Seasonality of Clostridium difficile in the natural environment

Clostridium difficile is considered the leading cause of antibiotic-associated disease worldwide. In the past decade, a large number of studies have focused on identifying the main sources of contamination in order to elucidate the complete life cycle of the infection. Hospitals, animals and retail foods have been considered as potential vectors. However, the prevalence of C. difficile in these types of samples was found to be rather low, suggesting that other contamination routes must exist. This study explores the presence of C. difficile in the natural environment and the seasonal dynamics of the bacterium. C. difficile was isolated from a total of 45 samples out of 112 collected (40.2%) on 56 sampling points. A total of 17 points were positive only during the winter sampling (30.4%), 10 were positive only during the summer sampling (17.9%) and 9 sampling points (16.1%) were positive in both summer sampling and winter sampling. Spore counts in soil samples ranged between 50 and 250 cfu/g for 24.4% of the positive samples, with the highest concentrations detected in samples collected in the forest during winter campaign (200-250 cfu/g). A total of 17 different PCR ribotypes were identified, and 15 of them had the genes coding for toxins A and B. Most of those ribotypes had not previously been found or had been isolated only sporadically (<1% of samples) from hospitals in Belgium. Regarding antimicrobial susceptibility, most of the resistant strains were found during the summer campaign. These findings bear out that C. difficile is present in the natural environment, where the bacterium undergoes seasonal variations.