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Guide for Bio-Rad Products in Food Testing

FoodSafety

Bio-Rad Laboratories • Food Science

Routine Testing

Guide for Bio-Rad Products in Food Testing |

Preparation of dilutions Total flora

NF EN ISO 6887-1 (09/1999)

Preparation of test samples, initial suspension and decimal dilutions for microbiological examination

Part 1: General rules for preparation

of dilutions for microbiological

examination

NF EN ISO 4833 (05/2003)

General guidance for the enumeration of micro-organisms

Colony-count technique at 30°C

dilution 10-1

(and following dilutions)

m g or v ml of sample

+

9 x m g or 9 x v ml of

Tryptone Salt or BufferedPeptone Water

(and following dilutions)

Enumeration

PCA

Initial suspension

and/or

decimal dilutionsOrdering Information

Catalog # Description

PCA (Plate Count Agar)

Ready to use355-4459 100 ml x 6 bottles

355-4457 200 ml x 6 bottles

356-3989 90 mm x 20 dishes

Dehydrated356-4475 500 g

Tryptone Salt/Broth

Ready to use355-5748 9 ml x 40 flip tubes

355-5754 9 ml x 25 tubes

355-5756 90 ml x 6 bottles

355-5791 2.3 l x 5 bags


Buffered Peptone Water/Broth


355-5789 2.3 l x 5 bags

355-5790 5 l x 2 bags


30°C

72 hr

3

PART 1 - ROUTINE TESTING

Preparation of dilutions 3

Total flora 3

Escherichia coli - ISO standards 4

Escherichia coli - Alternative method:

RAPID’E.coli 2 Agar 5

Coliforms - ISO standards 6

Coliforms - Alternative method:

RAPID’E.coli 2 Agar 7

Enterobacteriaceae 8-9

Mesophilic lactic acid bacteria 10

Yeasts and Molds 11

PART 2 - PATHOGEN TESTING

Salmonella - ISO standards 12

Salmonella - Alternative method:

RAPID'Salmonella Agar 14

Salmonella - Alternative method :

iQ-Check™ Salmonella II real-time PCR kit 15

Listeria monocytogenes - ISO standards 16-17

Listeria monocytogenes - Alternative method:

RAPID’L.mono Agar 18

Listeria monocytogenes - Alternative method :

iQ-Check™ Listeria monocytogenes II real-time

PCR kit 19

Listeria spp. - Alternative method :

RAPID’Listeria spp. Agar 20

Listeria spp. - Alternative method :

iQ-Check™ Listeria spp. real-time PCR kit 21

Escherichia coli O157:H7 - Alternative method:

RAPID’E.coli O157:H7 Agar 22

Escherichia coli O157:H7 - Alternative method:

iQ-Check™ E.coli O157:H7 real-time PCR kit 23

Staphylococcus aureus - ISO standards 24

Staphylococcus aureus - Alternative method:

RAPID’Staph Agar 25

Sulfite-reducing bacteria 26

Clostridium perfringens 26

Bacillus cereus 27

Shigella 27

Contents

Guide for Bio-Rad Products in Food Testing

FoodSafety

THIS BOOKLET IS ONLY AN INDICATION PROVIDED BY BIO-RAD FOR THE USE OF BIO-RAD PRODUCTS.IT IS NOT A CONTRACTUAL DOCUMENT.

ISO 16649-1 (04/2001)

Horizontal method for the enumeration of β-glucuronidase-positive E. coli

Part 1: Colony-count technique at

44°C using membranes and

5-bromo-4-chloro-3-indolyl-

β-D-Glucuronide

TBX

Direct enumeration

Sample/

initial suspension

MMGA with membrane

ISO 16649-2 (04/2001)

Horizontal method for the enumerationof β-glucuronidase-positive E. coli

Part 2: Colony-count technique at

44°C using 5-bromo-4-chloro-

3-indolyl-β-D- Glucuronide

Escherichia coli - ISO standards

ISO 7251 (07/2005)

General guidance for detection and enumeration

of presumptive Escherichia coli

Most probable number technique

ISO/TS 16649-3 (09/2005)

