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Clostridium difficile is a Gram-positive and sporulating enteropathogen that is a major cause of healthcare-associated infections. Even though a large number of genomes of this species have been sequenced, only a few plasmids have been described in the literature. Here, we use a combination of in silico analyses and laboratory experiments to show that plasmids are common in C. difficile. We focus...
Recurrent Clostridium difficile infection (CDI) is an increasing problem, yet reasons for this are poorly understood. Attention has been paid to the role of strain, with conflicting association of ribotype 027 and recurrences.Stool samples and medical records data were collected from 60 patients: 27 with recurrent CDI and 33 with single episode CDI. C. difficile was isolated and ribotyped, and minimum...
Commercial nucleic acid extraction kits are a cost effective, efficient and convenient way to isolate DNA and RNA from bacteria. Despite the increasing importance of the gastrointestinal pathogen, Clostridium difficile, and the increased use of nucleic acids in its identification, characterization, and investigation of virulence factors, no standardized or recommended methods for nucleic acid isolation...
Clostridium difficile is the primary known cause of antibiotic-associated diarrhea. Diarrheal disease in food animals due to C. difficile infection has been well documented. Recently, reports of C. difficile infections in patients with no known risk factors for disease have raised concern of community acquisition through food animals and food. In this study, multi-locus variable number tandem repeat...
The objective of this study was to determine the prevalence of Clostridium difficile contamination in retail seafood and fish from Canadian grocery stores. C. difficile was found in 4.8% (5/119) of the samples. This study, combined with studies of other food sources, suggests that widespread contamination of food is common.
A longitudinal study of Clostridium difficile colonization in piglets was performed on a conventional swine farm in Ontario, Canada. Fecal samples were collected from 10 sows prior to their expected farrowing date, and then from all their piglets on days 2, 7, 30, 44 and 62 of life. C. difficile was isolated from 4/10 (40%) of sows prior to farrowing, 90/121 (74%) piglets on day 2, 66/117 (56%) on...
We quantified the thermal inhibitory effect of 71°C (recommended for cooking ground meats), and re-heating at 85°C, on food- and food-animal-derived Clostridium difficile spores. All C. difficile strains tested (n=20) survived 71°C for 2h, but 90% died within 10min when re-heated at 85°C. Current cooking recommendations would need revision to include C. difficile.
Quantitative real-time polymerase chain reaction (qPCR) is a sensitive, efficient and reproducible technique for studying gene expression. Identification of stably expressed reference genes is required to avoid bias in these studies yet mostly unvalidated reference genes are used in studying gene expression in Clostridium difficile. Here, we sought to identify a set of stable reference genes used...
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