The caecal content (2 g of solid or 2 ml in case of liquid content)
was collected and stored at - 20 C until one gram of each sample
was added to 9 mL of C. difficile selective medium Taurocholate
Cefoxitine Cycloserine Fructose Broth (TCCFB, Oxoid) and incubated
at 37 C for ten days in anaerobic conditions. After incubation,
the broth cultures were vortexed and alcohol shocked for 1 hour at
room temperature after mixing 2 mL of broth with 96% ethanol
(1:1 v/v). The samples were subsequently centrifuged at 3800 xg
for 10 min and the sediment was streaked onto Taurocholate
Cefoxitine Cycloserine Fructose agar (TCCFA, Oxoid) and incubated
for 48-72 hours in anaerobic chamber. Presumptive identification