Microbiology of food, animal feed and water - Preparation, production, storage and performance testing of culture media - Amendment 2 (ISO 11133:2014/Amd 2:2020)

Mikrobiologie von Lebensmitteln, Futtermitteln und Wasser - Vorbereitung, Herstellung, Lagerung und Leistungsprüfung von Nährmedien - Änderung 2 (ISO 11133:2014/Amd 2:2020)

Microbiologie des aliments, des aliments pour animaux et de l'eau - Préparation, production, stockage et essais de performance des milieux de culture - Amendement 2 (ISO 11133:2014/Amd 2:2020)

Mikrobiologija živil, krme in vode - Priprava, izdelava, skladiščenje in preskušanje lastnosti gojišč - Dopolnilo A2 (ISO 11133:2014/Amd 2:2020)

General Information

Status
Published
Public Enquiry End Date
04-Mar-2018
Publication Date
18-Jun-2020
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
08-Jun-2020
Due Date
13-Aug-2020
Completion Date
19-Jun-2020

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SLOVENSKI STANDARD
SIST EN ISO 11133:2014/A2:2020
01-julij-2020

Mikrobiologija živil, krme in vode - Priprava, izdelava, skladiščenje in preskušanje

lastnosti gojišč - Dopolnilo A2 (ISO 11133:2014/Amd 2:2020)

Microbiology of food, animal feed and water - Preparation, production, storage and

performance testing of culture media - Amendment 2 (ISO 11133:2014/Amd 2:2020)

Mikrobiologie von Lebensmitteln, Futtermitteln und Wasser - Vorbereitung, Herstellung,

Lagerung und Leistungsprüfung von Nährmedien - Änderung 2 (ISO 11133:2014/Amd
2:2020)
Microbiologie des aliments, des aliments pour animaux et de l'eau - Préparation,

production, stockage et essais de performance des milieux de culture - Amendement 2

(ISO 11133:2014/Amd 2:2020)
Ta slovenski standard je istoveten z: EN ISO 11133:2014/A2:2020
ICS:
07.100.20 Mikrobiologija vode Microbiology of water
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 11133:2014/A2:2020 en

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

---------------------- Page: 1 ----------------------
SIST EN ISO 11133:2014/A2:2020
---------------------- Page: 2 ----------------------
SIST EN ISO 11133:2014/A2:2020
EN ISO 11133:2014/A2
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2020
EUROPÄISCHE NORM
ICS 07.100.30; 07.100.20
English Version
Microbiology of food, animal feed and water - Preparation,
production, storage and performance testing of culture
media - Amendment 2 (ISO 11133:2014/Amd 2:2020)

Microbiologie des aliments, des aliments pour animaux Mikrobiologie von Lebensmitteln, Futtermitteln und

et de l'eau - Préparation, production, stockage et essais Wasser - Vorbereitung, Herstellung, Lagerung und

de performance des milieux de culture - Amendement Leistungsprüfung von Nährmedien - Änderung 2 (ISO

2 (ISO 11133:2014/Amd 2:2020) 11133:2014/Amd 2:2020)

This amendment A2 modifies the European Standard EN ISO 11133:2014; it was approved by CEN on 21 March 2020.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for inclusion of

this amendment into the relevant 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 amendment 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

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11133:2014/A2:2020 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST EN ISO 11133:2014/A2:2020
EN ISO 11133:2014/A2:2020 (E)
Contents Page

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

---------------------- Page: 4 ----------------------
SIST EN ISO 11133:2014/A2:2020
EN ISO 11133:2014/A2:2020 (E)
European foreword

This document (EN ISO 11133:2014/A2:2020) 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 Amendment to the European Standard EN ISO 11133:2014 shall be given the status of a national

standard, either by publication of an identical text or by endorsement, at the latest by November 2020,

and conflicting national standards shall be withdrawn at the latest by November 2020.

