R2A AGAR (EUROPEAN PHARMACOPOEIA)

CAT N: 1071
FORMULA IN g/l
Proteose Peptone Starch Glucose Yeast Extract Casein Hydrolysate 0.50 0.50 0.50 0.50 0.50 Dipotassium Phosphate Sodium Pyruvate Magnesium Sulfate Anhydrous Bacteriological Agar 0.30 0.30 0.024 15.00

For the total aerobe count in treated waters

Final pH 7.2 0.2 at 25C

PREPARATION
Suspend 18.12 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 121C for 15 minutes. Cool to 50C, mix well and dispense into plates. The prepared medium should be store at 8-15C. The color is amber. The dehydrated medium should be homogeneous, free-flowing and beige in color. If there are any physical changes, discard the medium.

USES
R2A AGAR was developed by Reasoner and Geldreich for bacteriological plate counts of treated potable water, being able to recover the stressed chlorine-treated bacteria. Nutritionally rich mediums suppress these slow growing bacteria, whereas a low nutrient medium, such as R2A Agar, in combination with a lower incubation temperature and longer incubation time, stimulates the growth of stressed and chlorine-tolerant bacteria. Proteose peptone and Casein Hydrolysate provide nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract is the source of vitamins, particularly of the B-group. Dextrose is a source of fermentable carbohydrate as an energy source; Starch absorbs toxic metabolic byproducts and thereby aids the recovery of injured organisms. Sodium pyruvate increases the recovery of stressed cells. Magnesium sulfate provides divalent cations and sulfate. Dipotassium phosphate is used to balance the pH and provide phosphate. Bacteriological agar is solidifying base. Inoculate plates with tap water samples using the streak plate technique and/or membrane filter method. Incubate at 35C 2C for 24-72 hours. R2A Agar is recommended in Standard Methods for the Examination of Water and Wastewater for pour plate, spread plate and membrane filter methods for heterotrophic counts. The European Pharmacopoeia recommends in paragraph Water for Injections (Aqua ad iniectabilia) this medium for growth promotion. Inoculate plates of R2A Agar separately (with no more than 100 CFU) of Pseudomonas aeruginosa ATCC 9027 and Bacillus subtilis ATCC 6633 and incubate at 30-35C 3 days.

MICROBIOLOGICAL TEST
The following results were obtained in the performance of the medium from type cultures after incubation at a temperature of 35 2C and observed after 24-72 hours.

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Microorganisms

Growth Good Good Good Good Good Good Good Good

Escherichia coli ATCC 25922 Escherichia coli ATCC 11775 Escherichia coli ATCC 8739 Staphylococcus aureus ATCC 25923 Staphylococcus aureus ATCC 6538 Staphylococcus epidermidis ATCC 12228 *Pseudomonas aeruginosa ATCC 9027 *Bacillus subtilis ATCC 6633

*According European Pharmacopoeia in paragraph Water for Injections (Aqua ad iniectabilia) Inoculate 100 CFU at 30-35C 3 days.

BIBLIOGRAPHY
American Public Health Association (1985) Standard Method for the Enumeration of Water and Wasterwater. European Pharmacopeia 7.0.

EP
STORAGE
Once opened keep powdered medium closed to avoid hydration.
2C 25C

LABORATORIOS CONDA, S.A.

www.condalab.com