CRITERION™ CAMPYLOBACTER AGAR BASE

Cat. no. C5330 CRITERION™ Campylobacter Agar Base 74gm
Cat. no. C5331 CRITERION™ Campylobacter Agar Base 500gm
Cat. no. C5332 CRITERION™ Campylobacter Agar Base 2kg
Cat. no. C5333 CRITERION™ Campylobacter Agar Base 10kg
Cat. no. C5334 CRITERION™ Campylobacter Agar Base 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Campylobacter Agar Base is used for the selective isolation of Campylobacter species, particularly C. jejuni and C. coli, from clinical specimens and food samples. Campylobacter Agar Base is usually supplemented with sheep blood or lysed horse blood and selective agents such as Campylobacter Supplement by Oxoid - Skirrow formula, Cat. no. SR69E.

This dehydrated culture medium is a raw material intended to be used in the making of prepared media products, which will require further processing, additional ingredients, or supplements.

SUMMARY

Campylobacter species are microaerophilic organisms that inhabitant the gastrointestinal tracts of various animals, including poultry, dogs, cats, sheep, and cattle. C. jejuni and C. coli are the most common Campylobacter species associated with gastrointestinal infection. Symptoms usually include fever, abdominal cramping, and diarrhea lasting for several days to more than 1 week. Symptomatic infections, such as gastroenteritis, are usually self-limiting and do not require antibiotic therapy, although relapses may occur in 5 to 10% of untreated patients. Deaths attributed to C. jejuni infection are uncommon.(1,2,5,6)

Ingestion of improperly handled or undercooked food, primarily poultry products, raw milk, or contaminated water are common sources for human infections. The highest incidence of food borne illness occurs in infants and young children. Additionally, travelers to developing countries are also at risk for Campylobacter infections. (1,2,4-6) Unpasteurized milk has been a commonly implicated vehicle in foodborne outbreaks of C. jejuni enteritis. Survival of many strains of C. jejuni in raw milk is poor, and typically small numbers of the organism are present. This may explain the low incidence and difficulty in recovering this organism from suspect milk samples. In general, C. jejuni grows poorly in food and dies rapidly when exposed to ambient temperatures and atmospheres. However, it takes relatively few Campylobacter cells to cause illness and or symptoms of gastroenteritis in humans. (4,5,11)

CRITERION™ Campylobacter Agar Base contains proteose peptone no. 3 as a nitrogen source. Yeast Extract supplies B vitamins and growth stimulation. Beef extract provides a source of vitamins, carbon, nitrogen and amino acids. The addition of blood supplements the medium with X-factor and other growth requirements. The Campylobacter Supplement (Cat. no. SR69E) provides the selective agents to suppress the growth of normal fecal flora.

FORMULA

Gram weight per liter: 37.0gm/L
Proteose Peptone No.3 12.5gm
Yeast Extract 5.0gm
Sodium Chloride 5.0gm
Beef Extract 2.5gm
Agar 12.0gm

Final pH 7.4 +/- 0.2 at 25ºC.

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

STORAGE AND SHELF LIFE

Store the sealed bottle(s) containing dehydrated culture medium at 2-30ºC. Dehydrated culture medium is very hygroscopic. Keep lid tightly sealed. Protect dehydrated culture media from moisture and light. The dehydrated culture media should be discarded if it is not free-flowing or if the color has changed from its original beige.

Store the prepared culture media at 2-8ºC.

PRECAUTIONS

METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 37.0gm of the dehydrated culture media in 1 liter of distilled or deionized water.

2. Heat to boiling and mix to dissolve completely.

3. Sterilize in the autoclave at 121ºC. for 15 minutes.

4. Cool to 45-50ºC.

5. Aseptically add animal blood and selective agents (Cat. no. SR69E) as desired.

PROCEDURE AND INTERPRETATION OF RESULTS

For information on procedures and interpretation of results, consult listed references or refer to the prepared media Instructions for Use (IFU) for Cat. No. A40.