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 11133:2014/Amd 2:2020 has been approved by CEN as EN ISO 11133:2014/A2:2020

without any modification.
---------------------- Page: 5 ----------------------
SIST EN ISO 11133:2014/A2:2020
---------------------- Page: 6 ----------------------
SIST EN ISO 11133:2014/A2:2020
INTERNATIONAL ISO
STANDARD 11133
First edition
2014-05-15
AMENDMENT 2
2020-05
Microbiology of food, animal feed and
water — Preparation, production,
storage and performance testing of
culture media
AMENDMENT 2
Microbiologie des aliments, des aliments pour animaux et de l'eau —
Préparation, production, stockage et essais de performance des
milieux de culture
AMENDEMENT 2
Reference number
ISO 11133:2014/Amd.2:2020(E)
ISO 2020
---------------------- Page: 7 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020

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 2020 – All rights reserved
---------------------- Page: 8 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(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 Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4,

Microbiological methods, and in collaboration with the European Committee for Standardization (CEN)

Technical Committee CEN/TC 463, Microbiology of the food chain, in accordance with the Agreement on

technical cooperation between ISO and CEN (Vienna Agreement).

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.
© ISO 2020 – All rights reserved iii
---------------------- Page: 9 ----------------------
SIST EN ISO 11133:2014/A2:2020
---------------------- Page: 10 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)
Microbiology of food, animal feed and water —
Preparation, production, storage and performance testing
of culture media
AMENDMENT 2
End of 5.2 (before the NOTE)
Add the following sentence:

Annex K gives the test microorganisms to be used for confirmation media and reagents in specified

food and water microbiology International Standards.
5.4.1, first sentence
Replace the sentence with the following text:

Suitable microorganisms for routine performance testing are listed in Annexes E, F and K.

End of 6.6.1
Add the following sentence:
Suitable test organisms are described in Annex K.
6.6.2, after the second sentence
Add the following sentence:
Suitable test organisms are described in Annex K.
Annex K
Add the following text as a new annex.
© ISO 2020 – All rights reserved 1
---------------------- Page: 11 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)
Annex K
(normative)
Performance testing of confirmation media and reagents

This annex specifies control strains for the performance testing of confirmation and characterization

media, reagents, dyes, stains and materials described in standards for the microbiological examination

of samples from the food chain and water.

For the microbiological media and reagents under test, the inoculum used is a subculture of an isolated

colony. Therefore, the method of performance testing for these products is qualitative.

The shortest permissible incubation time specified in the relevant International Standard for the

confirmation or characterization test should be used for the positive control organism(s), while the

longest permissible incubation time should be used for the negative control organism(s).

The strains chosen in Table K.1 have been selected preferentially from those already cited in this

document. If a suitable strain was not available from this source, a strain from the catalogue of

[20]

organisms compiled by the World Data Centre for Microorganisms (WDCM) has been selected.

In most cases, more than one control strain has been listed in Table K.1 for both positive and negative

reactions. The user may choose any of the strains cited for positive and negative reactions.

If control strains for performance testing of confirmation or characterization media, reagents, dyes,

stains and materials are already specified in the International Standard, for example, as in ISO 10272-1

and ISO 10272-2 (Campylobacter) and ISO 10273 (Yersinia enterocolitica), they have not been included in

Table K.1. In addition, serological reagents have not been included.

If commercially sourced media or reagents are used, follow the manufacturer’s instructions, including

time, temperature and conditions of performance. If the instructions do not include control strains,

choose a positive and a negative strain from Table K.1. See Clause 6 for requirements.

2 © ISO 2020 – All rights reserved
---------------------- Page: 12 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)

Table K.1 — Control strains for confirmation and characterization media, reagents, dyes,

stains and materials included in documents
from ISO/TC 34/SC 9, ISO/TC 34/SC 5 and ISO/TC 147/SC 4
International WDCM Characteristic
Medium/reagent Function Control strains
Standard numbers reactions
00024
Positive reaction:
Pseudomonas aeruginosa 00025
Yellow to brick red after adding
1 to 2 drops of Nessler’s reagent
00026
Acetamide broth Detection of ammonia
with Nessler’s ISO 16266 production from 00012
reagent acetamide
00013 Negative reaction:
Escherichia coli
00090 No yellow to brick red colour
00179
00012
Positive reaction:
00013
Escherichia coli
Blue colonies with surrounding
00090
Acetate agar medium blue/green
Growth on acetate
00179
ISO 21567
(Sodium acetate
agar
agar)
Shigella sonnei 00127 Negative reaction:
No growth or very weak growth,
Shigella flexneri 00125 no colour change of the medium
(remains green)
00007
Positive reaction:
Clostridium perfringens 00080
Mauve/purple/violet colour
Acid phosphatase Detection of acid
ISO 14189 00174
reagent phosphatase
Negative reaction:
Clostridium bifermentans 00079
No mauve/purple/violet colour
Positive reaction:
Vibrio fluvialis 00137
Turbidity and violet/purple
Arginine
Detection of L-Arginine
colour
dihydrolase saline ISO 21872-1
dihydrolase
medium
00037 Negative reaction:
Vibrio parahaemolyticus
00185 Yellow colour
00009
00087
Enterococcus faecalis
Positive reaction:
00176
Tan to black colour in the
surrounding medium
00177
Enterococcus faecium
00178
Bile aesculin azide Detection of aesculin
ISO 7899-2
agar hydrolysis
Aerococcus viridans 00061
Negative reaction:
00012
No tan to black colour in the
00013
Escherichia coli surrounding medium
00090
00179
Strain free of choice; one of the strains has to be used as a minimum.
b [20]

