Presence of Clostridium difficile PCR ribotype clusters related to 033, 078 and 045 in diarrhoeic calves in Germany.

This study provides data on the distribution and relationship of C. difficile FOR ribotypes in diarrhoeic calves in Germany. C. difficile was isolated from 176 of 999 (17.6%) faecal samples or swabs of diarrhoeic calves from 603 farms collected between January 2010 and August 201 2 by eight federal laboratories of six states. Strains were assigned to 17 PCR ribotypes. FOR ribotypes 033 (57%), 078 (17 %) and 045/FLI01 (closest match to 045 in the WEBRIBO database; 9%) were found the most frequently. Nine per cent of all culture-positive tested animals shed more than one multiple locus variable number tandem repeat analysis (MLVA) or PCR ribotype. Eight PCR ribotypes with related profiles (including 033, 078 and 045/FLI01) representing 92% of all isolates were grouped into three clusters. Molecular relatedness was supported by the absence of the MLVA locus A6(Cd) only in clustered strains and identical toxin gene profiles for strains within each cluster. Previously reported mulitilocus sequence typing analysis for PCR ribotypes that were also recovered in this study found identical sequence types and a tcdC deletion (Delta 39 bp) for 033, 045, 078 and 126 (ST-11), confirming this clustering. A different geographical occurrence of PCR ribotypes was shown for cluster 033 (found more frequently in southern Germany) and 045 (found more frequently in northern Germany). This study showed that clusters of C. difficile PCR ribotypes related to 033, 078 and 045 are predominant in diarrhoeic calves in Germany. The high number of strains belonging to PCR ribotype 078 demonstrated that diarrhoeic calves are also potential reservoirs for human pathogenic C. difficile strains.