Micro & Molecular Biology

TRYPTONE SOY AGAR

(ISO 9308-1:2000)

Cat : 1138

For the detection and enumeration of E.coli and others Coliform bacteria FORMULA IN GRAMS PER LITER

Casein Triptic Digest .............. 15.00 Sodium Chloride .................... 5.00

Soy Peptone .......................... 5.00 Bacteriological Agar ............... 15.00

Final pH 7.2 0.1 at 25C

Preparation
Suspend 40 grams of the medium in one liter of distilled water. Mix well and heat agitating frequently to boiling. Distribute into appropriate containers and sterilize at 121C for 15 minutes. Allow the medium to cool to 50C and distribute into Petri dishes.

Uses
TRYPTONE SOY AGAR is used for the quick and standard test for the detection and count of E. coli and other Coliforms by the membrane - filter technique as directed by the ISO 9308-1:2000. It is also a general purpose solid medium, particularly useful for the isolation, sensitivity testing and determination of hemolysis with fastidious microorganisms as it does not contain any sugars. Another use of the Tryptone Soy Agar is for the oxidase test according to the traditional method as described in ISO 9308-1:2000. The two different peptones it contains allow the cultivation of a great variety of aerobic and facultative bacteria and also such organisms as Streptococci, Pneumococci and members of the genera Neisseria, Brucella, Corynebacteria, Listeria, Pasteurella, Vibrio and
Erysipelothrix.

Soy peptone is an enzymatic hydrolisate of the soya bean. Casein and Soy peptones provide the essential nutrients for growth: naturally high concentrations of nitrogen, vitamins, minerals, amino acids and natural sugars from the soya bean for carbon and energy. Sodium chloride maintains the osmotic balance and the Bacteriological agar is the solidifying agent. The filtration membrane is placed in the Tryptone Soy Agar and incubated at 36 2C during 4 - 5 hours. The membrane is then transferred to Tryptone Bile Salts Agar (Cat. 1013) and incubated at 44 0.5C during 19 - 20 hours. All indole positive strains which give red colonies when stained with indole reagent are considered as E. coli. For the preparation of blood-plates for hemolysis studies, add 5 - 10% sterile defibrinated blood to sterilized and cooled to 45C medium.

Bibliography
ISO 9308-1:2.000 Regulation water quality-Detection and count of Escherichia coli and coliform bacteria. Anon. 1987 J. Food Microbiol., 5: 291-296.

MICROBIOLOGICAL TEST
The following results were obtained in the performance of the medium from type cultures after incubation at a temperature of 36 2C and observed after 18 - 24 hours.
Microorganisms
Escherichia coli ATCC 25922 Klebsiella pneumoniae ATCC 13833

Growth Good Good

230

- Dehydrated Culture Media -

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