Cycloserine-Cefoxitin Fructose Agar with Lysozyme – CCFAL

Cycloserine-Cefoxitin Fructose Agar with Lysozyme – CCFAL

Cycloserine Cefoxitin Fructose Agar with Lysozyme (CCFA-L) is an enriched, selective, and differential media used for the isolation and presumptive identification of Clostridium difficile, a recognized cause of pseudomembranous (antimicrobial agent-associated) colitis.

CCFA-L Plate

C. difficile (ATCC 700057) Growing On CCFA-L Plate

1 plate per package

Catalog #
AS-2133 CCFA-L mono plate 1 plate
CCFA-L Product InsertCCFA-L SDS

Product Description & Summary

CCFA-L is an enriched, selective, and differential media for the isolation of Clostridium difficile. The basic nutritive base consists of animal peptones and fructose and is supplemented with cefoxitin and cycloserine at concentrations that inhibit the growth of most normal fecal flora. Cycloserine will inhibit gram-negative bacteria, while cefoxitin will inhibit both gram-positive and gram-negative organisms. Clostridium difficile is not inhibited on CCFA-L media, and when grown on this media, will exhibit a characteristic yellow, ground-glass colonial morphology. Neutral red is added as a pH indicator and lysozyme is added to aid in the germination of spores. The presence of C. difficile will turn the indicator from pink/orange to yellow, due to the utilization of amino acids by the organism causing an increase in the pH. This media is prepared, dispensed, and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

Interpretation of Results & Limitations

If used properly, CCFA-L supports good growth of members of C. difficile. After 24-48 hours, most colonies of C. difficile are large, circular, and yellow, with the yellow coloration extending 2 –3 mm beyond the colony into the initially pink/orange media. These colonies fluoresce golden yellow/chartreuse under long-wavelength UV light. Most other bacteria are inhibited on this media. At 48 hours, colonies of most other organisms (e.g. Lactobacilli and yeast), which may grow, are very small (pinpoint to 0.5 mm in diameter) and do not fluoresce golden-yellow.

CCFA-L will not provide complete information for identification of bacterial isolates. Rare strains of C. difficile may be inhibited.  Plates must be examined no later than after 48 hours of incubation for optimal selectivity. Significant numbers of colonies, other than C. difficile, may grow after 48 hours of incubation. A test for aerotolerance should be used to confirm that each colony type is an obligate anaerobe. Consult reference materials for additional information.

Quality Control

The following organisms are routinely used for quality control performance testing at Anaerobe Systems.

Organism Tested ATCC # Results Time Special Reaction
Bacteroides fragilis 25285 No Growth
Enterococcus faecalis 29212 No Growth
Clostridium sporogenes 3584 No Growth
Clostridium beijerinckii 8260 No Growth
Proteus mirabilis 12453 No Growth
Clostridium perfringens 13124 No Growth
Clostridium innocuum 14501 No Growth
Clostridium sordellii 9714 No Growth
Clostridium difficile 9689 Growth 24 hrs Yellow coloration, chartreuse fluorescence
Clostridium difficile 700057 Growth 24 hrs Yellow coloration, chartreuse fluorescence

Product Storage & Shelf Life

Storage: Upon receipt, store at 2 – 8oC in original container until used. Avoid overheating or freezing. Do not use media if there are signs of deterioration (shrinking, cracking, or discoloration due to oxidation of media) or contamination. The expiration date applies to the product in its original packaging and stored as directed. Do not use product past the expiration date shown on the container.

Shelf Life: 90 days from date of manufacture.