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
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AMENDEMENT 2
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ICS: 07.100.20; 07.100.30

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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

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This document is circulated as received from the committee secretariat.
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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
ISO 11133:2014/DAM 2:2017(E)


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ii  © ISO 2017 – All rights reserved

 ISO 11133:2014/DAM 2:2017(E)


Foreword
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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).
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Any trade name used in this document is information given for the convenience of users and does not
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constitute an endorsement.

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For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment,
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as well as information about ISO's adherence to the World Trade Organization (WTO) principles in the
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Technical Barriers to Trade (TBT) see the following URL: www.iso.org/iso/foreword.html.

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The committee responsible for this document is ISO/34, Food products, Subcommittee SC 9, Microbiology.
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© ISO 2017 – All rights reserved  iii

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 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
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of samples from the food chain and water.

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For the microbiological media and reagents under test, the inoculum used is a subculture of an isolated
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colony. Therefore the method of performance testing for these products is qualitative.
-2 ist
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The shortest permissible incubation time in the International Standard should be used for the positive
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control organism(s), whilst the longest permissible incubation time should be used for the negative
0b l o ar
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control organisms.
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The strains chosen in Table K.1 have been selected preferentially from those already cited in
- 3 h. ul
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ISO 11133:2014. If a suitable strain was not available from this source, a strain from the catalogue of
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organisms compiled by the World Data Centre for Microorganisms (WDCM)[20] has been selected.
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In most cases, more than one control strain has been listed in Table K1 for both positive and negative
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reactions. The user may choose any of the strains cited for positive and negative reactions. Refer to the
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relevant standard for the expected results for the target organism.
70 s:
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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

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- WDCMb
ISO Standard Function Control strainsa Characteristic reactions
dium/ reagent numbers
00024 Positive reaction:

Pseudomonas aeruginosa 00025 Yellow to brick red after

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
Escherichia coli Blue colonies with sur-
00179 rounding medium blue/
Acetate agar green
00090
(Sodium acetate 21567 Growth on acetate
agar) Negative reaction:
Shigella.sonnei 00127
) EW No growth or very weak

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growth, no colour change
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Shigella flexneri 00125

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of the medium (remains

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green)
.it R

01 /f0
ds P

-2 ist
00007
Positive reaction:
33 / s
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1 1 ds
Clostridium perfringens 00080
-1 a r

Purplish colour
nd DA

Acid phosphatase Detection of acid phos-

2 0 1 7 g / s d:

14189
so nd

00174
reagent phatase
0b l o ar
20 f/i ta
sta AN

Negative reaction:
4 d t a nd

Clostridium bifermentans 00079

e 2 / c a ta
ST

No purplish colour
b5 a i l s
- 3 h. ul

Positive reaction:
eh

6 c .i t e F

Vibrio
(
iT

00137 turbidity and violet/pur-

Arginine dihydro- Detection of L‑Arginine fluvialis
21872 ple colour
lase saline medium
b0 ds

dihydrolase
00037 Negative reaction:
6- ar

Vibrio parahaemolyticus
1e d
-4 a n

00185 yellow colour

d0 //st

00009
70 s:
tp

Enterococcus faecalis 00087 Positive reaction:

ht

00176 Tan to black colour in the

00177 surrounding medium
Enterococcus faecium
Bile-aesculin-azide Detection of aesculin 00178
7899-2
agar hydrolysis Aerococcus viridans 00061
00012 Negative reaction:
00013 No tan to black colour in
Escherichia coli
00090 the surrounding medium

00179
a Strain free of choice
b Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact
details [20]
c 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
d Non-toxigenic strain of E.coli serotype O157
e Weak coagulase - producing strain of S.aureus

2  © ISO 2017 – All rights reserved

 ISO 11133:2014/DAM 2:2017(E)


Table A.1 (continued)

