CRITERION™ CHARCOAL AGAR

Cat. no. C7600 CRITERION™ Charcoal Agar 133gm
Cat. no. C7601 CRITERION™ Charcoal Agar 500gm
Cat. no. C7602 CRITERION™ Charcoal Agar 2kg
Cat. no. C7603 CRITERION™ Charcoal Agar 10kg
Cat. no. C7604 CRITERION™ Charcoal Agar 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Charcoal Agar is recommended for the cultivation and isolation of Bordetella pertussis.

SUMMARY

Charcoal Agar is formulated according to the method described by Mishulow, Sharpe and Cohen. They found Charcoal Agar to be an efficient substitute for the Bordet-Gengou Agar in the production of B. pertussis vaccines.(3)

The four Bordetella species, B. pertussis, B. parapertussis, B. bronchiseptica, and B. avium, are all respiratory pathogens that reside on the mucous membranes of the respiratory tract.(4) B. pertussis is the major cause of whooping cough or pertussis. B. parapertussis is associated with a milder form of whooping cough.(4) B. bronchiseptica is an opportunistic human pathogen, often occurring in patients having close contact with animals.(4) B. avium has not been reported in humans.

CRITERION™ Charcoal Agar contains beef extract, meat peptone, proteose peptone, and yeast extract as sources of nutrients. Sodium chloride is added to maintain osmotic balance and the starch is added to absorb toxic by-products. Nicotinic acid and charcoal provides selective properties and growth requirements.

Charcoal Agar can be supplemented with horse blood for use in the cultivation and isolation of Haemophilus influenzae.(2)

FORMULA

Gram weight per liter: 66.5gm/L
Beef Extract 10.0gm
Peptic Digest of Animal Tissue 10.0gm
Proteose Peptone 10.0gm
Starch 10.0gm
Yeast Extract 5.0gm
Sodium Chloride 5.0gm
Charcoal 4.0gm
Nicotinic Acid 0.001gm
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 gray.

Store the prepared media in plates at 2-8ºC.

PRECAUTIONS

METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 66.5gm 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. Mix thoroughly during dispensing to uniformly distribute the charcoal.

PROCEDURE

Specimen Collection: 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. Consult listed references for information regarding the processing and inoculation of specimens.(2-6,9)

The medium should be brought to room temperature prior to inoculation. Prepared media should be inoculated as to produce isolated colonies. Incubate aerobically in a humidified atmosphere with CO2 at 35ºC. Incubate up to 96 hours and examine daily for growth.

INTERPRETATION OF RESULTS

Refer to appropriate references and procedures for results.(1-4)

LIMITATIONS

Charcoal has a tendency to settle out of the medium. Swirl the flask gently when dispensing to obtain a uniform charcoal suspension.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as autoclaves, incinerators, and incubators, etc., are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Bordetella pertussis
ATCC ® 9340
A 48-96hr 35°C CO 2 ** Growth
Bordetella parapertussis
ATCC ® 15311
A 18-72hr 35°C Aerobic Growth

** Atmosphere of incubation is enriched with 5-10% CO2.

User Quality Control

PHYSICAL APPEARANCE

CRITERION™ Charcoal Agar powder should appear homogeneous, free-flowing, and gray in color. The prepared media should appear opaque with no chips or debris, and black in color.

REFERENCES

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

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

3. Mishulow, L., L.S. Sharpe and L.L. Cohen. 1953. Beef-heart charcoal agar for the preparation of pertussis vaccines. Am. J. Public Health; 43:1466.

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


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

051616gr