Tiêu chuẩn iso 06887 3 2017

© ISO 2017 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 an[.]

Microbiolog y ofthe food chain —

Part 3:

Specific rules for the pr eparation of

Micro iolo ie de la ch îne alimentaire — Pré aration de s

éch ntillo s,de la sus pension mè e et de s d utions décimale s en v e

de l’e xamen micro iolo ique —

Partie  3: Rè gle s sp cifiq e s p ur la pré aratio  de s produit de  la pêche

COPYRIGHT PROTECTED DOCUMENT

© ISO 2017, P blshed in Sw itz rlan

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written permis ion Permis ion c n be req esed from either ISO at the ad r es below or ISO’s member bod y in the c u try of

F reword v

1 Sc ope 1

2 Nor mati ve r eferenc es 2

3 Terms and definitions 2

4 Principle 2

5 Diuents 2

6 A pparatus 2

7 S mpl n and sample types 3

7.1 General pr oc d res

3 7.2 Spe ific pr oc d r es for samplng biv lve moluscs, e hinode ms and tu icates fr om primary prod ction 3

7.2.1 General 3

7.2.2 Samplng an la oratory sample tr ansp rt 3

7.2.3 Samplng method 3

7.2.4 Siz an n mbe of individ als pe sample 4

7.2.5 Temperatur e contr ol d ring transport 4

7.3 Spe ific pr oc d r es for samplng biv lve moluscs, gas r op ds, e hinode ms an tunicates plac d on the mar ket .4

8 General proc edur es 4

9 Specific pr oc edures 4

9.1 Raw fishe y pr od cts, inclu ing fish, c ustac ans, moluscs, tunicates an e hinode ms ( e A nnex A) 4

9.1.1 Whole fr esh fish (mor e than 1 cm in leng th) 4

9.1.2 Whole fr esh fish (les than 1 cm in leng th) 5

9.1.3 Slc d fish, fi et an s eaks 5

9.1.4 Whole an slc d c phalo ods 5

9.1.5 Whole c us ac a such as crabs 5

9.1.6 Sheled c ustac a flesh 5

9.1.7 Crus ac a such as pra w ns, cra yfish, an lo s e s 6

9.1.8 Live biv lve moluscs 6

9.1.9 Echinode ms 7

9.2 Pr oc s ed prod ct 7

9.2.1 Whole smoked fish 7

9.2.2 Smoked fish fi ets an slc s, with or without skin 8

9.2.3 Whole co ked moluscs in the shel 8

9.2.4 Fish an fish-based multi-comp nent pr oducts (e.g pr e-pr ep r ed fish taco, mixed seafo d sele tions, mixed fish b l) 8

9.2.5 C oked or preco ked sheled biv lves 8

9.2.6 Salted or pickled prod ct (inclu ing fish eg g s/roesuch asca viar) 8

9.2.7 Dried fish inclu ing dried salted fish 9

9.2.8 Fermented pr od ct 9

9.2.9 Marinated pr od ct 9

9.2.10 Breaded pr od ct 9

9.3 Fr oz n fish, c us ac a, moluscs, tu icates, an e hinode ms 9

9.3.1 Fish fi et , lar g e fish pie es froz n in blocks, froz n smal part an single p rtions 9

9.3.2 Sheled c ustac a ( uch as prawns) fr oz n in blocks 9

9.3.3 Whole c us ac a ( uch as prawns) froz n in blocks 9

9.3.4 Flaked c us ac an flesh ( uch as crab meat )fr oz n in blocks 9

9.3.5 Moluscs (whole c phalo ods, biv lve moluscs an gas r op ds) 1

A nne x A (informative) Clas ificatio of major taxa 11

A nne x B (informative)Rec ommended numbe of indi vidual li ve bi val ve molus s to be

submit ed to the laboratory 12

A nne x C (informative) Additional g uidance for smal fish, crabs and lobster s 13

Biblog raph y 16

ISO (he Int ernational Org nization for Stan ardization) is a worldwide fede ation of national s an ards

b dies (ISO membe b dies) The work of pr p ring Int ernational Standards is normaly car ied out

through ISO t ech ical committ ees Each membe b dy int er st ed in a subje t for w hich a t ech ical

committ ee has be n es a l shed has the right t o be r pr sent ed on that committ ee Int ernational

org nizations, g ove nmental an non-g ove nmental, in laison with ISO, also take part in the work