Confirmation me- WDCMb
ISO Standard Function Control strainsa Characteristic reactions
dium/ reagent numbers
Positive reaction:
Enhanced zone of
00020 β‑haemolysis at the inter-
CAMP medium with Listeria monocytogenes section of the test strain
00021
Staphylococcus au- 11290-1 with each of the cultures of
Detection of CAMP Staphylococcus aureus and
reus WDCM 00034
11290-2 reaction Rhodococcus equi
and Rhodococcus
equi WDCM 00028
Negative reaction:
Listeria ivanovii 00018
No enhanced zone
L.innocua 00017 No zone
00012
11290-1
Carbohydrate utili- 11290-2 00013 Positive reaction: Change
zation broths with Escherichia coli
Detection of carbohy- 00090 of colour to yellow
different carbohy- 21567
drate fermentation
drates and different 000179
indicators 10273
Negative reaction: no
22964 Proteus mirabilis 00123
change in colour
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00032

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Staphylococcus aureus 00034

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33 / s
Campylobacter jejuni
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00005 Positive reaction: Forma-

1 1 ds

tion of bubbles of oxygen

-1 a r

00020
nd DA

2 0 1 7 g / s d:
so nd

9232 Listeria monocytogenes

0b l o ar

00021
20 f/i ta
sta AN

4 d t a nd

10272-1 Listeria innocua 00017

e 2 / c a ta

Catalase reagent Detection of catalase

ST

b5 a i l s

(3 % hydrogen per- 10272-2 after adding hydrogen Listeria ivanovii 00018
- 3 h. ul

oxide solution) peroxide solution

eh

6 c .i t e F
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11290-1 00009
iT

11290-2 E.faecalis 00087

b0 ds

00176 Negative reaction:

6- ar
1e d

00177 No formation of bubbles

-4 a n

Enterococcus faecium of oxygen

d0 //st

00178
70 s:
tp

Lactobacillus delbreuckii
ht

00121
ssp. bulgaricus
Enterobacter aerogenes 00175 Positive reaction:
cream/pink growth with
Enterobacter cloacae 00083
Citrate agar (Chris- surrounding medium red
tensen’s citrate 21567 Growth on citrate
Shigella sonnei 00127
agar)
Negative reaction: No
00125
Shigella flexneri growth
00126
00012
00013 Positive reaction:
Escherichia coli
Glucose agar 11059 00090 Yellow colour
Production of acid
O-F medium with 21528-1 00179
from glucose
overlay 21528-2 Pseudomonas aeruginosa 00025 Negative reaction:
growth, but no yellow
Pseudomonas fluorescens 00115
colour development
a Strain free of choice
b Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact
details [20]
c 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
d Non-toxigenic strain of E.coli serotype O157
e Weak coagulase - producing strain of S.aureus

© ISO 2017 – All rights reserved  3

ISO 11133:2014/DAM 2:2017(E)


Table A.1 (continued)

Confirmation me- WDCMb
ISO Standard Function Control strainsa Characteristic reactions
dium/ reagent numbers
Positive reaction:

Lactobacillus brevis 00099 Agar layer detaches

itself from the underlying
Glucose MRS broth Detection of CO2 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 Escherichia coli 00014d
medium/ reagent
00090
Tryptone/ tryp-
6579 00179 Positive reaction: Forma-
tophan medium
tion of a red ring within
with indole reagent 16654 00037 10 min
(Kovac’s reagent)
21567 Detection of indole for- Vibrio parahaemolyticus 00138
Tryptone/
mation from tryptophan
) EW
21872 00185
tryptophan saline

2- -
d- 2 6
medium with indole 11866-1 Vibrio cholerae 00203
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reagent (Kovacs
11866-2 Vibrio vulnificus

