The study describes a novel TMrtPCR for the direct detection of C. difficile in animal faecal samples. TMrtPCR targets the genes for toxins A and B, and binary toxin; detection of the latter is becoming more important due to its potential clinical significance and association with CA-CDI. TMrtPCR could be the preferred screening method, because of its advantages: it is faster and it provides the first information about the toxin type. Because of possible false negative TMrtPCR results, enrichment culture should be performed for the specimens with negative or inconclusive rtPCR results as a second step for C. difficile detection in animal samples.
As migrating birds can be a key epizootiological factor for transmission and distribution of pathogens over a wide geographic range, a possible role of passerine birds in the spread of Clostridium difficile along the bird south-migration over the Alps was studied. Analysis of the cloacal samples from 465 passerine birds by selective enrichment generated negative results, indicating that migrating passerine birds are unlikely to serve as a reservoir and a carrier of C. difficile.
To evaluate the occurrence of Clostridium difficile in seafood and coastal environment samples (seawater, sediment, zooplankton and mussels were collected in the Gulf of Naples. Clostridium difficile was isolated from 9 (43%) of the 21 samples investigated and zooplankton was sample type with highest prevalence of positive samples. No C. difficile was detected in marine sediment. Five known different PCR-ribotypes ( 003, 009, 010, 056 and 066) were recognized in the C. difficile isolates and 50% of the C. difficile isolates were toxin A/B positive.