C.D.C.-P.E.A. ANAEROBE 5% SHEEP BLOOD AGAR

Technical Data #1934a / 2000.07.04

C.D.C.-P.E.A. Anaerobe 5% Blood Agar is an enriched selective
and solid medium used for cultivation and isolation of anaerobes from a clinical and non clinical specimens. Yeast extract, vitamin K and hemin enhance anaerobes growth; phenylethanol inhibits gram-negative bacilli.

FORMULA
in grams per liter purified filtered water

Casein Peptone	15,0
Soy Peptone	5,0
Yeast Extract	5,0
Sodium Chloride	5,0
Phenylethanol	2,5
Agar	15,0
Enrichments
Hemin	0,005
Vitamine K1	0,01
Sheep Blood	50 ml

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 2-8° 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 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 121° C for 15 minutes. Cool to 45-50° 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 35° C under anaerobic conditions for 2-4 days.
4. Examine for growth.
   
   

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
Proteus mirabilis	12453	- 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 1985 Manual of Clinical Microbiology, 4th ed., Washington D.C.: American Society for Microbiology.
2. 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
3. Sutter, V.L., D. Citron, M.A.C. Edelstein and S.M. Finegold Wadsworth Anaerobic Bacteriology, 4th ed., C.V. Mosby, St Louis.
4. Wilkens, T.D., S.L. Chalgren, F. Jimenez-Ulate C.R. Drake Jr., J.L. Johnson 1976 Inhibition of Bacteroides fragilis on blood agar plates and reversal of inhibition by added hemin. J. Clin. Microbiol. 3:359-363

CATALOGUE NUMBERS

Dehydrated media	500 grams	QB-39-3521
Yeast Extract	500 grams	QB-39-5701
Prepared media
Plate 100x15 mm	10/pkg	1934
Mono-Plate 100x15 mm	10/pkg	1934M
Enrichment
Vitamin K-Hemin, 5ml	10/pkg	8753
Sheep Blood	50 ml	4245

2331, Dandurand	Phone	Fax	Internet
Montreal (Quebec)	(514) 277-2558	(514) 277-4714	http://www.quelab.com
Canada H2G 3C5	1 (800) 579-0998	1 (800) 579-8177	info@quelab.com