Horizontal method for the enumerationof β-glucuronidase-positive E. coli

Part 3: Most probable number

technique using 5-bromo-4-chloro-3-

indolyl-β-D-Glucuronide

Enumeration of Escherichia coliAFNOR validation according to ISO 16140

N° Bio-Rad 07/1-07/93 BRD 07/7-12/04

AOAC-RI approved N° 050601

NordVal approved

All food stuffs

Dilutions

Resuscitation

Sample/

initial suspension

RAPID’E.coli 2

Dilutions

TBX

Direct enumeration

Sample/

initial suspension

Dilutions

Tryptose LaurylSulfate

EC Medium

Peptone Water

Indole test reading and calculation

Sample/

initial suspension

Single

concentration

Double

concentration

Routine Testing

5| Guide for Bio-Rad Products in Food Testing4

Ordering Information


TBX/Agar



Peptone Water/Broth

Ready to use355-4180 9 ml x 25 tubes


Tryptose Lauryl Sulfate/Broth


EC Medium/Broth


RAPID’E.coli 2/Agar



44°C

18-24 hr

Room t° 15 min

and 37°C

4 hr ± 1 hr

44°C

18-24 hr37°C

24-48 hr

± 2 hr

37°C

24-48 hr

± 2 hr

44°C

24-48 hr

± 2 hr

44°C/37°C

18-24 hr

Escherichia coli Alternative method: RAPID’E.coli 2/Agar

Direct Enumeration

of typical colonies*

* Enumeration of typical colonies for E. coli at 44°C; for E. coli and coliforms at 37°C

Sample/initial suspension

* For detection method only

(or)*

44°C

48 hr

± 2 hr

Each tube showing the presence of blue

colonies on TBXshould be considered positive

Calculate the most probable number

from the number of positive tubes

at each dilution

44°C

20-24 hr

37°C

24 hr ± 2 hr

Add 10 mlin 3 tubes of double

strength MMG

And subsequent dilutionAdd 1 ml in 3 tubes

of single strength MMG

Subculture each tube presenting

yellow coloration on a dish of TBX

Guide for Bio-Rad Products in Food Testing |

* Please refer to ISO 7218: 2007

(2 X)*

Routine Testing

76

ISO 4831 (08/2006)

Microbiology of food and animal feeding stuffs Horizontal method for the detection

and enumeration of coliforms MPN technique

Sample

Tryptose Lauryl Sulfate

ISO 4832 (02/2006)

Microbiology of food and animalfeeding stuffs

Horizontal methodfor the enumeration of coliforms

Colony-count technique

Coliforms - ISO standards

ColiformsAlternative method: RAPID’E.coli 2/Agar

Enumeration of coliformsAFNOR validation according to ISO 16140

N° Bio-Rad 07/8-12/04

AOAC-RI approved N° 050601

NordVal approved

All food stuffs

Dilutions

Direct enumeration

of typical colonies*

Sample/

initial suspension

RAPID’E.coli 2

Dilutions

VRBL

Enumeration

of characteristic colonies

Sample/

initial suspension

Dilutions

* Enumeration of typical colonies for E. coli at 44°C; for E. coli and coliforms at 37°C



BGBLB (Brillant Green Bile

Lactose Broth)



VRBL/Agar

(Violet Red Bile Lactose)



Tryptose Lauryl Sulfate/Broth


RAPID’E.coli 2/Agar



30°C or 37°C

24-48 hr ± 2 hr

30°C or 37°C

24 hr ± 2 hr

30-37°C

24 hr (+24 hr)

30-37°C

24 hr ± 2 hr

37°C

18-24 hr

Single

concentration

Double

concentration

Reading (gas +)

Coliforms

• Diameter ≥ 0.5 mm

• Purple with or without

a red zone of precipitated bile

BGBLB

(or)*


Guide for Bio-Rad Products in Food Testing | | Guide for Bio-Rad Products in Food Testing


(2 X)*

ISO 21528-1 (08/2004)

Microbiology of food and animal feeding stuffs Horizontal methods for the detection

and enumeration of Enterobacteriaceae

Part 1: Detection and enumeration by MPN technique

with pre-enrichment

ISO 21528-2 (08/2004)

Microbiology of food and animal feeding stuffsHorizontal methods for the detection

and enumeration of Enterobacteriaceae

Part 2: Colony-count technique

VRBG

Oxydase Dextrose BCP

yellow colour (+)

EE

Biochemical confirmation

Confirmation

Nutrient (2.8%)

