Brucella Agar
Use and description: The medium is recommended for the cultivation of Brucella spp. and other fastidious organisms. The medium’s nutritive value is high due to its peptone, yeast extract and glucose content. Yeast extract is a source of vitamin B complex. The medium is supplemented with vitamin K1 (10 mg/l) and haemin (10 mg/l) to favour the growth of anaerobic bacteria. It can be supplemented with 5% blood to cultivate particularly fastidious aerobic and anaerobic bacteria and for the isolation of Campylobacter in microaerophil circumstances. To overcome the problem of slow growth of Brucella spp., the following antibiotics can be added to increase selectivity and suppress the growth of contaminant organisms: polymixin, vancomycin, bacitracin and cycloheximide and nystatin.
Composition per liter: Peptones………………………………………..18.0 g Yeast extract…………………………………….2.0 g Glucose………………………………………….1.0 g Sodium chloride………………………………...5.0 g Sodium bisulfite………………………………...0.1 g Vitamin K1………………………… …………0.01 g Hemin………………………… ………………0.01 g Agar……………………………………………15.0 g
Final pH of the ready to use medium: 7.0 ± 0.2
Medium preparation: Add 41.0 grams of dehydrated culture medium to 1 liter of distilled water. Heat with repeated stirring and boil for one minute to dissolve completely. Autoclave for 15 min at 121oC. To prepare blood agar plates or selective medium, cool medium to 48oC and add aseptically 5% sterile defibrinated blood and appropiate amount of Brucella selective supplement.
Quality specifications: Dehydrated medium: homogeneous, light beige fine powder. Ready to use medium: yellow beige and clear to slightly hazy, or cherry red, opaque if blood added
Microbiological response at 35 °C after 48-72 hours:
Storage: Dehydrated medium should be stored between 10 to 25oC. Once opened, place the container in a dark, dry place. The dehydrated medium should not be used if there is any lump or the color has changed from the original. |
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