ISO cola orat es closely with the Int ernational Ele trot ech ical C mmis ion (IEC) on al matt ers of

ele trot ech ical s an ardization

The proc d r s used t o develo this document an those int en ed for it furthe maint enanc ar

desc ibed in the ISO/IEC Dir ctives, Part 1 In p rticular, the dife ent a pro al c it eria ne ded for the

dife ent ty es of ISO document should be not ed This document was draft ed in ac ordanc with the

edit orial rules of the ISO/IEC Dir ctives, Part 2 ( e www iso org dir ctives)

A tt ention is drawn t o the p s ibi ity that ome of the element of this document ma be the subje t of

p t ent right ISO shal not be held r sponsible for identifying any or al such pat ent right Detais of

any p t ent right identif ied d ring the develo ment of the document wi be in the Introd ction an / r

on the ISO ls of p t ent de larations r c ived ( e www iso org p t ent )

Any trade name used in this document is information given for the convenienc of use s an does not

cons itut e an en orsement

For an ex lanation on the meaning of ISO spe ific terms an ex r s ions r lated to conformity as es ment,

as wel as information a out ISO’s adhe enc to the Wor ld Trade Org nization (WTO) principles in the

Te h ical Bar ie s to Trade (TBT) se the folowing URL: www iso org / iso/ for word html

This document was pr p r d b Te hnical Committ ee ISO/TC 34, Fo d produc ts , Subcommitt ee SC 9,

Microbiolog y ofthe food chain — Pr eparation oftest

Part 3:

Specific rules for the pr eparation offish and fishery

WARNING — The use of this document may inv lve hazardous materials, o eratio s and

equipment It is the respo sibiity of the use of this document to establ sh ap ro riate safety

and he lth practices an to determine the ap l cabi ity of re ulatory l mitatio s before use

This document spe ifies rules for the pr p ration of fish an f ishe y prod ct samples an their

suspension for mic o iolo ical ex mination w hen the samplesr q ir a dife ent pr p ration from the

methods desc ibed in ISO 6 87-1 ISO 68 7-1 defines the g ene al rules for the pr p ration of the initial

suspension an diutionsfor mic o iolo ical ex mination

This document inclu es spe ial proc d r s for samplng raw moluscs, tunicat es an e hinode ms from

primary prod ction ar as

NOT 1 S mpling of raw mol us s, tu icat es an echinoderms from primary prod ction are s is inclu ed in this

document, rather than ISO 1 3 7, w hich specifies rules for ampling from the t er estrial primary prod ction stag e

Thisdocument ex clu es pr p ration of samples for b th en me ation an det ection t es methods w he e

pr p ration detais ar spe ified in the r lev ant Int ernational Stan ards (e.g ISO/TS 1 2 6-1 an

ISO/TS 1 2 6- 2 for det ermination of hep titis A virus an noro irus in fo d using r al-time RT-PCR)

This document is int en ed t o be used in conjunction with ISO 6 8 -1 It is a plca le to the folowing raw,

proc s ed or froz n fish an shelfish an their prod ct ( e An ex A for clas ification of major ta a):

a) Raw fishe y prod ct , moluscs, tunicat es an e hinode ms inclu ing:

— w hole f ish or fil et , with or without skin an heads, an gutt ed;

— c us ac ans, w hole or sheled;

— c phalo ods;

— biv lve moluscs;

— g s ro ods;

— tunicat es an e hinode ms

b) Proc s ed prod ct inclu ing:

— smoked f ish, w hole or pr p r d f ilet , with or without kin;

— co ked or p rtialy co ked, w hole or sheled c us ac ans, moluscs, tu icat es an e hinode ms;

— co ked or p rtialy co ked fish an f ish-b sed multi-comp nent prod ct

c) Raw or co ked froz n fish, c us ac ans, moluscsan othe s, in blocks or othe wise, inclu ing:

— f ish, fish fil et an pie es;

— w hole an sheled c us ac an (e.g flaked c a , prawns), moluscs, tu icat es an e hinode ms

NOTE 2 The p rpose of e aminations performed on these samples can b either hy giene t esting or q ality

control However, the sampling t ech iq es des rib d in this document relate mainly to hy giene testing (on

mus le tis ues)

2 Normati ve referenc es

The fol owing document ar r fe r d t o in t ext in such a wa that some or al of their cont ent

cons itut es r q ir ment of this document F or dat ed r fe enc s, only the edition cit ed a ples F or

un at ed r fe enc s, the lat es edition of the r fe enc d document ( inclu ing any amendment ) a ples