Sample

Routine Testing

Reading

Non selective Pre-enrichment

Dilutions

Selective enrichment

Oxydase (-) Dextrose BCP

yellow colour (+)

Buffered Peptone Water

Biochemical confirmation

Confirmation

Nutrient (2.8%)


VRBG

Enterobacteriaceae

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr

37°C

18 hr ± 2 hr

98



EE/Broth


VRBG/Agar

(Violet Red Bile glucose)



Nutrient (2.8%)/Agar



Oxydase Test

Cartridge of 30 disks355-3832



355-5789 2.3 l x 5 bags

355-5790 5 l x 2 bags



(2 X)*


Routine Testing

ISO 15214 (08/1998)

Microbiology of food and animal feeding stuffsHorizontal method for enumeration of Mesophilic Lactic Acid Bacteria


ISO 7954 (08/1988)

General guidance for enumerationof Yeasts and Molds



Dilutions


Mesophilic Lactic Acid Bacteria Yeasts and Molds

30°C

72 hr25°C

3, 4 and 5 days

Enumeration Reading

1110



MRS/Agar

(Man, Rogosa and Sharp)


MRS YGC Ordering Information


YGC/Agar

(Yeast Glucose Chloramphenicol)

Ready to use355-5489 100 ml x 6 vials



(2 X)*


(2 X)*


Pathogen Testing

1312





355-5789 2.3 l x 5 bags

355-5790 5 l x 2 bags


RVS (Rappaport Vassiliadis

Soy)/Broth



MKTTn/Broth



Novobiocin supplement356-4610 1 vial

TSI (Triple Sugar Iron)/Agar




XLD (Xylose Lysine

Deoxycholate)/Agar

Ready to use354-1751 90 mm x 20 dishes


Selenite-Cystine/Broth

Complete medium355-5744 20 ml x 25 tubes

Base356-4074 500 g

Sodium biselenite356-4075 100 g

MSRV/Semi-solid Agar



XLT 4

(Xylose Lysine Tergitol 4)/Agar


Brillant Green

(Kristensen)/Agar


Hektoen/Agar


356-3894 90 mm x 20 dishes


RAPID’Salmonella/Agar



Oxydase Test

Cartridge of 30 disks355-3832

Antiserum Salmonella

Omnivalent OMNI-0 (A60)

356-0781 3 ml dropper bottle

EN ISO 6579/A1 (02/2006)

Microbiology of food and animal feeding stuffsHorizontal method for the detection of Salmonella spp.

Amendment 1 Appendix D : detection of Salmonella spp. in animal faeces and in samplesfrom primary production stage

Salmonella - ISO standards

37°C ± 1°C

24 hr ± 3 hr

37°C ± 1°C

24 hr ± 3 hr

37°C ± 1°C

24 hr ± 3 hr

41.5°C ± 1°C

24 hr ± 3 hr

37°C ± 1°C

18 hr ± 2 hr

RappaportVassiliadis Soya


MKTTn

Appendix D Protocol

Biochemical and serological confirmation

TSI Agar and other test

Plating-out

Confirmation

Nutrient 2.1%

Sample

Non selectivepre-enrichment

Selectiveenrichment

Confirmation

Selectiveisolation

XLD RAPID'Salmonellaor XLT 4 or Kristensenor Hektoen

MSRV

41.5°C ± 1.5°C

24 hr ± 3 hr

(+24 hr for negative)


Pathogen Testing

1514



RAPID’Salmonella/Agar





355-5789 2.3 l x 5 bags

355-5790 5 l x 2 bags


RVS (Rappaport Vassiliadis

Soy)/Broth



SalmonellaAlternative methods : RAPID’Salmonella/Agar iQ-Check™ Salmonella II real-time PCR kit