Refer to the reference strain catalogue available on http:// www .wfcc .info for information on culture collection strain numbers and contact details .

Some national restrictions and directions can require the use of a different serovar. Refer to national requirements relating to the choice of Salmonella

serovars.
Weak coagulase-producing strain of S. aureus.
© ISO 2020 – All rights reserved 3
---------------------- Page: 13 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)
Table K.1 (continued)
International WDCM Characteristic
Medium/reagent Function Control strains
Standard numbers reactions
00012
00013 Positive reaction:
Escherichia coli
00090 Gas produced in Durham tube
ISO 4831
Brilliant green Detection of gas
00179
lactose bile broth production
ISO 4832
00009
Negative reaction:
Enterococcus faecalis 00087
No gas produced in Durham tube
00176
Positive reaction:
00020
Narrow enhanced zone of
Listeria monocytogenes
β-haemolysis at the intersection
00021
of the test strain with
CAMP medium with
Staphylococcus aureus.
Staphylococcus
ISO 11290-1 Wide arrowhead zone of
aureus Detection of CAMP
Listeria ivanovii 00018 β-haemolysis at the intersection
WDCM 00034 and reaction
ISO 11290-2
with Rhodococcus equi
Rhodococcus equi
WDCM 00028
Negative reaction:
Listeria ivanovii 00018
Zone not enhanced with
Staphylococcus aureus
Listeria innocua 00017 No zone
00012
00013
Positive reaction:
Escherichia coli
00090
Change of colour to yellow
ISO 11290-1
00179
Carbohydrate
ISO 11290-2
utilization broths
Detection of
with different
00021
ISO 21567 carbohydrate
carbohydrates and Listeria monocytogenes Rhamnose: yellow
fermentation
00109
different
ISO 10273
indicators
Negative reaction:
ISO 22964
Proteus mirabilis 00023
No change in colour
00021
Listeria monocytogenes Xylose; no change
00109
Strain free of choice; one of the strains has to be used as a minimum.
b [20]

Refer to the reference strain catalogue available on http:// www .wfcc .info for information on culture collection strain numbers and contact details .

Some national restrictions and directions can require the use of a different serovar. Refer to national requirements relating to the choice of Salmonella

serovars.
Weak coagulase-producing strain of S. aureus.
4 © ISO 2020 – All rights reserved
---------------------- Page: 14 ----------------------
SIST EN ISO 11133:2014/A2:2020
ISO 11133:2014/Amd.2:2020(E)
Table K.1 (continued)
International WDCM Characteristic
Medium/reagent Function Control strains
Standard numbers reactions
00032
Staphylococcus aureus
00034
Campylobacter jejuni 00005
Positive reaction:
00020
Listeria monocytogenes
Formation of bubbles of oxygen
00021
ISO 9232
ISO 10272-1 Listeria innocua 00017
Catalase reagent Detection of catalase
(3 % hydrogen ISO 10272-2 after adding hydrogen
peroxide solution) peroxide solution Listeria ivanovii 00018
ISO 11290-1
00009
ISO 11290-2
00087
Enterococcus faecalis
00176
Negative reaction:
00177 No formation of bubbles of oxygen
Enterococcus faecium
00178
Lactobacillus delbrueckii
00121
subsp. bulgaricus
Enterobacter aerogenes 00175 Positive reaction:
Cream/pink growth with
Enterobacter cloacae 00083
surrounding medium red
Citrate agar
Growth on citrate
(Christensen’s ISO 21567 Shigella sonnei 00127
agar
citrate agar)
Negative reaction:
00125
No growth
Shigella flexneri
00126
00012
00013 Positive reaction:
Escherichia coli
00090 Yellow colour
ISO/TS 11059
Glucose agar
00179
Production of acid from
00024
glucose
ISO 21528-1
O-F medium with
00025 Negative reaction:
overlay
ISO 21528-2 Pseudomonas aeruginosa
Growth, but no yellow colour
00026
development
Pseudomonas fluorescens 00115
Pos
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Microbiology of food, animal feed and water - Preparation, production, storage and

