High contamination rates of shoes of veterinarians, veterinary support staff and veterinary students with Clostridioides difficile spores.

Clostridioides difficile is often found in animals and their environment. However, not much has been reported on veterinary clinics environment in terms of the spore load, prevalence and PCR ribotype diversity. The aim of this study was to assess the prevalence of C. difficile on shoe soles of veterinarians, veterinary support staff and veterinary students at the Veterinary Faculty campus. Altogether, 50 shoe sole swabs were collected, and the positivity rates ranged from 86.7% in swabs from veterinarians to 100% in swabs from support staff and students. Non-toxigenic and toxigenic strains representing toxinotypes 0, IV and XIX were isolated and distributed into 17 different PCR ribotypes, most common being 010, 014/020, SLO002 and 009. PCR ribotype 010 was the most prevalent and isolated from shoe soles sampled in 6/7 areas. Students' shoes had highest ribotype diversity (15/17 PCR ribotypes) but showed a low overlap with ribotype isolated from vets and support staff shoes. Veterinary students are likely the main vectors of C. difficile spores transmissions among veterinary teaching clinics and the hospital.

MRSA diversity and the emergence of LA-MRSA in a large teaching hospital in Slovenia.

The methicillin-resistant Staphylococcus aureus (MRSA) is one of the major causes of a variety of infections in hospitals and the community. One of the most prominent changes in the MRSA epidemiology is the emergence of livestock-associated MRSA (LA-MRSA) strains in the human population. The aim of this study was to follow the MRSA epidemiology in a large teaching hospital during an 8-year time period (2006-2013). Altogether 519 MRSA, cultured from screening or clinical samples, were distributed into 77 spa types, of which three (t003 and t001, associated with CC5; and t015; associated with CC45) were the most common. LA-MRSA-associated spa types (t011, t034, t108, t899; associated with CC398) started to emerge in the year 2009 and continued to be found annually at a frequency from 3.9% to 12.7% of all MRSA strains examined. Only 6 of 27 LA-MRSA strains were associated with infections.

Distribution of Clostridium difficile PCR ribotypes and high proportion of 027 and 176 in some hospitals in four South Eastern European countries.

While Clostridium difficile epidemiology is well documented in many European countries, data are largely missing for South Eastern European region. Here we report the PCR ribotype distribution of 249 C. difficile isolates received for typing from six hospital settings from Croatia, Bosnia and Herzegovina, Republic of Macedonia and Serbia in time period from 2008 to 2015. Twenty-four PCR ribotypes were detected. The majority of strains from Bosnia and Herzegovina and Serbia belonged to PCR ribotype 027 (65.8%). Other three dominating PCR ribotypes were 176 (18 strains; Croatia), 001/072 (15 strains; all countries) and 014/020 (15 strains; all countries).

Clostridium difficile and Clostridium perfringens from wild carnivore species in Brazil.

Despite some case reports, the importance of Clostridium perfringens and Clostridium difficile for wild carnivores remains unclear. Thus, the objective of this study was to identify C. perfringens and C. difficile strains in stool samples from wild carnivore species in Brazil. A total of 34 stool samples were collected and subjected to C. perfringens and C. difficile isolation. Suggestive colonies of C. perfringens were then analyzed for genes encoding the major C. perfringens toxins (alpha, beta, epsilon and iota) and the beta-2 toxin (cpb2), enterotoxin (cpe) and NetB (netb) genes. C. difficile strains were analyzed by multiplex-PCR for toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB) and also submitted to a PCR ribotyping. Unthawed aliquots of samples positive for C. difficile isolation were subjected to the detection of A/B toxins by a cytotoxicity assay (CTA). C. perfringens was isolated from 26 samples (76.5%), all of which were genotyped as type A. The netb gene was not detected, whereas the cpb2 and cpe genes were found in nine and three C. perfringens strains, respectively. C. difficile was isolated from two (5.9%) samples. A non-toxigenic strain was recovered from a non-diarrheic maned wolf (Chrysocyon brachyurus). Conversely, a toxigenic strain was found in the sample of a diarrheic ocelot (Leopardus pardallis); an unthawed stool sample was also positive for A/B toxins by CTA, indicating a diagnosis of C. difficile-associated diarrhea in this animal. The present work suggests that wild carnivore species could carry C. difficile strains and that they could be susceptible to C. difficile infection.

High diversity of Clostridium difficile genotypes isolated from a single poultry farm producing replacement laying hens.

Clostridium difficile is well established as a pathogen of horses, calves, and pigs, but little is known about its prevalence in poultry. In this study, chicken fecal samples were collected on four occasions from two populations being raised as layer replacements. Samples were examined by an enrichment culture method, and 38 of 61 (62.3%) were culture positive. The rate of colonization seemed to be age dependent: 100% of fecal samples from 2-week- old birds were culture positive, and the colonization rate decreased to 71.4% in 14-week- old birds, and to 40.9% in 18-week- old birds. Unlike in other domestic animal hosts, the diversity of PCR ribotypes found on a single farm was high, and 44 isolated strains belonged to 12 PCR ribotypes. Furthermore, the prevalence of variant strains having changed toxin genes for toxins A and B and possessing an additional toxin, binary toxin, was low. The majority of strains were toxinotype 0, although two were nontoxinogenic and three were binary toxin-positive toxinotype IV. Toxinotype V strains, which are often associated with food animals, were not found.