37°C

18 hr

± 2 hr

37°C

18 hr

± 2 hr

37°C

21 hr

± 1 hr

41.5°C

6 to 26 hr

37°C

24 hr

± 2 hr

BufferedPeptone Water

RappaportVassiliadis Soya

RAPID’Salmonella

Sample

Plating-out

Reading & confirmation

Standard tests or

Oxydase/OMNI-O/ONPG


Sample

Non selectivepre-enrichment

Selectiveenrichment

Selectiveisolation

Detection of Salmonella spp.AFNOR validation according to ISO 16140

N° Bio-Rad 07/11-12/05

AOAC-RI approved N° 050701*

All food stuffs and feed

Detection of Salmonella spp.AFNOR validation according to ISO 16140

N° Bio-Rad 07/06 – 07/04

AOAC-RI approval pending*

All food stuffs, feed, and environment

Bacterial Lysis &

Standard DNA

Extraction

Bacterial Lysis &

Easy DNA

Extraction

Real-time PCR amplification and detection

with iQ-Check Salmonella II

Result analysis and interpretation

with iQ-Check Analysis software

or


Purchase of iQ-Check kits includes an immunity from suit under patents specified in the product insert to use only the amount purchasedsolely in food testing, environmental testing, industrial microbiology and also for the purchaser’s own internal research. No other patent rights(such as 5’ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licensesmay be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Oxydase Test

355-3832 Cartridge of 30 disks

ONPG Test

355-3822 Set of 50 disks

Antiserum Salmonella

Omnivalent OMNI-0 (A60)

356-0781 3 ml dropper bottle

iQ-CheckTM Salmonella II Kit

357-8123 96 tests

For more information on Bio-Rad’sreal-time PCR kits and instruments,please contact your local Bio-Radsales representative.

*For AOAC-RI protocol, please contactyour local Bio-Rad sales representative

EN ISO 11290-1/A1 (02/2005)

Horizontal method for the detectionand enumeration of Listeria monocytogenes

Part 1: Detection method

EN ISO 11290-2/A1 (02/2005)

Horizontal method for the detectionand enumeration of Listeria monocytogenes

Part 2: Enumeration method

Enumeration and confirmation tests

Reading andconfirmation

Sample

Fraser 1/2 without LiCl

and selective supplement

or Buffered Peptone Water

Agar Listeria accordingto Ottaviani and Agosti

Dilution

Listeria spp.

- TSYEA agar

- Catalase +

- Gram

- Mobility

Listeriamonocytogenes- TSYEB

- Hemolysis

- Carbohydrates

- Camp test

Isolation

Primary enrichment

Isolation Secondary enrichment

Sample

Fraser 1/2

Fraser 1(0.1 ml culture/10 ml)

Listeria spp.

- TSYEA agar

- Catalase +

- Gram

- Mobility

Listeriamonocytogenes- TSYEB

- Hemolysis

- Carbohydrates

- Camp test

Agar Listeria accordingto Ottaviani and Agosti

Other solidselective medium

Pathogen Testing

1716

Listeria monocytogenes - ISO standards

30°C

24 hr ± 2 hr

20°C ± 2°C

1 hr ± 5 min.

37°C

24 hr ± 3 hr

(+ 24 hr ± 3 hr)35°C or 37°C

48 hr ± 2 hr




Ready to use355-4179 225 ml x 6 bottles355-5789 2.3 l x 5 bags355-5790 5 l x 2 bagsDehydrated356-4684 500 g

Fraser 1/2/Broth

Complete medium355-5797 225 ml x 6 bottles355-5788 2.3 l x 5 bagsBase356-4604 500 gSupplement356-4616 10 vials

Fraser 1/Broth

Complete medium355-4569 10 ml x 25 tubesBase356-4604 500 gSupplement356-4615 10 vials

AL/Agar (Agar Listeria according

to Ottaviani and Agosti)

Ready to use356-3695 90 mm x 20 dishesBase356-4038 500 gSupplement 1356-4039 10 vialsSupplement 2356-4040 10 vials

Palcam/Agar

Complete medium356-3674 90 mm x 20 dishesBase356-4754 500 gSupplement356-4752 10 vials

Oxford/Agar

Complete medium356-3664 90 mm x 20 dishes

37°C ± 1°C

24 hr ± 3 hr

(+24 hr ± 3 hr)

30-35-37°C(depending

of the media)

24-48hr ± 2 hr

Reading andconfirmation

37°C ± 1°C

24 hr ± 3 hr

(+24 hr ± 3 hr)

30-35-37°C(depending

of the media)