LIMITATIONS

Campylobacters are not easily visualized with the safranin counterstain normally used in the Gram stain procedure; therefore carbol fuchsin or 0.1% aqueous basic fuchsin can be used as the counterstain, or extending the staining time of the safranin to at least 10 minutes can improve the intensity of the stain.(1,6)

Most Campylobacter species require a microaerophilic atmosphere containing approximately 5% O2, 10% CO2, and 85% N2 for optimal recovery. The concentration of oxygen generated in candle jars is not optimal for the isolation of Campylobacter spp. and should not be used.(1)

Certain Campylobacter species, such as C. sputorum, C. concisus, C. mucosalis, etc., may require hydrogen for primary isolation and growth.(1)

The antimicrobial agents that are present in some Campylobacter Selective media, such as cephalothin, colistin, and polymyxin B, may be inhibitory to some strains of C. jejuni and C. coli, and are inhibitory to C. fetus. Furthermore, C. jejuni subsp. doylei and C. upsaliensis generally do not grow on cephalothin-containing media.(1)

Campylobacter Agar Base prepared with either Campylobacter Antimicrobic Supplement S or Campylobacter Supplement B is selective primarily for Campylobacter species. Biochemical testing using a pure culture is necessary for complete identification.

Some strains of normal enteric organisms may be encountered that are not inhibited or only partially inhibited on Campylobacter selective agents.

MATERIALS REQUIRED BUT NOT PROVIDED

Blood products and Campylobacter Supplements must be purchased separately. Standard microbiological supplies and equipment such as loops, other culture media, slides, staining reagents, pasteur pipettes, microaerophilic atmosphere packets, incubation jars, catalysts, 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 Agar Base with Supplement (Cat. no. SR69E):
Campylobacter jejuni
ATCC ® 33291
A 24-48hr 37°C Microaerophilic** Growth
Escherichia coli
ATCC ® 25922
B 24-48hr 37°C Aerobic Inhibited

** Atmosphere of incubation is enriched with 5% O2, 10% CO2 and 85% N2.

User Quality Control

PHYSICAL APPEARANCE:

CRITERION™ Campylobacter Agar Base powder should appear homogeneous, free-flowing, and beige in color. The prepared media, after the addition of animal blood, should appear opaque with no hemolysis, and cherry red in color.

REFERENCES

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

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

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

4. Marshall, R.T., ed. 1992. Standard Methods for the Examination of Dairy Products, 16th ed. APHA, Washington, D.C.

5. Vanderzant, C. and D.F. Splittstoesser, (ed.). 1992. Compendium of Methods for the Microbiological Examination of Foods, 3rd ed. APHA, Washington, D.C.

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

7. Isenberg, H.D. 1998. Essential Procedures for Clinical Microbiology. American Society for Microbiology, Washington, D.C.

8. Gun-Munro, J., R.P. Rennie, J.H. Thornley, H.L. Richardson, D. Hodge, and J. Lynch. 1987. Laboratory and clinical evaluation of isolation media for Campylobacter jejuni. J. Clin. Microbiol.; 25: 2274-2277.

9. Karmali, M.A., A.E. Simor, M. Roscoe, P.C. Fleming, S.S. Smith, and J. Lane. 1986. Evaluation of a blood-free, charcoal-based, selective medium for the isolation of Campylobacter organisms from feces. J. Clin. Microbiol.; 23:456-459.

10. Centers for Disease Control and Prevention. 2001. Campylobacter infections. www.cdc.gov/ncidod/dbmd/diseaseinfo/campylobacter_g.

11. Hunt, J.M., C. Abeyta and T. Tran. 1998. Chap. 7 Campylobacter. Bacteriological Analytical Manual, 8th ed., Rev. A. vm.cfsan.fda.gov/~ebam/bam-7.html.


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