CRITERION™ COLUMBIA AGAR BASE

Cat. no. C5450 CRITERION™ Columbia Agar Base 85gm
Cat. no. C5451 CRITERION™ Columbia Agar Base 500gm
Cat. no. C5452 CRITERION™ Columbia Agar Base 2kg
Cat. no. C5453 CRITERION™ Columbia Agar Base 10kg
Cat. no. C5454 CRITERION™ Columbia Agar Base 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Columbia Agar Base is used for cultivating fastidious and non-fastidious microorganisms with or without the addition of blood. It is an enriched agar medium.

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

Columbia Agar Base media can be prepared as Blood Agar or Chocolate Agar. It is typically supplemented with 5-10 % sheep, rabbit or horse blood. Columbia Blood Agar is used for isolating, cultivating and determining the hemolytic reactions of fastidious pathogenic microorganisms. Without enrichment, Columbia Agar Base can be used as a general purpose medium.

Columbia Agar Base media employs specially selected raw materials to support growth of fastidious microorganisms. Columbia Agar Base supplemented with 5 or 10% sheep blood derives its superior growth-supporting properties from the combination of peptones prepared from pancreatic digest of casein, peptic digest of animal tissue and beef extract. Yeast extract and corn starch are also included in the formulation and serve as energy sources with yeast extract supplying B-complex vitamins. Columbia Agar Base has a relatively high carbohydrate content and therefore beta-hemolytic streptococci may produce a greenish hemolytic reaction (on media containing blood) that may be mistaken for alpha-hemolysis.

FORMULA

Gram weight per liter: 40.0gm/L
Pancreatic Digest of Casein 12.0gm
Peptic Digest of Animal Tissue 5.0gm
Sodium Chloride 5.0gm
Yeast Extract 3.0gm
Beef Extract 3.0gm
Corn Starch 1.0gm
Agar 11.0gm

Final pH 7.3 +/- 0.2 at 25ºC.

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

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 40.0gm 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. and aseptically add enrichments, if desired. Blood is generally added at a concentration of 5-10%.

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. A16.

LIMITATIONS

Columbia Agar Base is intended for use with blood supplementation. Although certain diagnostic tests may be performed directly on these media, biochemical and, if indicated, immunological testing using pure cultures is recommended for complete identification.

Since the nutritional requirements of organisms vary, some strains may be encountered that fail to grow or grow poorly on this medium.

Hemolytic reactions of some strains of group D streptococci have been shown to be affected by differences in animal blood. Such strains are beta-hemolytic in horse, human and rabbit blood agar and alpha-hemolytic on sheep blood agar.

Colonies of Haemophilus haemolyticus are beta-hemolytic on horse and rabbit blood agar and must be distinguished from colonies of beta-hemolytic streptococci using other criteria. The use of sheep blood has been suggested to eliminate this problem since sheep blood is deficient in pyridine nucleotides and does not support growth on H. haemolyticus.

The atmosphere of incubation has been shown to influenced hemolytic reactions of beta-hemolytic streptococci. For optimal performance, incubate blood agar media under increased CO2 anaerobic conditions.

Columbia Agar has a relatively high carbohydrate content and therefore beta-hemolytic streptococci may produce a greenish hemolytic reaction (on media containing blood) that may be mistaken for alpha-hemolysis.

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
With addition of blood:
Streptococcus pyogenes
ATCC ® 19615
A 18-48hr 35°C CO 2 ** Growth; beta-hemolysis
Streptococcus pneumoniae
ATCC ® 6305
A 18-48hr 35°C CO 2 ** Growth; alpha-hemolysis
Staphylococcus aureus
ATCC ® 25923
A 18-48hr 35°C CO 2 ** Growth; beta-hemolysis
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C CO 2 ** Growth
Peptostreptococcus anaerobius
ATCC ® 27337
A 24-48hr 35°C Anaerobic Growth
Without addition of blood:
Clostridium sporogenes ***
ATCC ® 19404
J 48hr 35°C Anaerobic Growth
Clostridium perfringens
ATCC ® 13124
J 48hr 35°C Anaerobic Growth

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

*** Tested in accordance with USP <61> and <62>.(6,7)

User Quality Control

PHYSICAL APPEARANCE

CRITERION™ Columbia Agar Base powder should appear homogeneous, free-flowing, and beige in color. The prepared medium should appear opalescent to opaque, and beige 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. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

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

6. The Official Compendia of Standards. USP General Chapter <61> Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests. USP-NF. United States Pharmacopeial Convention Inc., Rockville, MD.

7. The Official Compendia of Standards. USP General Chapter <62> Microbiological Examination of Nonsterile Products: Tests for Specified Microorganisms. USP-NF. United States Pharmacopeial Convention Inc., Rockville, MD.


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

051616gr