24-48hr ± 2 hr

Agar Listeria

accordingto Ottaviani and Agosti

Other solidselectivemedium


Detection of Listeria monocytogenesAFNOR validation according to ISO 16140

n° Bio-Rad 07/4-09/98

AOAC-RI approved N°030406*

NordVal approved

All food stuffs and environment

Pathogen Testing

Listeria monocytogenesAlternative methods: RAPID’L.mono/Agar

30°C

24 hr

± 2 hr

37°C

24 hr

± 2 hr

1918

Isolation on

RAPID’L.mono

Reading and

confirmation tests

Fraser 1/2 broth

Sample

37°C

24 hr

± 2 hr

Isolation on

RAPID’L.mono

Direct

enumeration


Sample

Standard Tests

- TSYEB

- Hemolysis

- Carbohydrates

- Camp test

Spot on

Agar Listeriaaccording to

Ottaviani and

Agosti

or

or

or



RAPID’L.mono/Agar

Ready to use356-3694 90 mm x 20 dishes355-5294 190 ml + suppl.



Fraser 1/2/Broth

Complete medium355-5797 225 ml x 6 bottles355-5788 2.3 l x 5 bagsBase356-4604 500 gSupplement356-4616 10 vials

AL/Agar (Agar Listeria according

to Ottaviani and Agosti)

Ready to use356-3695 90 mm x 20 dishesBase356-4038 500 gSupplement 1356-4039 10 vialsSupplement 2356-4040 10 vials

LSB (Listeria Special Broth)


iQ-CheckTM

Listeria monocytogenes II Kit

357-8124 96 tests

For more information on Bio-Rad’s real-timePCR kits and instruments, please contactyour local Bio-Rad sales representative.

*For AOAC-RI protocol, please contactyour local Bio-Rad sales representative

Enumeration of Listeria monocytogenesAFNOR validation according to ISO 16140

N° Bio-Rad 07/05-09/01


20°C

1 hr

Resuscitation

For an AFNOR validated enumeration method, in the event of a positive result using RAPID’L.mono for enumeration, confirmation is not necessary when the presence of Listeria monocytogeneshas been confirmed during detection.

iQ-Check™ Listeria monocytogenes II real-time PCR kit

30°C

25 hr

± 1 hr

30°C

25 hr

± 1 hr

30°C

23 hr

± 1 hr

Listeria Special Broth (LSB) Fraser 1/2 broth

Sample

Detection of Listeria monocytogenesAFNOR validation according to ISO 16140

N° Bio-Rad 07/10 – 04/05

AOAC-RI approval pending*


Bacterial Lysis & Standard DNA Extraction Bacterial Lysis &

Easy DNA Extraction

Real-time PCR amplification and detection with

iQ-Check Listeria monocytogenes II


with iQ-Check Analysis softwareConfirmation tests with RAPID’L.mono


L. mono L. ivanonii L. innocua L. welshimeri

Purchase of iQ-Check kits includes an immunity from suit under patents specified in the product insert to use onlythe amount purchased solely in food testing, environmental testing, industrial microbiology and also for the purcha-ser’s own internal research. No other patent rights (such as 5’ Nuclease Process patent rights) are conveyedexpressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contac-ting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Pathogen Testing

2120

Reading and confirmation tests

Isolation on RAPID’Listeria spp.

Sample

Fraser 1/2 broth

Standard Tests

- Gram

- Catalase

Spot on

RAPID'L.mono+ Gram test

+ Catalase

Spot on Palcam

30°C

24 hr ± 2 hr

37°C

24 hr ± 2 hr



RAPID’Listeria spp./Agar

Ready to use356-3950 20 x 90 mm dishesDehydrated Base356-4744 500 gSupplement 1356-4745 1 box, 10 vials

(1 vial qsp 500 ml)Supplement 2356-4746 1 box, 10 vials

(1 vial qsp 500 ml)

Fraser 1/2/Broth

Ready to use355-5797 225 ml x 6 bottles355-5788 2.3 l x 5 bagsBase356-4604 500 gSupplement356-4616 10 vials

Palcam/Agar

Ready to use356-3674 90 mm x 20 dishesBase356-4754 500 gSupplement356-4752 10 vials

LSB (Listeria Special Broth)


iQ-CheckTM Listeria spp. Kit

357-8113 96 tests

For more information on Bio-Rad’s real-timePCR kits and instruments, please contactyour local Bio-Rad sales representative.

