BRUCELLA MEDIUM BASE
CAT Nº: 1374
For the cultivation of Brucella in diverse clinical specimens, foods and
other materials of sanitary interest
FORMULA IN g/l
Peptone
10.00
Sodium Chloride
5.00
Glucose
10.00
Bacteriological Agar
15.00
Beef Extract
5.00
Final pH 7.5 ± 0.2 at 25ºC
PREPARATION
Suspend 45 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 121°C for 15 minutes. Cool to 45- 50°C and add
5% horse serum, inactivated by heating at 56ºC for 80 minutes. To obtain a selective medium add 2 vials of Brucella
Selective Supplement Modified (Cat. 6060) previously reconstituted in 10 ml of 1:1 w/w of ethanol and distilled water.
Homogenize gently and pour into Petri dishes. The prepared medium should be stored at 8-14°C. The color is amber.
The dehydrated medium should be homogeneous, free-flowing and light in color. If there are any physical changes,
discard the medium.
Brucella Selective Supplement Modified (Cat. 6060 )
(1 vial for 500 ml of the medium)
Nystatin: ........................... 50000 UI
Bacitracin: ........................ 12500 UI
Polymyxin B: ...................... 2500 UI
Natamycin: ......................... 25.0 mg
Vancomycin: ...................... 10.0 mg
Nalidixic Acid: ...................... 2.5 mg
USES
BRUCELLA BASE MEDIUM is prepared according to the formula described by Jones and Brinley Morgan for the cultivation
and isolation of Brucella, including fastidious types. It is a medium rich in nutritive elements and growth factors that
make it adequate for the growth and isolation of Brucella spp.
Beef extract and Peptone provide nitrogen, vitamins, minerals and amino acids essential for growth. Glucose is the
fermentable carbohydrate providing carbon and energy. Sodium chloride supplies essential electrolytes for transport and
osmotic balance. Bacteriological agar is the solidifying agent. The addition of the supplement enhances the medium’s to
selectivity for the growth of Brucella.
Brucella species are level 3 pathogens and cause brucellosis, a zoonotic disease. It is usually transmitted through milk,
dairy products, meat and direct contact with infected animals.
It is widely used for the isolation of Brucella in highly contaminated materials, food materials and clinical samples.
Incubate at 35°C± 2°C in an atmosphere of 5-10% CO2 and observe alter 72 hours.
MICROBIOLOGICAL TEST
The following results were obtained in the performance of the medium, by adding the supplement, from type cultures
after incubation at a temperature of 35°C± 2°C, in an atmosphere of 5-10% CO2, and observed after 72 hours.
1
LABORATORIOS CONDA, S.A.
www.condalab.com
Microorganisms
Growth
Brucella abortus ATCC 4315
Good
Brucella melitensis ATCC 4309
Good
Brucella suis ATCC 4314
Good
BIBLIOGRAPHY
Kzudas and Mors, J.Bact. 66:502. 1953 Rennoux G. Ann. Inst. Pasteur, 87:325. 1954 Standard Methods for Examination of Diary
Products. 10 th Ed. APHA, Inc. New York 1960 Smith Louis Ds. The pathogenic anaerobic Bacteria. C. Thomas Pub. Springfield, II, 1975
STORAGE
25ºC
Once opened keep powdered medium closed to avoid hydration.
2ºC
2
LABORATORIOS CONDA, S.A.
www.condalab.com