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

Technical Data #1934a / 2013.07.04

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 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 35C under anaerobic atmosphere

Organisms

ATCC

Growth

Bacteroides fragilis

25285

+

Clostridium perfringens

13124

+ 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

 


 





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