Brucella broth

Cat. no. R18 Brucella Broth, 13x100mm Tube, 1ml 20 tubes/box

INTENDED USE

Hardy Diagnostics Brucella Broth is recommended for use as a general purpose growth media for the cultivation of a variety of fastidious organisms, such as Streptococcus and Brucella spp. in clinical and food samples.

SUMMARY

Brucella broth is rich in nutrients and growth factors making it very suitable to grow and isolate a wide variety of microorganisms. This medium is used extensively for the cultivation of Brucella species along with the cultivation of many anaerobes from diagnostic specimens. Brucella Broth can also be used as the liquid medium component in blood culture bottles. (2)

Brucella Broth supports the growth of fastidious microorganisms with peptones, dextrose and yeast extract. The peptones provide nitrogen, vitamins, minerals and amino acids promoting growth. Yeast extract is a source of the
B-complex vitamins. Dextrose is an energy source as a fermentable carbohydrate. The sodium bisulfite is a reducing agent and sodium chloride maintains osmotic balance.

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digest of Casein 10.0gm
Peptic Digest of Animal Tissue 10.0gm
Sodium Chloride 5.0gm
Yeast Extract 2.0gm
Dextrose 1.0gm
Sodium Bisulfite 0.1gm

Final pH 7.0 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Cat. no. R18 should be stored at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration, discoloration , contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

Brucella spp. are classified as Biosafety Level 3 pathogens. Procedures with live cultures and antigens must be confined to a Class II biological safety cabinet (BSC). Biosafety Level 2 practices, containment equipment and facilities are recommended for routine clinical specimens of human or animal origin that contains or is believed to contain pathogenic Brucella spp. (2,11)

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection. (2-4,7-10) Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat, cold, and oxygen exposure. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport medium (Cat. no. S120D) and refrigerated until inoculation.

Method of Use: Consult the listed references for the appropriate cultivation techniques using this medium. (2-4,7-10)

1. The medium can be inoculated with a pure culture of an isolated colony, macerated tissue or liquid from a clinical specimen or a food sample.

2. For the cultivation of Brucella spp., incubate at 35ºC in duplicate. Incubate one set under aerobic conditions and another set at 5-10% CO 2.

3. For the cultivation of other organisms, incubate at 35 degrees with or without 5-10% CO 2 depending on the most suitable atmosphere to encourage growth.

INTERPRETATION OF RESULTS

Consult listed references for the interpretation of growth and other identification tests to identify growth of organism in this medium. (2-5, 8, 9 ,10)

The presence of growth is indicated by the appearance of turbidity.

LIMITATIONS

Brucella Broth is a general purpose growth media. Organisms growing in Brucella Broth will require further biochemical and/or serological testing for complete identification.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Bacteroides fragilis
ATCC ® 25285
A 24-72hr 35°C Anaerobic Growth
Clostridium perfringens
ATCC ® 13124
A 24-72hr 35°C Anaerobic Growth
Streptococcus pyogenes
ATCC ® 19615***
A 24-72hr 35°C CO 2 ** Growth

** Atmosphere of incubation is enriched with 5-10% CO 2 .

*** Recommended QC strains for User Quality Control according to the CLSI document M22 when applicable.

USER QUALITY CONTROL

Physical Appearance

Brucella Broth should appear clear and light amber in color, with no precipitates or debris.

REFERENCES

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. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

7. Association of Official Analytical Chemists. Official Methods of Analysissm, AOAC, Washington, D.C.

8. American Public Health Association. 1992. Standard Methods for the Examination of Dairy Products, 16th ed. APHA, Washington, D.C.

9. APHA Technical Committee on Microbiological Methods for Foods. 2001. Compendium of Methods for the Microbiological Examination of Foods, 4th ed. APHA, Washington, D.C.

10. U.S. Food and Drug Administration. Bacteriological Analytical Manual. AOAC, Arlington, VA.
http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm2006949.htm.

11. U.S. Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. 2007. Biosafety in microbiological and biomedical laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Government Printing Office, Washington, D.C.

ATCC is a registered trademark of the American Type Culture Collection.