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• Oxoid Clostridium Difficile Selective Supplement is added to Clostridium Difficile Agar Base, Part No. CM0601B.
• Each vial supplements 500mL of medium.

Clostridium Difficile Selective Supplement, Oxoid Composition

Clostridium Difficile Agar, Oxoid Preparation:
Suspend 34.5g in 500ml of distilled water and bring gently to the boil to dissolve completely. Sterilise by autoclaving at 121°C for 15 minutes. Allow to cool to 50°C and add aseptically the contents of 1 vial of Oxoid Clostridium Difficile Supplement (SR0096) reconstituted as directed, together with 7% (v/v) Defibrinated Horse Blood (SR0050). Sheep Blood (SR0051) may be used in place of Horse Blood but some strains of the organism will show a slightly reduced growth recovery. Mix well and pour into sterile Petri dishes.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared medium at 2-8°C no longer than 5-7 days.

• Oxoid Clostridium Difficile Agar Base is used for the isolation of Clostridium difficile.
• Also available Clostridium Difficile Selective Supplement, Part No. SR0096E.

lostridium Difficile Agar Base, Oxoid Composition

Clostridium Difficile Agar, Oxoid Preparation:
Suspend 34.5g in 500ml of distilled water and bring gently to the boil to dissolve completely. Sterilise by autoclaving at 121°C for 15 minutes. Allow to cool to 50°C and add aseptically the contents of 1 vial of Oxoid Clostridium Difficile Supplement (SR0096) reconstituted as directed, together with 7% (v/v) Defibrinated Horse Blood (SR0050). Sheep Blood (SR0051) may be used in place of Horse Blood but some strains of the organism will show a slightly reduced growth recovery. Mix well and pour into sterile Petri dishes.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared medium at 2-8°C no longer than 5-7 days.

Precautions
Colonies of Clostridium difficile from faecal cultures are smaller when egg yolk is used in place of horse blood.

The Oxoid formula does not contain the neutral red indicator proposed by George et al. because it is designed for use with horse blood. On this medium the typical colour of the colony of Clostridium difficile will not appear however there will be a fluorescent reaction.

Typical Gram stain morphology of Clostridium difficile may not be evident in colonies picked from this medium because of the antibiotics present. Subculture to blood agar to obtain characteristic morphology.

Cultivate and isolate fastidious microorganisms with clearly visible hemolytic reactions with Thermo Scientific™ Oxoid™ Columbia Blood Agar Base (Dehydrated) when used with whole defibrinated blood (SR0050). Columbia Blood Agar Base is an ideal base medium for preparation of blood agar, chocolate agar and for preparation of various selective and identification media.

Columbia Blood Agar Base, Oxoid Composition

Columbia Blood Agar, Oxoid Preparation:
Add 39g to 1 litre of distilled water. Boil to dissolve the medium completely. Sterilise by autoclaving at 121°C for 15 minutes. Cool to 50°C and add 5% sterile defibrinated blood.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared plates of medium at 2-8°C.

Precautions
Brucella cultures are highly infective and must be handled under properly protected conditions. Incubate in 5-10% carbon dioxide atmosphere for 24-48 hours.
Campylobacter species are best grown at 42°C (except Campylobacter fetus subsp. fetus) in a micro-aerophilic atmosphere (Oxoid Campylobacter Gas Generating Kit BR0056 or BR0060 or CampyGen CN0025/CN0035).
Staph./Strep. supplemented plates should be incubated aerobically at 35°C for 18 hours. Incubation in carbon dioxide- enriched air will cause inhibition of staphylococcal growth.
Streptococcus Selective Supplement SR0126 supplemented plates may be incubated aerobically or anaerobically at 35°C for 18 hours.
Prepared plates of both supplemented media should be used within 18 hours of preparation for maximum selectivity. Gardnerella supplemented plates should be incubated at 35°C for 48 hours in an atmosphere containing 7% carbon dioxide.
Carry out confirmatory tests on all colonies from horse blood medium and on beta-haemolytic colonies from human or rabbit blood medium.
Incubate plates of Clostridium E-Y Agar anaerobically at 35°C for 18 hours, look for lecithinase activity (pearly layer) and for proteolysis. Lecithinase activity is inhibited in the presence of specific anti-toxin.