The agents of brucellosis, Brucella species are normal flora of the genital and urinary tracts of many animals including goats, pigs, cows and dogs. Most humans acquire the disease through ingestion of contaminating milk or through occupational exposure; the disease is particularly common among abattoir workers (1).
Brucella Agar Base w/ Hemin and Vitamin K1 is a modified (4, 5, 6) and highly enriched medium, which can be used for the isolation of Brucella and other anaerobic bacteria (2, 3).
The medium contain casein enzymic hydrolysate, peptic digest of animal tissue and yeast extract as sources of carbon, nitrogen and essential growth nutrients including B-complex vitamins. Dextrose serves as a source of energy. Addition of blood provides nutrients and helps to differentiate hemolytic organisms (2, 3). Presence of hemin and Vitamin K1 supports growth of other fastidious bacteria like Bacteroides species and gram-positive spore bearers like Clostridium species (7). The specimen should be inoculated onto the plate (reduced earlier by placing under anaerobic conditions for 18- 24 hrs) as early as possible. Swab cultures are directly streaked. Non-swab cultures are inoculated using an inoculating loop.
Incubation is carried out anaerobically at 35°C for at least 48 hours; however, negative results should be reported only after incubation for 7 days.
Storage and Shelf-life:
Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.
References:
1. Baron E. J., Finegold S. M., (Eds.), 1990, Bailey and Scotts Diagnostic Microbiology, 8th Ed., The C.V. Mosby Co., St. Louis.
2. Zennette, Balows, Hausler and Shadomy, (Eds.), 1985, Manual of Clinical Microbiology, 4th Ed., ASM, Washington, D.C.
3. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams and Wilkins, Baltimore
4. Sutter V. L., Citron D. M. and Finegold S. M., 1985, Wadsworth Anaerobic Bacteriology Manual, 4th Ed., Star Publishing Co., Belmont, Ca.
5. Onderdonk A. B., Weinstein W. M., Sullivan N. M. and Bartlett J. G., 1974, Infect. Immun., 10:1256.
6. Weinstein W. M., Onderdonk A. B., Bartlett J. G. and Gorbach S. L., 1974, Infect. Immun., 10:1250.
7. Gibbons and MacDonald, 1960, J. Bacteriol., 80:164.