4- 5c
00139
eh E

reagent)
.it R

01 /f0
Enterobacter aerogenes 00175
ds P

Indole reagent
-2 ist Negative reaction:
33 / s
ar RD

(Kovacs reagent) Citrobacter freundii 00006

1 1 ds

Yellow/brown ring within

-1 a r

Salmonella Typhimurium
nd DA

00031
2 0 1 7 g / s d:
so nd

10 min
0b l o ar

Salmonella Enteritidis
20 f/i ta

00030
sta AN

4 d t a nd
e 2 / c a ta

00012
ST

b5 a i l s

Positive reaction:
- 3 h. ul

00013
eh

Escherichia coli
6 c .i t e F
(

Development of a pink col-

00090
iT

our within a few minutes

Indole reagent Detection of indole for- 00179
(Vracko and Sherris mation on membrane
b0 ds

Enterobacter aerogenes
6- ar

reagent) filters 00175

1e d
-4 a n

Citrobacter freundii 00006 Negative reaction: no pink

d0 //st

Salmonella Typhimurium 00031 colour development

70 s:
tp

Salmonella Enteritidis 00030

ht

00024 Positive reaction:

Pseudomonas aeruginosa 00025 presence of fluorescence

under UV radiation
00026 (360 ± 20) nm
King’s B medium 16266 Detection of fluorescein 00012
00013 Negative reaction: no
Escherichia coli
00090 fluorescence

00179
a Strain free of choice
b Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact
details [20]
c 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
d Non-toxigenic strain of E.coli serotype O157
e Weak coagulase - producing strain of S.aureus

4  © ISO 2017 – All rights reserved

 ISO 11133:2014/DAM 2:2017(E)


Table A.1 (continued)

Confirmation me- WDCMb
ISO Standard Function Control strainsa Characteristic reactions
dium/ reagent numbers
00012
Gram-negative:
KOH 00013
Escherichia coli The colony material be-
(Reagent for Gram 00090
comes stringy
typing test) 17995 Gram Typing 00179
(3% potassium hy- Gram-positive:
droxide solution) 00032
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:

) EW Gas production: Durham

tubes more than one quar-

2- -
d- 2 6
Clostridium perfringens 00007
.ai VI
ter full of gas

am c 0
Detection of lactose fer-
LS (Lactose sulfite)

4- 5c
eh E

7937 mentation and sulfite Iron sulfite production: for-

.it R

01 /f0
medium
reduction mation of black precipitate
ds P

-2 ist
33 / s
ar RD

Negative reaction:
1 1 ds

Clostridium sporogenes
-1 a r

00008
nd DA

Gas production but no

2 0 1 7 g / s d:
so nd

blackening
0b l o ar
20 f/i ta
sta AN

4 d t a nd

Salmonella
e 2 / c a ta

00031
ST

b5 a i l s

Typhimuriumc
Positive reaction: Medium
- 3 h. ul
eh

6579
6 c .i t e F

Salmonella Enteritidisc 00030 remains purple after incu-

(
iT

Lysine decarboxy- bation and is turbid

21567 Enterobacter aerogenes 00175
lase medium Detection of L‑Lysine
b0 ds

19250 Vibrio parahaemolyticus 00185

Lysine decarboxy- decarboxylase (LDC)
6- ar

21872 Proteus mirabilis 00023

1e d

lase saline medium

-4 a n

Negative reaction:
Citrobacter freundii 0006
d0 //st

22964
Medium changes from
70 s:

Cronobacter sakazakii 00214

tp

purple to yellow
ht

Cronobacter muytjensis 00213

Positive reaction:
Bacillus cereus 00001
Detection of spores by Green stained spores
Malachite green
21871 microscopic examina-
oxalate solution Negative reaction:
tion
none —
No green-stained spores
a Strain free of choice
b Make reference to the reference strain catalogue available on http://www.wfcc.info for information on culture collection strain numbers and contact
details [20]
c 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
d Non-toxigenic strain of E.coli serotype O157
e Weak coagulase - producing strain of S.aureus

© ISO 2017 – All rights reserved  5