Genotypic and Phenotypic Characterization ofClostridium perfringensandClostridium difficilein Diarrheic and Healthy Dogs

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Abstract
The objectives of this study were to examine the potential roles ofClostridium difficileand enterotoxigenicClostridium perfringensin diarrhea in dogs by comparison of isolation, determination of toxin status via enzyme‐linked immunosorbent assay (ELISA), and application of multiplex polymerase chain reaction (PCR). These techniques were used to evaluate fecal specimens in 132 healthy and diarrheic dogs. These dogs were prospectively evaluated by grouping them into the following 3 categories: hospitalized dogs with diarrhea (n = 32), hospitalized dogs without diarrhea (n = 42), and apparently healthy outpatient dogs without diarrhea (n = 58). All fecal specimens were cultured using selective media forC difficile, Salmonellaspp., andCampylobacterspp. and selective media after heat shock forC perfringens.No significant difference was found in the isolation ofC perfringensorC difficileamong the 3 groups. A significant association was found between the presence of diarrhea and detection ofC perfringensenterotoxin (CPE) or toxin A via ELISA for bothC perfringensandC difficile, respectively. PCR performed onC difficileisolates for toxin A and toxin B genes revealed no significant differences among the 3 groups, but diarrheic dogs were significantly more likely to be positive for the enterotoxin gene ofC perfringens.Based on the results of this study, the use of ELISA for detection of CPE in feces combined with the detection of enterotoxigenic fecal isolates obtained via heat shock provides the strongest evidence for the presence ofC perfrigences‐associated diarrhea.