performance testing of culture media - Amendment 2 (ISO 11133:2014/DAmd 2:2017)

Mikrobiologie von Lebensmitteln, Futtermitteln und Wasser - Vorbereitung, Herstellung,

Lagerung und Leistungsprüfung von Nährmedien (ISO 11133:2014/DAM 2:2017)
Microbiologie des aliments, des aliments pour animaux et de l'eau - Préparation,

production, stockage et essais de performance des milieux de culture - Amendement 2

(ISO 11133:2014/DAmd 2:2017)
Ta slovenski standard je istoveten z: EN ISO 11133:2014/prA2
ICS:
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 11133:2014/oprA2:2018 en

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

---------------------- Page: 1 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
---------------------- Page: 2 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
DRAFT AMENDMENT
ISO 11133:2014/DAM 2
ISO/TC 34/SC 9 Secretariat: AFNOR
Voting begins on: Voting terminates on:
2017-12-12 2018-03-06
Microbiology of food, animal feed and water —
Preparation, production, storage and performance testing
of culture media
AMENDMENT 2

Microbiologie des aliments, des aliments pour animaux et de l'eau — Préparation, production, stockage et

essais de performance des milieux de culture
AMENDEMENT 2
ICS: 07.100.20; 07.100.30
Member bodies are requested to consult relevant national interests in ISO/TC
147/SC 4 before casting their ballot to the e-Balloting application.
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 11133:2014/DAM 2:2017(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 2017
---------------------- Page: 3 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(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
---------------------- Page: 4 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(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.iso.org/iso/foreword.html.

The committee responsible for this document is ISO/34, Food products, Subcommittee SC 9, Microbiology.

© ISO 2017 – All rights reserved iii
---------------------- Page: 5 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
---------------------- Page: 6 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(E)
Microbiology of food, animal feed and water —
Preparation, production, storage and performance testing
of culture media
AMENDMENT 2
Annex K
(normative)
Performance testing of confirmation media and reagents
K.1 General

This Annex specifies control strains for the performance testing of confirmation and characterization

media, reagents, dyes, stains and materials described in standards for the microbiological examination

of samples from the food chain and water.

For the microbiological media and reagents under test, the inoculum used is a subculture of an isolated

colony. Therefore the method of performance testing for these products is qualitative.

The shortest permissible incubation time in the International Standard should be used for the positive

control organism(s), whilst the longest permissible incubation time should be used for the negative

control organisms.

The strains chosen in Table K.1 have been selected preferentially from those already cited in

ISO 11133:2014. If a suitable strain was not available from this source, a strain from the catalogue of

[20]

organisms compiled by the World Data Centre for Microorganisms (WDCM) has been selected.

In most cases, more than one control strain has been listed in Table K1 for both positive and negative

reactions. The user may choose any of the strains cited for positive and negative reactions. Refer to the

relevant standard for the expected results for the target organism.

If control strains for performance testing of confirmation media and reagents are already specified

in the International Standard, for example as in ISO 10272 (Campylobacter) and ISO 10273 (Yersinia

enterocolitica), they have not been included in Table K.1. In addition, serological reagents have not been

included.