*For AOAC-RI protocol, please contact yourlocal Bio-Rad sales representative

iQ-Check™ Listeria spp. real-time PCR kit

Listeria spp.Alternative method s:RAPID’Listeria spp./Agar

30°C

23 hr

± 1 hr

30°C

25 hr

± 1 hr

Listeria Special Broth (LSB)

Sample

Detection of Listeria spp.AFNOR validation according to ISO 16140

N° Bio-Rad 07/13 – 05/07




N° Bio-Rad 07/12 - 12/06



Bacterial Lysis &

Standard DNA

Extraction

Bacterial Lysis &

Easy DNA

Extraction


with iQ-Check Listeria spp.


with iQ-Check Analysis softwareConfirmation tests with RAPID’Listeria spp. or RAPID’L.mono


or

| Guide for Bio-Rad Products in Food Testing Guide for Bio-Rad Products in Food Testing |

or or


N° Bio-Rad 07/4-09/98


30°C

24 hr

± 2 hr

37°C

24-48 hr

Isolation on

RAPID’L.mono

Reading and

confirmation tests

Fraser 1/2 broth

Sample

Standard Tests

- TSYEA

- Gram

- Catalase

Pathogen Testing

2322



RAPID’E.coli O157:H7/Agar


mTSB/Broth

Ready to use (containing novobiocin)355-5426 225 ml x 6 bottles

Dehydrated (base)356-4426 500 g

Novobiocin




iQ-CheckTM E.coli O157:H7 Kit

357-8114 96 tests

For more information on Bio-Rad’sreal-time PCR kits and instruments,please contact your local Bio-Radsales representative.

*For AOAC-RI protocol, pleasecontact your local Bio-Rad salesrepresentative

Escherichia coli O157:H7Alternative methods: RAPID’E.coli O157:H7/Agar

41.5°C

16-24 hr

mTSB + Novobiocin

Immuno-magnetic-separation

Sample

37°C

24 hr ± 2 hr

Reading and confirmation tests

Standard tests or Latex

Isolation on RAPID’E.coli O157:H7

Detection of Escherichia coli O157:H7AFNOR validation according to ISO 16140

N° Bio-Rad 07/14-09/07



RAPID’E.coli O157:H7 can also be used as a second mediumin the ISO 16654 standard


iQ-Check™ E.coli O157:H7 real-time PCR kit

41.5°C

8-24 hr


Sample

Detection of Escherichia coli O157:H7AOAC-RI approval pending*

All food stuffs

Bacterial Lysis &

Easy DNA Extraction


with iQ-Check E.coli O157:H7


with iQ-Check Analysis software


NF EN ISO 6888-2/A1 (12/2003)

Horizontal method for the enumeration ofcoagulase-positive staphylococci

(Staphylococcus aureus and species)

Technique using Rabbit plasma

Fibrinogen agar medium

NF EN ISO 6888-3 (06/2003)

Horizontal method for the enumerationof coagulase-positive staphylococci

(Staphylococcus aureus and species)

Detection and MPN technique

for low numbers

35°C or 37°C

18-24 hr

and 48 hr

37°C

24 hr ± 2 hr

(+24 hr ± 2 hr)

37°C

24-48 hr

37°C

4-6 hr

or 24 hr

37°C

24 h

37°C

24-48 hr

Direct enumeration of typical

coagulase-positive staphylococci

Sample/initial suspensionand following dilution

Rabbit Plasma

Confirmation of

typical colonies

on BHI

Reading

Direct

enumeration

of typical

coagulase-

positive colonies


Giolitti and CantoniBroth

Simple concentration

Baird- Baird-ParkerParker for RPF

+ RPF sup.

37°C

24 hr ± 2 hr

RAPID’Staph


Enumeration and Confirmation

Staphylococcus aureusAlternative method:RAPID’Staph/Agar

2524

NF EN ISO 6888-1/A1 (01/2004)

Horizontal method for the enumeration of coagulase-positive staphylococci

(Staphylococcus aureus and other species)

Technique including confirmation

of colonies

Staphylococcus aureus - ISO standards

35°C or 37°C

24 hr ± 2 hr

(+ 24 hr ± 2 hr)