ISO 68 7-1, Mic ro iolo y o the fod chain — Prep ration o tes t s amples , initial s us pens io a d dec imal

diutio s fr mic ro iolo ic al ex min tio — Part 1: G ene al rules fr the pr p ratio o the initial

s us pens io a d dec imal diutio s

ISO 7 1 , Mic ro iolo y o f od a d a imal fe din s tufs — G ene al r q ir ments a d g ida ce fr

mic ro iolo ic al ex min tio s

3 Terms and definitions

F or the purposes of this document, the t erms an definitions given in ISO 6 87-1 a ply

ISO an IEC maintain t erminolo ical data ases for use in s an ardization at the folowing ad r s es:

— IECEle tro edia: a aia le at ht p:/ www ele tro edia org

— ISO Onlnebrow sing plat orm:a ai a le at ht p:/ www iso org o p

4 Principle

The g ene al principles for sample pr p ration an subseq ent st eps ar desc ibed in ISO 6 87-1 This

document desc ibes spe if ic measur s for fish an fishe y prod ct , inclu ing raw, proc s ed an

froz n prod ct

Diuent for g ene al use an spe ial purp ses ar desc ibed in ISO 6 8 -1 an the e ar no ad itional

spe ific r q ir ment for f ish an f ishe y prod ct

6 A pparatus

Usual mic o iolo ical la orat ory eq ipment for g ene al use (ISO 7 1 an ISO 68 7-1) an in p rticular,

the folowing:

6.1 Homog enizer

6.1.1 Rotary homog enizer (blende ), as spe ified in ISO 7 1 , but if a lar g e tes p rtion is used, the

eq ipment hould inclu e a 1 l b wl

6.1.2 P er istaltic homog enizer, as spe ified in ISO 7 1

6.2 Ster ile instr uments suitable for dis e ting samples and o ening shels (e.g o s e k ives,

hamme , ple s, adjustable vic , o s e cracke , s e ie scis ors, shelfish picke , w in le picke , scalpels

and butche ’s knives)

6.3 Ster ile for c eps ( mal and lar g e), spatulas an spo ns

6.4 Smal stif brush, for sc ub ing shels

6.5 Electr ic dril, eq ip ed with s e ie wo d bit (14 mm or 1 mm diamete )

6.6 Ster ile gauze she ts, suitable for preventing spl nte ing when br eaking up shels

6.7 F od g rade plastic bag s with water proof labels, suita le as samplng containe s

6.8 Glo es, s r ong , suita le to prote t o er ator fr om injury

7 Sampl ng and sample types

7.1 General pr oc edur es

Car y out sampl ng in ac ordanc with the ins ructions given in this clause for samples from the

primary prod ction s ag e (7.2) or prod ct plac d on the market (7.3) F or prod ct not detai ed he e,

car y out sampl ng ac ording t o the spe if ic s an ard a pro riat e t o the prod ct conc rned or se

ISO/TS 1 7 8 If spe if ic samplng ins ructions ar not a ai a le, it is r commen ed that a r ement be

r ached on thissubje t b the p rtiesconc rned

7.2 Specific proc edures for sampl ng bi val ve mol us s, echinoderms and tunicates from

primary pr oduction

7.2.1 General

The design an implementation of environmental sampl ng pro rammes wi l afe t he r sult o tained

from mic o iolo ical ex minations Whe e the r sult of this t es ing ar used in mic o iolo ical

monit oring pro rammes, p rticularly for official controls such as clas ification an monit oring of

marine prod ction ar as, spe ial conside ation should be given t o formal r cording of sampl ng plans,

spe ies sele tion an sp tial an t emporal aspe t of samplng design

[6]

7.2.2 S mplng and laboratory sample transport

A samplng prot ocol containing detais of samplng methods, cleaning, p cking an transp rt

r q ir ment should be a r ed b the p rties conc rned

7.2.3 S mplng meth d

The spe ies u de ex mination should be sampled, as far as p s ible, using the method employed

for comme cial harves ing Eq ipment used for samplng shal be r s ricted to that used for this

purpose To avoid contamination b mic o-org nisms ad e ing to marine sediment , dis urbanc of

sur oun ing sediment shal be avoided Onc r moved from wate and ha ing closed, animals shal

be cleaned b rinsing or sc ub ing with clean seawate or fr sh pota le wate Animals shal not be

r -imme sed in wate

In ivid al la orat ory samples shal be plac d in separat e u damag ed fo d grade plas ic b gs (6.7) or

eq iv lent, with wat erpro f la elscontaining information t o ensur sample trac a i ty

Whe e sampl ng using the comme cial harves ing method is not p s ible, u proc s ed animals

harvest ed for comme cial purp se should be taken pe iodicaly as che ks t o show that r sult for

la orat ory samples cole t ed using the alt ernative samplng method ar ac epta le

