COLUMBIA ANAEROBE GENTAMYCIN
5% SHEEP BLOOD AGAR

Technical Data #1923a / 2000.11.01

Columbia anaerobe Agar with gentamycin and 5% sheep blood is an enriched selective medium used for the isolation of obligately and facultatives anaerobes.
Gentamycin inhibits mixed normal flora.

FORMULA
in grams per liter purified filtered water

Casein Peptone	10,0
Meat Peptone	5,0
Yeast Extract	5,0
Beef Extract	3,0
Starch	1,0
Sodium Chloride	5,0
Agar	13,5
Hemin	0,005
Vitamin K	0,01
Defibrinated Sheep Blood	50,0 ml
Antibiotics
Gentamycin	0,14

pH 7,3 +/- 0,2 à 25° C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-8° C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25° C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 42,5 g in 950 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121° C for 15 minutes. Cool to 45-50° C and aseptically add 50 ml of Defibrinated Sheep Blood (4245), 2 tubes of Vitamin K-Hemin (8753) and a solution of 0,14 g sterile gentamycin. Dispense into sterile Petri dishes.


PROCEDURE
Reduce medium for 18-24 hours under anaerobic atmosphere before inoculation.

1. Inoculate plates, as soon as possible, with specimens and streak for isolation.
2. Very quickly, not more than 20 minutes, incubate at 35° C under anaerobic atmosphere for 48 hours.

QUALITY CONTROL
Results after 48 hrs at 35ºC under anaerobic atmosphere

Organisms	ATCC	Growth
Bacteroides fragilis	25285	+
Clostridium perfringens	13124	+ ß hemolysis
Fusobacterium nucleatum	25586	+
Bacteroides levii	29147	+
Peptostreptococcus anaerobius	27337	+
Escherichia coli	25922	- or partial
Staphylococcus aureus	25923	- or partial

LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.


REFERENCES

1. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American Society for Microbiology.
2. Mac Faddin, J.F. 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol.1 Williams and Wilkins, Baltimore.
3. N.C.C.L.S. Quality of Assurance for Commercially Prepared Microbiological Culture Media N.C.C.L.S. Publication M22-A Approved Standard 1990 Vol. 10 No. 14
4. Sutter, V.L., D. Citron, M.A.C. Edelstein and S.M. Finegold Wadsworth Anaerobic Bacteriology 4th, ed., C.V.Mosby,St. Louis

CATALOGUE NUMBERS

Dehydrated media	500 grams	QB-39-1006
Defibrinated Sheep Blood	50 ml	4245
Prepared media
Plate 100x15 mm	10/pkg	1923
Mono-Plate 100x15 mm	10/pkg	1923M
Vitamin K-Hemin, 5ml/tube/500 ml of medium	10/pkg	8753

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