If commercially-sourced media or reagents are used, follow the manufacturer’s instructions, including

time, temperature and conditions of performance. If the instructions do not include control strains,

choose a positive and a negative strain from Table K.1.
© ISO 2017 – All rights reserved 1
---------------------- Page: 7 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(E)

Table A.1 — Control strains for confirmation media and reagents included in ISO standards

from ISO/TC 34/SC 9, ISO/TC34/SC 5 and ISO/TC 147/SC 4
Confirmation me- WDCM
ISO Standard Function Control strains Characteristic reactions
dium/ reagent numbers
Positive reaction:
00024
Yellow to brick red after
Pseudomonas aeruginosa 00025
adding 1-2 drops of Ness-
00026
ler’s reagent
Acetamide broth Detection of ammonia
with Nessler’s 16266 production from aceta-
00012
reagent mide
Negative reaction:
00013
Escherichia coli
No yellow to brick red
00090
colour
00179
00012
Positive reaction:
00013
Blue colonies with sur-
Escherichia coli
00179 rounding medium blue/
Acetate agar
green
00090
21567 Growth on acetate
(Sodium acetate
Negative reaction:
agar)
Shigella.sonnei 00127
No growth or very weak
growth, no colour change
Shigella flexneri 00125
of the medium (remains
green)
00007
Positive reaction:
Clostridium perfringens 00080
Purplish colour
Acid phosphatase Detection of acid phos-
14189
00174
reagent phatase
Negative reaction:
Clostridium bifermentans 00079
No purplish colour
Positive reaction:
Vibrio
00137
turbidity and violet/pur-
Detection of L-Arginine
fluvialis
Arginine dihydro-
ple colour
21872
lase saline medium
dihydrolase
00037 Negative reaction:
Vibrio parahaemolyticus
00185 yellow colour
00009
Enterococcus faecalis 00087
Positive reaction:
00176
Tan to black colour in the
surrounding medium
00177
Enterococcus faecium
00178
Bile-aesculin-azide Detection of aesculin
7899-2
agar hydrolysis
Aerococcus viridans 00061
00012
Negative reaction:
00013
No tan to black colour in
Escherichia coli
the surrounding medium
00090
00179
Strain free of choice

Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact

details [20]

Some national restrictions and directions may require the use of a different serovar. Make reference to national requirements relating to the choice of

Salmonella serovars
Non-toxigenic strain of E.coli serotype O157
Weak coagulase - producing strain of S.aureus
2 © ISO 2017 – All rights reserved
---------------------- Page: 8 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(E)
Table A.1 (continued)
Confirmation me- WDCM
ISO Standard Function Control strains Characteristic reactions
dium/ reagent numbers
Positive reaction:
Enhanced zone of
00020
β-haemolysis at the inter-
Listeria monocytogenes
CAMP medium with section of the test strain
00021
with each of the cultures of
Staphylococcus au-
11290-1
Detection of CAMP
Staphylococcus aureus and
reus WDCM 00034
reaction
11290-2
Rhodococcus equi
and Rhodococcus
equi WDCM 00028
Negative reaction:
Listeria ivanovii 00018
No enhanced zone
L.innocua 00017 No zone
00012
11290-1
00013
Carbohydrate utili-
Positive reaction: Change
11290-2
Escherichia coli
zation broths with
of colour to yellow
Detection of carbohy- 00090
different carbohy- 21567
drate fermentation
drates and different
000179
10273
indicators
Negative reaction: no
22964 Proteus mirabilis 00123
change in colour
00032
Staphylococcus aureus 00034
00090
Campylobacter jejuni 00005
Positive reaction: Forma-
tion of bubbles of oxygen
00020
Listeria monocytogenes
9232
00021
10272-1
Listeria innocua 00017
Catalase reagent Detection of catalase
(3 % hydrogen per- 10272-2 after adding hydrogen
Listeria ivanovii 00018
oxide solution) peroxide solution
11290-1
00009
11290-2
E.faecalis 00087
00176 Negative reaction:
00177 No formation of bubbles
Enterococcus faecium
of oxygen
00178
Lactobacillus delbreuckii
00121
ssp. bulgaricus
Enterobacter aerogenes 00175 Positive reaction:
cream/pink growth with
Enterobacter cloacae 00083
surrounding medium red
Citrate agar (Chris-
tensen’s citrate 21567 Growth on citrate
Shigella sonnei 00127
agar)
Negative reaction: No
00125
growth
Shigella flexneri
00126
00012
00013 Positive reaction:
Escherichia coli
11059
Glucose agar 00090 Yellow colour
Production of acid
21528-1
O-F medium with 00179
from glucose
overlay
21528-2
Pseudomonas aeruginosa 00025 Negative reaction:
growth, but no yellow
Pseudomonas fluorescens 00115
colour development
Strain free of choice

Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact

details [20]

