Isolation and genotyping of Clostridium perfringens and Clostridium difficile in Capuchin Monkeys (Sapajus spp.)

Abstract

The importance of Clostridium perfringens and C. difficile for most wild animal species remains unclear. This study aimed to isolate and genotype C. perfringens and C. difficile in stool samples from free-living and captive capuchin monkeys (Sapajus flavius and Sapajus libidinosus) in Brazil. Ten free-living S. flavius and 14 captive S. libidinosus were sampled for this study. To isolate C. difficile, stool samples were inoculated on plates containing cycloserine-cefoxitin fructose agar supplemented with horse blood and sodium taurocholate. Two different protocols for C. perfringens isolation were tested: direct plating onto selective agar and enrichment in brain heart infusion (BHI) broth followed by plating onto selective agar. C. difficile was not detected in the present study. The results were identical for both protocols tested for isolation of C. perfringens. Four samples (16.7%) were positive for C. perfringens type A, including one sample from a free-living animal (4.2%) and three from captive animals (12.5%), meaning there was no significant difference between these two groups. C. perfringens isolates were negative for all additional virulence factors evaluated, including enterotoxin encoding-gene (cpe) and beta-2 encoding-gene (cpb2). These results suggested that C. perfringens type A is found in the microbiota of capuchin monkeys, although it is less frequent than previously reported in domestic animals.

Occurrence of Clostridium perfringens in Wild Mammals in the Amazon Biome

“…The finding that more than 60% of the samples were positive for C. perfringens type A corroborates the findings of previous studies of other species of wild animals, showing that type A is the most prevalent commensal [9,15,[17][18][19][20]. Some species included in the present study were negative for this anaerobic microorganism.…”

“…Most previous studies that investigated the prevalence of C. perfringens in wild animals have focused on a single animal species or on groups belonging to the same family and with similar feeding habits [9,15,[17][18][19]]. This appears to be the first study covering this diversity of forest mammals within Brazil, allowing a better understanding of the occurrence of C. perfringens in the Amazon biome.…”

“…In humans, it is the second 2 most common cause of foodborne illness in the United States and the sixth most common in Brazil [5]. Some studies have demonstrated the presence of C. perfringens as a commensal in some species of wild animal in addition to confirming the participation of this agent as a cause of enteritis and enterotoxaemia [6][7][8][9][10]. However, the description of this bacterium and its toxinotypes in wild animals of the Amazon Biome, either as a commensal or as a pathogen, is lacking.…”