C.D.C. ANAEROBE 5% SHEEP BLOOD AGAR

Technical Data #1956a / 98.06.16

C.D.C. Anaerobe 5% Blood Agar is an enriched nonselective solid medium used for cultivation and isolation of obligate and facultative anaerobes from a clinical and non clinical specimens. Growth is enhanced by yeast extract, vitamin K and hemin.

FORMULA
in grams per liter purified filtered water

Casein Peptone	15,0
Soy Peptone	5,0
Yeast Extract	5,0
Sodium Chloride	5,0
Agar	15,0
Enrichments
Hemin	0,005
Vitamine K1	0,01
Sheep Blood	50 ml

pH 7,3 +/- 0,2 à 25º 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 2-8º C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25º 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 40 g of dehydrated media in 950 ml of purified filtered water. Add 5 g of Yeast Extract powder. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C and aseptically, add 50 ml of Sheep Blood (4245), 2x5 ml of Enrichment Vitamin K-Hemin (8753). Mix gently and dispense into sterile Petri dishes.

PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

1. Suitable anaerobes recovery is depending of collection and transportation of specimens. Specimens should be transferred into Anaerobes Pre-Reduced Transport Medium (2017) immediately after collection and brought to the laboratory quickly.
2. Directly, inoculate plate as soon as possible with a large amount of inoculum, streak for isolation.
3. Incubate at 35º C under anaerobic conditions for 2-4 days.
4. Examine for growth.

QUALITY CONTROL
Results after 48 hrs at 35ºC anaerobic atmosphere

Organisms	ATCC	Growth
Bacteroides fragilis	25285	+
Clostridium perfringens	13124	+ hemolysis
Fusobacterium nucleatum	25586	+
Bacteroides levii	29147	+
Peptostreptococcus anaerobius	27337	+

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

REFERENCES

1. Dowell, V.R. Jr., G.L. Lombard, F.S. Thompson, A.Y. Armfield 1977 Media for isolation, characterization, and identification of obligately anaerobic bacteria. CDC laboratory manual. Center for Disease Control, Atlanta.
2. Dowell, V.R. Jr., T.M. Hawkins 1979 Laboratory methods in anaerobic bacteriology. CDC laboratory manual. HEW Publication No. (CDC) 79-8272. Center for Disease Control, Atlanta.
3. Gibbons, R.J., J.B. Mac Donald 1960 Hemin and vitamin K compounds as required factors for the cultivation of certain strains of Bacteroides melaninogenicus. J. Bacteriol. 80:164-170.
4. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J 1985 Manual of Clinical Microbiology, 4th ed., Washington D.C.: American Society for Microbiology.
5. N.C.C.L.S. Quality Assurance for Commercially Prepared Microbiological Culture Media N.C.C.L.S. Publication M22-A Approved Standard 1990 Vol.10 No.14.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-5106
Yeast Extract	500 g	QB-39-5701
Prepared media
Plates, 100x15 mm	10/pkg	1956
Mono-Plates, 100x15 mm	10/pkg	1956M
Enrichment Vitamin
K-Hemin, 5ml	10/pkg	8753
Sheep Blood	50 ml	4245

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