Some national restrictions and directions may require the use of a different serovar. Make reference to national requirements relating to the choice of

Salmonella serovars
Non-toxigenic strain of E.coli serotype O157
Weak coagulase - producing strain of S.aureus
© ISO 2017 – All rights reserved 3
---------------------- Page: 9 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(E)
Table A.1 (continued)
Confirmation me- WDCM
ISO Standard Function Control strains Characteristic reactions
dium/ reagent numbers
Positive reaction:
Agar layer detaches
Lactobacillus brevis 00099
itself from the underlying
Glucose MRS broth Detection of CO pro-
contents
9232
with overlay agar duction
Negative reaction:
Lactobacillus delbruckei
00102
no gas production, agar
ssp. bulgaricus
layer not detached.
00012
00013
Indole reaction d
Escherichia coli 00014
medium/ reagent
00090
Tryptone/ tryp-
6579 Positive reaction: Forma-
00179
tophan medium
tion of a red ring within
with indole reagent
16654
00037
10 min
(Kovac’s reagent)
21567
Vibrio parahaemolyticus 00138
Detection of indole for-
Tryptone/
mation from tryptophan
21872
00185
tryptophan saline
11866-1
Vibrio cholerae 00203
medium with indole
reagent (Kovacs
11866-2 Vibrio vulnificus 00139
reagent)
Enterobacter aerogenes 00175
Indole reagent
Negative reaction:
(Kovacs reagent) Citrobacter freundii 00006
Yellow/brown ring within
Salmonella Typhimurium 00031
10 min
Salmonella Enteritidis 00030
00012
Positive reaction:
00013
Escherichia coli
Development of a pink col-
00090
our within a few minutes
Indole reagent Detection of indole for-
00179
(Vracko and Sherris mation on membrane
Enterobacter aerogenes 00175
reagent) filters
Citrobacter freundii 00006
Negative reaction: no pink
colour development
Salmonella Typhimurium 00031
Salmonella Enteritidis 00030
Positive reaction:
00024
presence of fluorescence
Pseudomonas aeruginosa 00025
under UV radiation
00026
(360 ± 20) nm
King’s B medium 16266 Detection of fluorescein
00012
00013
Negative reaction: no
Escherichia coli
fluorescence
00090
00179
Strain free of choice

Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact

details [20]

Some national restrictions and directions may require the use of a different serovar. Make reference to national requirements relating to the choice of

Salmonella serovars
Non-toxigenic strain of E.coli serotype O157
Weak coagulase - producing strain of S.aureus
4 © ISO 2017 – All rights reserved
---------------------- Page: 10 ----------------------
SIST EN ISO 11133:2014/oprA2:2018
ISO 11133:2014/DAM 2:2017(E)
Table A.1 (continued)
Confirmation me- WDCM
ISO Standard Function Control strains Characteristic reactions
dium/ reagent numbers
00012
Gram-negative:
00013
KOH
Escherichia coli
The colony material be-
00090
(Reagent for Gram
comes stringy
typing test) 17995 Gram Typing
00179
(3% potassium hy-
Gram-positive:
00032
droxide solution)
Staphylococcus aureus
Colony material remains
00034
smooth
Positive reaction:
Clostridium perfringens 00007
Presence of gas, yellow col-
Detection of gas forma-
our and gelatin liquefaction
Lactose-gelatin
7937 tion, acid formation and
medium
Negative reaction:
gelatin liquefaction
Hafnia alvei 00095
No colour change or red,
no gelatin liquefaction
Positive reaction:
Gas production: Durham
tubes more than one quar-
Clostridium perfringens 00007
ter full of gas
Detection of lactose fer-
LS (Lactose sulfite)
7937 mentation and sulfite
Iron sulfite production: for-
medium
reduction mation of black precipitate
Negative reaction:
Clostridium sporogenes 00008
Gas production but no
blackening
Salmonella
00031
Typhimurium
Positive reaction: Medium
6579 c
remains purple after incu-
Salmonella Enteritidis 00030
bation and is turbid
Lysine decarboxy-
21567
Enterobacter aerogenes 00175
lase medium
Detection of L-Lysine
19250 Vibrio parahaemolyticus 00185
decarboxylase (LDC)
Lysine decarboxy-
Proteus mirabilis 00023
21872
lase saline medium
Negative reaction:
Citrobacter freundii 0006
22964
...

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