Clostridium difficile (Clostridioides difficile) is an anaerobic, Gram-positive sporogenic bacterium and is a typical pathogen for humans and animals. The most important reservoir of the bacterium is the human and animal gut. But the spores can be detected in various environments (hospitals, water environments, sewage treatment plants, households, food). The aim of our study was to isolate C. difficile from sediments of various water environments in the area of eastern Slovenia and characterize strains with ribotyping and toxinotyping. Sediments were sampled at 15 different locations from six different types of surface waters. C. difficile spores were detected in all samples. All samples were analysed with two approaches, with and without sonication. In total, we obtained 171 C. difficile isolates, which were classified into 25 different ribotypes and five different toxinotypes One of the ribotypes, SLO 302, was newly described. There were no big differences in the number of isolates between the two treatment approaches (sonication, without sonication). But there was a difference in the number of different ribotypes, since with sonication we found 22 different ribotypes, without sonication only nine. The most common ribotypes were 001/072 and 014/020, both of which are toxigenic and belong to toxinotype 0. From the obtained results, we can conclude that the sediments of aquatic environments are commonly contaminated with C. difficile spores and could have potential role in environmental C. difficile transmissions.
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