BRUCELLA AGAR WITH HEMIN AND VITAMIN K (H & K)
|Cat. no. A30||Brucella Agar with H & K,15x100mm Plate, 19ml||10 plates/bag|
|Cat. no. H05||Brucella Agar with H & K, 15x150mm Plate, 69ml||10 plates/bag|
|Cat. no. J74||
Brucella Agar with H & K / Anaerobic PEA, 15x100mm Biplate,
|Cat. no. J87||
Brucella Agar with H & K / LKV Agar, 15x100mm Biplate,
Hardy Diagnostics Brucella Agar with H & K is recommended for use in the primary isolation and cultivation of anaerobic microorganisms.
Brucella Agar with H & K is modification of the formulation given by the American Society for Microbiology (ASM). (4) According to Finegold, this media is reported preferable to Heart Infusion Blood Agar Base for the cultivation of anaerobic bacteria. (2) Onderdonk, et al., and Weinstein, et al., both reported the addition of Hemin. (7,10) Summanen, et al., described supplementing the medium with vitamin K. (9) The incorporation of these additives, into Brucella Agar with H & K, enhances the cultivation of some species of anaerobes. (6)
Brucella Agar with H & K is a rich nutrient base that supports the growth of fastidious organisms. The media contains dextrose which provides an energy source; peptones to provide nitrogenous compounds, and yeast extract to supply B vitamins. Growth factors required by some anaerobic bacteria are provided for in the added sheep blood, which also allows the demonstration of hemolytic reactions. Hemin and vitamin K are incorporated into the medium to enhance the growth of gram-positive spore-formers and Bacteroides species. (6)
Ingredients per 950ml of deionized water:*
|Pancreatic Digest of Casein||10.0gm|
|Pancreatic Digest of Animal Tissue||10.0gm|
|Sheep Blood, Defibrinated||50.0ml|
Final pH 7.0 +/- 0.3 at 25ºC.
* Adjusted and/or supplemented as required to meet performance criteria.
STORAGE AND SHELF LIFE
Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.
Specimen Collection: Consult listed references for information on specimen collection. (1-5) Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.
When possible the clinical specimen should be inoculated directly onto the medium to prevent loss of organism viability. A liquid specimen may be directly applied to the agar surface and streaked with a sterile inoculating loop. Specimens for anaerobic culture should be plated on both non-selective and selective media.
Method of Use: Consult listed references for the correct inoculation procedure. (1-5,9) Incubate plates anaerobically at 35-37ºC. for up to 72 hours. Confirmation of anaerobic organisms should be performed by subculturing to an aerobic Blood Agar plate (Cat. no. A10). Examine anaerobic colonies for hemolytic reaction, colony morphology, gram stain, and further biochemical testing.
INTERPRETATION OF RESULTS
Consult listed references for the interpretation of growth of anaerobic species. (1-5,9)
It is suggested that Brucella Agar with H & K plates be reduced, prior to use, for a minimum of 24 hours by placing them in an anaerobe jar at room temperature.
MATERIALS REQUIRED BUT NOT PROVIDED
Standard microbiological supplies and equipment such as loops, slides, anaerobe jars or bags, anaerobic generators and indicators, incinerators, microscopes, and incubators, etc., as well as serological and biochemical reagents, are not provided.
|Test Organisms||Inoculation Method*||Incubation||Results|
ATCC ® 25285
ATCC ® 29147
ATCC ® 13124
ATCC ® 25586
ATCC ® 27337
User Quality Control
Brucella Agar with H & K should appear opaque, and cherry red in color.
Fusobacterium nucleatum (ATCC ® 25586) colonies growing on Brucella Agar with H and K (Cat. no. A30). Incubated anaerobically for 24 hours at 35ºC.
Bacteroides fragilis (ATCC ® 25285) colonies growing on Brucella Agar with H and K (Cat. no. A30). Incubated anaerobically for 24 hours at 35ºC.
Bacteroides levii (ATCC ® 27337) colonies growing on Brucella Agar with H and K (Cat. no. A30) Incubated anaerobically for 24 hours at 35ºC.
Clostridium perfringens (ATCC ® 13124) colonies growing on Brucella Agar with H and K (Cat. no. A30). Incubated anaerobically for 48 hours at 35ºC.
Peptostreptococcus anaerobius (ATCC ® 27377) colonies growing on Brucella Agar with H and K (Cat. no. A30). Incubated anaerobically for 48 hours at 35ºC.
Uninoculated plate of Brucella Agar with H and K (Cat. no. A30).
1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.
2. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.
3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.
4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.
5. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.
6. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams & Wilkins, Baltimore, MD.
7. Onderdonk, A.B., et al. 1974. Infect. Immun.; 10:1256.
8. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.
9. Summanen, P., et al. 1993. Wadsworth Anaerobic Bacteriology Manual, 5th ed. Star Publishing Company, Belmont, CA.
10. Weinstein, W.M., et al. 1974. Infect. Immun.; 10:1250.
ATCC is a registered trademark of the American Type Culture Collection.