| Summary: | The role of <i>Clostridioides (C.) difficile</i> as an enteropathogen in dogs is controversial. In humans, intestinal bile acid-dysmetabolism is associated with <i>C. difficile</i> prevalence. The relationship between fecal qPCR-based dysbiosis index (DI) and especially the abundance of bile acid-converting <i>Clostridium hiranonis</i> with the presence of <i>C. difficile</i> in dogs was explored across the following 4 cohorts: 358 fecal samples submitted for routine diagnostic work-up, 33 dogs with chronic enteropathy, 14 dogs with acute diarrhea, and 116 healthy dogs. Dogs that tested positive for <i>C. difficile</i> had significantly higher DI (median, 4.4 (range from 0.4 to 8.6)) and lower <i>C. hiranonis</i> (median, 0.1 (range from 0.0 to 7.5) logDNA/g) than dogs that tested negative for <i>C. difficile</i> (median DI, −1 (range from −7.2 to 8.9); median <i>C. hiranonis</i> abundance, 6.2 (range from 0.1 to 7.5) logDNA/g; <i>p</i> < 0.0001, respectively). In 33 dogs with CE and 14 dogs with acute diarrhea, the treatment response did not differ between <i>C. difficile</i>-positive and -negative dogs. In the group of clinically healthy dogs, 9/116 tested positive for <i>C. difficile</i>, and 6/9 of these had also an abnormal DI. In conclusion, <i>C. difficile</i> is strongly linked to intestinal dysbiosis and lower <i>C. hiranonis</i> levels in dogs, but its presence does not necessitate targeted treatment.
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