35°C or 37°C

24 hr ± 2 hr

35°C or 37°C

4-6 hr or 24 hr

Confirmation

Brain-Heart Infusion

Rabbit plasma

Coagulation reading

Sample/initial suspensionand following dilution

Enumeration

Typical staphylococciGrey to black colonies with a clear halo around

and a white opaque zone



Baird-Parker/Agar

Complete medium356-3991 90 mm x 20 dishes

Dehydrated base356-4814 500 g

Baird-Parker for RPF/Agar

Complete medium357-8618 90 ml x 6 bottles

+ 6 vials of suppl.356-3996 90 mm x 20 dishesDehydrated base356-4814 500 g

Additives

Egg yolk with potassium tellurite355-4201 5 ml x 1 vial

355-4205 25 ml x 1 bottle

Sulphamethazine solution (0.2%)356-2682 2.5 ml x 1 vial

Pastorex™ Staph Plus Latex

355-6356 50 tests

355-6353 5 x 50 tests

RPF Supplement

356-4618 10 vials

Brain-Heart Infusion

(BHI)/Broth



Rabbit Plasma

Freeze-dried355-6352 Pack for 20 reactions

Special broth355-3544 9 ml x 25 tubes

RAPID’Staph/Agar


Dehydrated base356-4704 500 g

Baird-Parker Baird-Parkerfor RPF

+ RPF sup.

or

Gioletti and CantoniBroth

Double concentration(or)*


Enumeration of Coagulase-positiveStaphylococcus

AFNOR validation according to ISO 16140N° Bio-Rad 07/9-02/05

AOAC-RI approved N° 080602*All food stuffs, feed and environment

Pathogen Testing


(2 X)*


(2 X)*


Pastorex TM

Staph Plus

Spot on

Baird-Parker

+ RPF

or

*For AOAC-RI protocol, pleasecontact your local Bio-Rad salesrepresentative

ISO 15213 (05/2003)

Microbiology of food and animal feeding stuffsHorizontal method for

the enumeration of sulfite-reducingbacteria growing

under anaerobic conditions

Reading

Food sample/initial suspension

Dilution

Confirmation

NF EN ISO 7937 (02/2005)

Horizontal method for the enumerationof Clostridium perfringens

Colony-count technique

Sulfite-reducing bacteria Clostridium perfringens Bacillus cereus

NF EN ISO 7932 (07/2005)

General guidance for the enumerationof Bacillus cereus


Thermophilousbacteria

suspicion50°C 24-48 hr

Anaerobes condition

Confirmation test

on 5 well isolated colonies


LactoseSulfite

Pathogen Testing

2726



TSC (Tryptose Sulfite

Cycloserin)/Agar

(without D-Cycloserin)



Lactose Sulfite/Broth

Base356-4914 100 g

Thioglycolate +

Resazurin/Broth



Columbia + Sheep Blood/Agar



Sheep Blood Supplement355-6652 50 ml vial

37°C

24-48 hr

anaerobes

condition

Heat

treatment

20 mn 75°C

37°C

20 hr ± 2 hr

30°C

24 hr ± 2 hr

37°C

18-24 hr

37°C

18-24 hr

46°C

24 hr

30°C

18-24 hr

(+24 hr)

Enumeration

of typical black colonies

Confirmation

of 5 isolated colonies with

Thioglycolate + Resazurin


MobilityNitrate

LactoseGelatine

Clostridiumperfringens(yellow/gas+)

Clostridiumperfringens(red coloration/

gas+)

Clostridiumperfringens(black precipitate/

gas+)

Anaerobescondition Columbia + Sheep blood

Reading and interpretation



Mossel (Mannitol egg

Yolk Polymixin)/Agar


XLD (Xylose Lysine

Deoxycholate)/Agar



Mac Conkey (with Crystal

Violet)/Agar



Hektoen/Agar


356-3894 90 mm x 20 dishes




Novobiocin


TSC

TSC(add D-Cycloserine)

Mossel (MYP)

Shigella

NF EN ISO 21567 (03/2005)

Horizontal methodfor the detection of Shigella spp.

37°C

20-24 hr

(+48 hr)

37°C

20-24 hr

41°C

16-20 hr

Plating-out

Shigella Broth + NovobiocinPH 7 ± 0.2

Spot on typical colonies on

Nutrient (2.1%)


Biochemical and serologicalconfirmation

MacConkey

XLD Hektoen


(2 X)*


(2 X)*


(2 X)*


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