7.2.4 Size and numbe of indi viduals per sample

L b rat ory samples should comprise in ivid als within the normal comme cial siz rang e A p oled

sample comprising a minimum of 1 animals with a minimum amount of flesh and intra alvular

lq id of 5 g should be used (for ve y smal spe ies such as Do a sp a minimum amount of 2 g is

pe mitt ed) Ad itional animals shal becole t ed t o alow for a pro ortion of in ivid als r c ived at he

la orat ory being in a moribu d s at e R ecommended n mbe s of individ als for some spe ies ar given

in An ex B

7.2.5 Temperature c ontrol dur in transpor t

The t empe atur of the sample (eithe the la orat ory sample or the sur oun ing seawat er) should be

r corded immediat ely aft er cole tion

Transp rt t empe atur shal be betwe n 0 °C an 1 °C an the eq ipment used shal be ca a le of

achieving this t empe atur rang e within 4 h of sample p cking an maintaining it for at leas 24 h If

co l packs ar used, la orat ory samples shal not come int o dir ct contact with their surfac s Samples

shal not be froz n

Int ernal air t empe atur of the t empe atur controled u it shal be r corded on r c ipt at the

la orat ory

F or samples w he e les than 4 h ha e ela sed between col e tion from the prod ction ar a an r c ipt

at the la orat ory, int ernal air/sample t empe atur should be les than the t empe atur r corded at the

time of samplng

Mic o iolo ical ex mination should be initiat ed within 24 h of cole tion of the sample from the

prod ction ar a If t es ing cannot be initiat ed within 24 h or if sample t empe atur s between 0 °C an

1 °C can ot be achieved, data should be g ene at ed t o ve ify that the use of alt ernat e transp rt an

st orag e con itions does not afe t the mic o iolo ical cont ent of the sample

NOTE Stu ies have shown that E c ol wil not signif icantly inc e se in mus els ( My tius eduli s) or Pacif ic

oyster (Cr s sos tr a gigas) at t emperatures of 1 °C or les for u t o 48 h

[8]

7.3 Specific pr oc edur es for sampl ng bi val ve mol us s, gastropods, echinoder ms and

tunicates plac ed on the mar ket

Use the spe ific samplng proc d r s given in 7.2.4

8 General pr oc edur es

Al pr p rations an manipulations shal be car ied out using aseptic t ech iq es an st eri e eq ipment

(ISO 7 1 )

9 Specific pr ocedures

9.1 Raw fishery pr oducts, including fish, c ustac eans, mol us s, tunicates and

echinoderms (see A nne x A)

9.1.1 Whole fresh fish (more than 15 cm in leng th)

The gi ls an the an s shal be co e ed with st erie cott on wo l, dr nched in alcohol at a v lume fraction

of 7 % Disinfe t the surfac of the dorsal r gion (using cott on wo l with alcohol at a v lume fraction

of 7 % )and r mo e an discard a se tion of the skin using st erie for eps (6.3) an scalpel (6.2) Take a

cube-sha ed sample of dorsal muscle, dic it an br ak up in an a pro riat e di uent

If the fish is evisc rat ed, the gi ls shal be co e ed with st eri e cott on wo l, dr nched in alcohol at a

v lume fraction of 7 %, an a cube-sha ed sample of dorsal muscle shal be r mo ed from inside the

9.1.2 Whole fresh fish (les than 15 cm in leng th)

Using st erie scis ors (6.2) an for eps (6.3), r mo e a p rtion of f ish jus ant erior t o the tai inse tion

b making two cut t o prod c transve se se tions, the firs cut t o r mo e the tai and tai inse tion

an the se on t o r move a st eak

Use sufficient mat erial from the la orat ory sample t o give a r pr sentative t es portion as spe ified in

the t es method

A dd diuent t o make a 1 in 1 suspension an blen in a rotary or pe is altic homog eniz r (6.1) as

ne es ary

NOTE A dditional guidance for smal f ish u t o 1 cm in length is given in An e C

9.1.3 Slc ed fish, filets and ste ks

No spe ific r q ir ment ; tr at in ac ordanc with ISO 68 7-1

9.1.4 Whole and slc ed c ep alop ds

Disinfe t the surfac of the skin and suckers (using cott on wo l with alcohol at a v lume fraction

of 7 % ) R emo e the skin an suckers with st erie for eps (6.3) an a scalpel (6.3) an discard Take

cube-sha ed samples of dorsal muscles an pie es from the t entacles Use sufficient mat erial from the

la orat ory sample t o give a r pr sentative t es p rtion as spe if ied in the t es method

