CYCLOSERINE CEFOXITIN FRUCTOSE AGAR (C.C.F.A.)

Technical Data #1150a / 98.04.30

The C.C.F.A. Agar is a solid selective medium used for isolation and recovery of Clostridium difficile from feces without any pre-treatment. The pre-reduced treatment is not absolutely needed because of cystein HCl addition. Cycloserin and cefoxitin, the selective agents, prevent intestinal normal flora growth.

FORMULA
in grams per liter purified filtered water

Casein Peptone

10,0

Meat Peptone

10,0

Yeast Extract

3,0

Dextrose

1,0

Starch, soluble

1,0

Sodium Chloride

5,0

Sodium Bicarbonate

0,4

Sodium Pyruvate

1,0

Cystein HCI

0,5

L-Arginine

1,0

Pyrophosphate, soluble

0,25

Fructose

6,0

Succinic Acid

0,5

Hemin

0,01

Neutral Re

0,03

Agar

13,5

Supplements

 

Cycloserin

0,5

Cefoxitin

0,016

Egg Yolk, 50%, Sterile

50 ml

pH 7.2 +/- 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. Medium must not be used if there are signs of contamination or if the expiry date has passed.


PROCEDURE
Bring prepared media at room temperature before inoculation.

  1. Inoculate specimens without any pre-treatment. Streak to obtain isolated colonies.
  2. Incubate at 35º C under anaerobic atmosphere for 48 hours.


QUALITY CONTROL
Results after 48 hrs at 35ºC anaerobically

Organisms

ATCC

Growth

Colour

Clostridium difficile

9689

+

yellow

Escherichia coli

25922

-

 

Staphylococcus aureus

25923

-

 

Colonies of Clostridium difficile, are yellow, average of 4 mm- diameter, rounds with filamentous edges, low umbonate. Because of high selectivity of medium, it is recommanded to inoculate also a non-selective anaerobic blood agar and incubate under anaerobic atmosphere.


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


REFERENCES

  1. George, W.L., V.L. Sutter, D. Citron and S.M. Finegold 1979 Selective and differentiel medium for Isolation of Clostridium difficile. J. Clin. Microbiol. 9:214-219.
  2. 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.
  3. Mac Faddin, Jean F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria Vol.1 Baltimore, MD. Williams & Wilkins.


CATALOGUE NUMBERS

Prepared media    
Plates, 100x15 mm 10/pkg 1150
Mono-Plates, 100x15 mm 10/pkg 1150M
     
Egg Yolk, 50% sterile 50 ml 8653
Clostridium Supplement 10 x 5 ml vials 8705

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