Campylobacter Blood agar

Cat. no. A137 Campylobacter Blood Agar, 15x100mm plate, 18ml 10 plates/bag

INTENDED USE

Hardy Diagnostics Campylobacter Blood Agar is recommended for the primary isolation and cultivation of Campylobacter jejuni from human fecal specimens. (1-4)

SUMMARY

Dekeyser et al. reported the isolation of Campylobacter jejuni from the feces of patients with diarrhea and acute gastroenteritis by using a filtration technique and a selective medium with antimicrobials to suppress the normal enteric flora. (5) In 1977, Skirrow reported a selective medium containing three antimicrobics. (6) Blaser et al. in 1978, reported success in isolating C. jejuni with a medium containing four antimicrobics incorporated into Brucella Agar supplemented with 10% defibrinated sheep blood. (1,2)

Peptones, dextrose, yeast extract, and blood support the growth of Campylobacter species. The peptones supply the nitrogen compounds, carbon, sulfur, and trace ingredients. Yeast extract is a source of B vitamins while dextrose is utilized as an energy source. Sheep blood supplies additional nutrients. The microbial agents, polymyxin B, trimethoprim, nystatin, and vancomycin, suppress the growth of the normal microbial flora in fecal specimens, thereby facilitating isolation of C. jejuni . Vancomycin inhibits gram-positive bacteria and polymyxin B inhibits most gram-negative bacilli except for Proteus . Trimethoprim is inhibitory for Proteus spp. Nystatin is an antifungal agent.

FORMULA

Ingredients per liter of deionized water:*

Agar 13.0gm
Trimethoprim 15.0gm
Pancreatic Digest of Casein 10.0gm
Pancreatic Digest of Animal Tissue 10.0gm
Sodium Chloride 5.0gm
Yeast Extract 3.0gm
Dextrose 1.0gm
Vancomycin 1.0gm
Nystatin 0.63gm
Polymyxin B 0.5gm
Sodium Bisulfite 0.1gm
Sheep Blood 100.0ml

Final pH 7.4 +/- 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

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection and handling. (8-11) Specimens should be obtained before antimicrobial therapy has been administered. 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, such as Campy Thioglycollate (Cat. no. K128) and refrigerate 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.

Incubate inoculated plates at 42ºC. in an atmosphere conducive to the primary isolation and cultivation of microaerophilic organisms such as Cat. no. CN020C, or CN025A. Examine plates at 24 and 48 hours.

INTERPRETATION OF RESULTS

Campylobacter jejuni will appear as small, mucoid colonies, usually grayish in coloration, flat with irregular edges, and nonhemolytic at 24-48 hours. (8) An alternate colonial morphology that appears to be strain related consists of round colonies 1-2mm in diameter that are convex, entire, and glistening. (8) Colonies tend to spread or swarm, especially when initially isolated from fresh clinical specimens.

If the plates are to be examined at 24 hours, treat as if they were anaerobic cultures. Examine the plates quickly and place them back into a reduced oxygen atmosphere immediately after examination.

Consult listed references for further interpretation of growth. (8-11)

LIMITATIONS

C. jejuni is thermophilic so it is important to incubate the plates at 42ºC. If incubated at lower temperatures growth may be delayed and the selectivity of the medium is reduced.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, anaerobe jars or bags, microaerophilic generators, 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
Campylobacter jejuni
ATCC ® 33291***
A 48hr 35°C Micro** Growth
Escherichia coli
ATCC ® 25922***
B 48hr 35°C Aerobic Partial to complete inhibition

** Microaerophillic atmospheric requirements, incubate in a jar with an appropriate atmospheric generator (Cat. no. CN025A).

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

USER QUALITY CONTROL

Physical Appearance

Campylobacter Blood Agar should appear opaque, and cherry red in color with no precipitate, chips or debris.

REFERENCES

1. Blaser, M., J. Cravens, B.W. Powers, and W.L. Wang. 1978. Campylobacter enteritis associated with canine infection. Lancet ii: 979-980.

2. Blaser, M.J., I.V. Berkowitz, F.M. LaForce, J. Cravens, L.B. Reller, and W-L. L. Wang. 1979. Campylobacter enteritis : clinical and epidemiologic features. Ann. Intern. Med. 91: 179-185.

3. Reller, L.B., W-L.L. Wang, and M.J. Blaser. 1979. Campylobacter enteritis : Campylobacter fetus subspecies jejuni . ASPC Check Sample, Microbiology No. MB-99, Commission on Continuing Education, American Society of Clinical Pathologists, Chicago.

4. Smith, J.P., K. Durfee, and J.H. Marymont, Jr. 1980. Incidence of Campylobacter enteritis in the midwestern United States. Am. J. Med. Technol. ; 46:81-84.

5. Dekeyser, P., M. Gossuin-Detrain, J.P. Butzler, and J. Sternon. 1972. Acute enteritis due to related Vibrio : first positive stool cultures. J. infect. Dis. ; 125:390-392.

6. Skirrow, M.B. 1977. Campylobacter enteritis : a "new" disease. Br. Med. J. 2: 9-11.

7. 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.

8. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

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

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

11. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

12. 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.


040116sw