The flesh from c phalo ods is r latively f irm; grin up the t es p rtion in diuent using a rotary

homog eniz r (6.1.1) or cut it int o fine pie es A dd furthe diuent t o make a 1 in 1 suspension an blen

in a rotary or pe is altic homog eniz r (6.1) asne es ary

9.1.5 Whole c ustac ea such as crabs

Disinfe t the surfac (using cott on wool with alcohol at a v lume fraction of 7 % ) an with st erie

hamme (6.2), ple s (6.2) or for eps (6.3) r mo e or br ak the cara ac ( e C.2) an claw s t o extract

the ma imum amount of flesh for t es ing F or larg e claw s, an oy st er c acke (6.2) can be used t o br ak

o en the shel befor extracting the flesh

Use sufficient mat erial from the la orat ory sample t o give a r pr sentative t es portion as spe ified in

the t es method

A dd diuent t o make a 1 in 1 suspension an blen in a rotary or pe is altic homog eniz r (6.1) as

ne es ary

9.1.6 Sheled c ustac ea flesh

Take the amount of flesh r q ir d in the t es method, make the initial 1 in 1 suspension in a diuent

an blen in a rotary or pe is altic homog eniz r (6.1) as ne es ary

Use suff icient mat erial from the la orat ory sample t o give a r pr sentative t es portion as spe ified in

the t es method

9.1.7 Cr ustac ea such as praw ns, crayfish, and lo ster s

9.1.7.1 Species where tais o l y are c onsumed

Disinfe t he surfac (using cott on wool with alcohol at a volume fraction of 7 % ) Br ak the c us ac an

at the ju ction between the c phalothora an a domen ( e Figur C.3) Using st erie for eps (6.3)pul

the edible p rtion of flesh from thec phalothora an but en of the a domen

Use sufficient mat erial from the edible portion of the la orat ory sample t o give a r pr sentative t es

p rtion as spe ified in the t es method

A dd the ne es ary q antity of diuent t o give a 1 in 1 suspension

Blend in a rotary or pe is altic homog eniz r (6.1)asne es ary

9.1.7.2 Species c onsumed whole

Take the entir animal for ex mination Use suff icient mat erial from the la orat ory sample t o give a

r pr sentative t es p rtion as spe if ied in the t es method

A dd the ne es ary q antity of diuent t o give a 1 in 1 suspension

Blend in a rotary or pe is altic homog eniz r (6.1)asne es ary

9.1.8 Li ve bi val ve molus s

9.1.8.1 General

On ar iv l at the la orat ory, the int ernal air t empe atur of the transp rt containe shal be r corded

F or samples w he e mor than 4 h ha e ela sed betwe n cole tion an r c ipt, the int ernal air

t empe atur should be betwe n 0 °C an 1 °C If the int ernal air t empe atur of the transp rt

containe is gr at er than 1 °C, the sample t empe atur should be measur d; this should not ex ce d

1 °C F or samples w he e les than 4 h ha e ela sed between cole tion an r c ipt, int ernal air/sample

t empe atur should be les than the t empe atur r corded at the time of samplng

L b rat ory samples shal best or d at 3 °C± 2 °C

The animals shal be al ve Discard in ivid als with o en or damag ed shel s A r pr sentative t es

sample shal contain at leas 1 in ivid als

[7]

and shal be at leas 5 g (2 g for smal animals, e.g Do a

sp )asdetaied in 7.2.4 a o e Tes ing of biv lves inclu es b th the flesh an intra alvular wat er; o en

sufficient shel fish t o yield the amou t of flesh an intrav alvular fluid spe ified in the t es method

Mic o iolo ical ex mination should be initiat ed within 24 h of col e tion of the sample If t es ing can ot

be initiat ed within 24 h or if sample t empe atur s of 0 °C and 1 °C can ot be achieved, data should

be g ene at ed t o ve ify that the use of alt ernat e transp rt an st orag e con itions does not afe t the

mic o iolo ical cont ent of the sample

NOTE Stu ies have shown that E c ol wil not signif icantly inc e se in mus els ( My tius eduli s) or Pacif ic

oyster (Cr s sos tr a gigas) at t emperatures of 1 °C or les for u t o 48 h

[8]

9.1.8.2 Metho s r eq iring a 1 in 10 initial suspensio

Wash an brush (6.4) each shel u de run ing wat er of p ta le q alty, espe ialy aroun the hing e or

o ening

Drain the cleaned biv lves an put them on a clean surfac