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PLATE COUNT AGAR (PCA)

INTENDED USE
Plate Count Agar containing glucose and yeast extract is used in alimentary bacteriology to enumerate
aerobic bacteria in milk, meats, meat-based products, other food products, as well as for the analysis of
pharmaceuticals, cosmetics and their associated raw materials.
HISTORY
Plate Count Agar is prepared with the same ingredients as those originally used by Buchbinder et al.
These authors compared several batches of yeast extract and showed that the results obtained
(without milk added to the medium) were satisfactory for the enumeration of bacteria that were
contaminating samples of raw and pasteurized milk. The transparency of the medium and the relative
size of colonies formed led to uncomplicated enumeration.
PRINCIPLES
The nutrients supplied by Tryptone, vitamins from yeast extract, and glucose used as energy source
favor the growth of most bacteria.
PREPARATION
-

Suspend 20.5 g of dehydrated medium (BK144) in 1 liter of distilled or deionized water.


Slowly bring to boiling, with constant agitation until complete dissolution.
Dispense in tubes or flasks.
Sterilize in an autoclave at 121C for 15 minutes.

NOTE :
Incomplete agar melting during preparation will invariably lead to significant inconsistency in the gel
strength of the solidified agar, after sterilization and cooling.

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INSTRUCTIONS FOR USE


- With the ready-to-use media BM015 or BM033 (or if the media has been prepared in advance from
the dehydrated base), melt the medium for the minimum amount of time necessary to achieve total
liquefaction.
- Cool and maintain at 44-47C.
- Transfer 1 mL of the product to analyze and its serial tenfold dilutions to sterile Petri dishes.
- Pour 10 to 15 mL of medium.
- Homogenize by swirling.
- Let solidify on a cold surface.
- If necessary, overlay with 4 mL of sterile agar (Bacteriological Agar type A at 15 g/L).
- Let solidify again.

- Incubate :

at 30C for 72 hours for the detection of mesophilic bacteria.


at 55C for thermophilic bacteria.
at 6.5C for 10 days for psychrophilic bacteria.

NOTE 1 :
In dairy bacteriology, it is recommended to add 1 g of powdered skimmed milk per liter of reconstituted
medium.

RESULTS
Only plates containing less than 300 colonies but greater than 10 are used for enumeration. When
grown on PCA containing milk, caseinolytic bacteria form a clear halo around each colony (proteolysis
of casein in the milk).

TYPICAL COMPOSITION
(can be adjusted to obtain optimal performance)
For 1 liter of medium :
- Tryptone ...........................................................................................5.0 g
- Yeast extract ....................................................................................2.5 g
- Glucose ............................................................................................1.0 g
- Bacteriological agar .......................................................................12.0 g
pH of the ready-to-use medium at 25C : 7.0 0.2.

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QUALITY CONTROL
- Dehydrated medium : cream powder, free-flowing and homogeneous.
- Prepared medium : light amber agar.
- Typical culture response after 72 hours of incubation at 30C :
Growth
(Productivity ratio : PR)

Microorganisms
Escherichia coli
Staphylococcus aureus
Bacillus subtilis

ATCC 25922
ATCC 6538P
ATCC 6633

PR 70%
PR 70%
PR 70%

STORAGE / SHELF LIFE


Dehydrated medium : 2-30C.
- The expiration date is indicated on the label.
Prepared medium (benchmark value*) :
- Media in tubes or vials : 6 months at 2-8C.
- Media in plates : 1 month at 2-8C
Ready-to-melt media in vials :
- Store between 2-8C, shielded from light.
- The expiration date is indicated on the label.

PACKAGING

Code

Ready-to-melt media in vials :


- 10 x 100 mL
- 10 x 200 mL

BM01508
BM03308

Dehydrated medium :
- 500 g bottle
- 5 kg drum

BK144HA
BK144GC

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BIBLIOGRAPHY
Buchbinder, Baris, Alff, Reynolds, Dillon, Pessin, Pincus and Strauss. 1951. Public Health Reports, 66: 327.
Mthode officielle pour le dnombrement des germes arobies msophiles. Ministre de lAgriculture. Commission XXX.
Cosmtologie.
NF T 72-151. Novembre 1988. Antiseptiques et dsinfectants utiliss l'tat liquide, miscibles l'eau. Dtermination de
l'activit bactricide en prsence de substances interfrentes de rfrence. Mthode par filtration sur membranes.
NF ISO 7698 (V 03-763). Aot 1991. Crales, lgumineuses et produits drivs. Dnombrement des bactries, levures et
moisissures.
T 90-425. Fvrier 1992. Essais des eaux. Examens bactriologiques des rcipients et systmes de bouchage destins aux
eaux conditionnes.
NF V 08-051. Fvrier 1999. Microbiologie des aliments. Dnombrement des microorganismes par comptage des colonies
obtenues 30 C. Mthode de routine.
NF ISO 17410 (V 08-033). Novembre 2001. Microbiologie des aliments. Mthode horizontale pour le dnombrement des
micro-organismes psychrotrophes.
NF EN ISO 4833 (V 08-011). Mai 2003. Microbiologie des aliments. Mthode horizontale pour le dnombrement des microorganismes. Technique de comptage des colonies 30 C.
XP CEN ISO/TS 11133-2 (V 08-104-2). Janvier 2004. Microbiologie des aliments. Guide pour la prparation et la production
des milieux de culture. Partie 2 : Guide gnral pour les essais de performance des milieux de culture.
ISO 8553 / IDF 131. Mai 2004. Lait. Dnombrement des micro-organismes. Mthode de lanse sur botes de Petri 30 C.
ISO 8552 / IDF 132. Mai 2004. Lait. Estimation des micro-organismes psychrotrophes. Technique par comptage des colonies
21 C (mthode rapide).

NF EN 14569 (V 03-011). Fvrier 2005. Produits alimentaires. Dpistages microbiologiques des aliments ioniss en utilisant la
technique LAL/GNB.
ISO 14461-1 / IDF 169-1. Mai 2005. Lait et produits laitiers. Contrle de qualit en laboratoires microbiologiques. Partie 1:
Evaluation de la performance des analystes effectuant les comptages de colonies.
ISO 8784-1. Juillet 2005. Pte, papier et carton. Analyse microbienne. Partie 1: Dnombrement total des bactries, levures et
moisissures bas sur la dsintgration.
ISO 6730 / FIL 101. Septembre 2005. Lait. Dnombrement des units formant colonie de micro-organismes psychrotrophes.
Technique par comptage des colonies 6,5 C.

*Benchmark value refers to the expected shelf life when prepared under standard laboratory conditions following manufacturers instructions. It is provided as a
guide only and no warranty, implied or otherwise, is associated with this information.

The information provided on the package take precedence over the formulations or instructions described in this document.
The information and specifications contained in this technical data sheet date from 2010-01-11.
They are susceptible to modification at any time, without warning.
Code document : BK144/A/2003-01 : 8.

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Biokar Diagnostics Rue des Quarante Mines ZAC de Ther Allonne B.P. 10245 F60002 Beauvais Cedex France
Tl : + 33 (0)3 44 14 33 33 Fax : + 33 (0)3 44 14 33 34 www.biokar-diagnostics.com