscle tissues).
2 Normative references

The following documents are referred to in 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 6887-1, Microbiology of the food chain — Preparation of test samples, initial suspension and decimal

dilutions for microbiological examination — Part 1: General rules for the preparation of the initial

suspension and decimal dilutions

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

microbiological examinations
3 Terms and definitions

For the purposes of this document, the terms and definitions given in ISO 6887-1 apply.

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

— IEC Electropedia: available at http:// www .electropedia .org/
— ISO Online browsing platform: available at http:// www .iso .org/ obp
4 Principle

The general principles for sample preparation and subsequent steps are described in ISO 6887-1. This

document describes specific measures for fish and fishery products, including raw, processed and

frozen products.
5 Diluents

Diluents for general use and special purposes are described in ISO 6887-1 and there are no additional

specific requirements for fish and fishery products.
6 Apparatus

Usual microbiological laboratory equipment for general use (ISO 7218 and ISO 6887-1) and in particular,

the following:
6.1 Homogenizer.

6.1.1 Rotary homogenizer (blender), as specified in ISO 7218, but if a large test portion is used, the

equipment should include a 1 l bowl.
6.1.2 Peristaltic homogenizer, as specified in ISO 7218.
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6.2 Sterile instruments suitable for dissecting samples and opening shells (e.g. oyster knives,

hammer, pliers, adjustable vice, oyster cracker, sterile scissors, shellfish picker, winkle picker, scalpels

and butcher’s knives).
6.3 Sterile forceps (small and large), spatulas and spoons.
6.4 Small stiff brush, for scrubbing shells.
6.5 Electric drill, equipped with sterile wood bit (14 mm or 16 mm diameter).

6.6 Sterile gauze sheets, suitable for preventing splintering when breaking up shells.

6.7 Food grade plastic bags with waterproof labels, suitable as sampling containers.

6.8 Gloves, strong, suitable to protect operator from injury.
7 Sampling and sample types
7.1 General procedures

Carry out sampling in accordance with the instructions given in this clause for samples from the

primary production stage (7.2) or products placed on the market (7.3). For products not detailed here,

carry out sampling according to the specific standard appropriate to the product concerned or see

ISO/TS 17728. If specific sampling instructions are not available, it is recommended that agreement be

reached on this subject by the parties concerned.

7.2 Specific procedures for sampling bivalve molluscs, echinoderms and tunicates from

primary production
7.2.1 General

The design and implementation of environmental sampling programmes will affect the results obtained

from microbiological examinations. Where the results of this testing are used in microbiological

monitoring programmes, particularly for official controls such as classification and monitoring of

marine production areas, special consideration should be given to formal recording of sampling plans,

[6]
species selection and spatial and temporal aspects of sampling design.
7.2.2 Sampling and laboratory sample transport

A sampling protocol containing details of sampling methods, cleaning, packing and transport

requirements should be agreed by the parties concerned.
7.2.3 Sampling method

The species under examination should be sampled, as far as possible, using the method employed

for commercial harvesting. Equipment used for sampling shall be restricted to that used for this

purpose. To avoid contamination by micro-organisms adhering to marine sediments, disturbance of

surrounding sediments shall be avoided. Once removed from water and having closed, animals shall

be cleaned by rinsing or scrubbing with clean seawater or fresh potable water. Animals shall not be

re-immersed in water.

Individual laboratory samples shall be placed in separate undamaged food grade plastic bags (6.7) or

equivalent, with waterproof labels containing information to ensure sample traceability.

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Where sampling using the commercial harvesting method is not possible, unprocessed animals

harvested for commercial purpose should be taken periodically as checks to show that results for

laboratory samples collected using the alternative sampling method are acceptable.

7.2.4 Size and number of individuals per sample

Laboratory samples should comprise individuals within the normal commercial size range. A pooled

sample comprising a minimum of 10 animals with a minimum amount of flesh and intravalvular

liquid of 50 g should be used (for very small species such as Donax spp. a minimum amount of 25 g is

permitted). Additional animals shall be collected to allow for a proportion of individuals received at the

laboratory being in a moribund state. Recommended numbers of individuals for some species are given

in Annex B.
7.2.5 Temperature control during transport

The temperature of the sample (either the laboratory sample or the surrounding seawater) should be

recorded immediately after collection.

Transport temperature shall be between 0 °C and 10 °C and the equipment used shall be capable of

achieving this temperature range within 4 h of sample packing and maintaining it for at least 24 h. If

cool packs are used, laboratory samples shall not come into direct contact with their surfaces. Samples

shall not be frozen.

Internal air temperature of the temperature controlled unit shall be recorded on receipt at the

laboratory.

For samples where less than 4 h have elapsed between collection from the production area and receipt

at the laboratory, internal air/sample temperature should be less than the temperature recorded at the

time of sampling.

Microbiological examination should be initiated within 24 h of collection of the sample from the

production area. If testing cannot be initiated within 24 h or if sample temperatures between 0 °C and

10 °C cannot be achieved, data should be generated to verify that the use of alternate transport and

storage conditions does not affect the microbiological content of the sample.

NOTE Studies have shown that E. coli will not significantly increase in mussels (Mytilus edulis) or Pacific

[8]
oysters (Crassostrea gigas) at temperatures of 15 °C or less for up to 48 h.

7.3 Specific procedures for sampling bivalve molluscs, gastropods, echinoderms and

tunicates placed on the market
Use the specific sampling procedures given in 7.2.4.
8 General procedures

All preparations and manipulations shall be carried out using aseptic techniques and sterile equipment

(ISO 7218).
9 Specific procedures
9.1 Raw fishery products, including fish, crustaceans, molluscs, tunicates and
echinoderms (see Annex A)
9.1.1 Whole fresh fish (more than 15 cm in length)

The gills and the anus shall be covered with sterile cotton wool, drenched in alcohol at a volume fraction

of 70 %. Disinfect the surface of the dorsal region (using cotton wool with alcohol at a volume fraction

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