COLUMBIA BLOOD AGAR

Cat. no. A16 Columbia Blood Agar, 15x100mm Plate, 17ml 10 plates/bag
Cat. no. A16BX Columbia Blood Agar, 15x100mm Plate, 17ml 100 plates/box

INTENDED USE

Hardy Diagnostics Columbia Blood Agar is recommended for use as a general growth medium for the cultivation and hemolytic determination of fastidious and non-fastidious microorganisms from clinical specimens.

SUMMARY

Columbia Blood Agar was first described in 1966 by Ellner, Stoessel, Drakeford, and Vasi who incorporated animal derived peptone, enzymatic digests of casein, and defibrinated sheep blood into one medium. (3) It was found to be an improved blood agar, promoting both luxuriant and rapid growth, improved pigment production, typical colony morphology, and sharply defined hemolytic reactions.

REAGENT FORMULA

Ingredients per liter of deionized water:*

Enzymatic Digests of Casein 10.0gm
Casein Yeast Peptone 10.0gm
Sodium Chloride 5.0gm
Tryptic Digest of Beef Heart 3.0gm
Corn Starch 1.0gm
Sheep Blood 50.0ml
Agar 15.0gm

Final pH 7.3 +/- 0.2 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.

PRECAUTIONS

Specimen Collection: Consult listed references for information on specimen collection. (1,2,4,6) 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.

Method of Use: Allow the plates to warm to room temperature, and the agar surface to dry before inoculating. Inoculate and streak the specimen as soon as possible after collection. If the specimen to be cultured is on a swab, roll the swab over a small area of the agar surface. Streak for isolation with a sterile loop. Incubate plates aerobically or in a 5% to 10% carbon dioxide atmosphere at 35-37ºC. for 18-24 hours. Examine colonial morphology, characteristics, and hemolytic reactions.

INTERPRETATION OF RESULTS

Consult listed references for the identification of colony morphology and further biochemical tests required for identification. (1,2,4,6)

LIMITATIONS

Some beta-hemolytic streptococci strains will develop green hemolytic zones on Columbia Blood Agar. It is recommended to subculture all such streptococci strains onto Blood Agar plates (Cat. no. A10) to verify this reaction.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, 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
Streptococcus pyogenes
ATCC ® 19615
A 24hr 35°C Aerobic 2 ** Growth, beta-hemolysis
Streptococcus pneumoniae
ATCC ® 6305
A 24hr 35°C Aerobic 2 ** Growth, alpha-hemolysis
Staphylococcus aureus
ATCC ® 25923
A 24hr 35°C Aerobic 2 ** Growth
Escherichia coli
ATCC ® 25922
A 24hr 35°C Aerobic 2 ** Growth

User Quality Control

PHYSICAL APPEARANCE

Columbia Blood Agar should appear opaque, and cherry red in color.

S. pyogenes growing on Columbia Blood Agar

Streptococcus pyogenes (ATCC ® 19615) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.

S. pneumoniae growing on Columbia Blood Agar

Streptococcus pneumoniae (ATCC ® 6305) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.

S. aureus growing on Columbia Blood Agar

Staphylococcus aureus (ATCC ® 25923) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.

E. coli growing on Columbia Blood Agar

Escherichia coli (ATCC ® 25922) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.

Columbia Blood Agar

Uninoculated plate of Columbia Blood Agar (Cat. no. A16).



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.

5. Tille, P.M., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Ellner, et al. 1966. Am. Journ. Clin. Path. ; 45:502.

4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

5. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria , Vol. I. Williams & Wilkins, Baltimore, MD.

6. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

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

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

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