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Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2014; 78(1); 1-7;

Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis.

Abstract: Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered. Jusqu’à 60 % des cas de colite équine n’ont aucune cause connue. Afin d’améliorer la compréhension des causes de colite aigüe chez les chevaux, nous émettons l’hypothèse que les Clostridium perfringens produisant l’entérotoxine (CPE) et/ou la toxine bêta-2 (CPB2) sont des causes courantes et importantes de colites sévères chez les chevaux et/ou qu’une cytotoxine non décrite encore produite par C. perfringens pourrait également causer des colites chez les chevaux. Des échantillons fécaux provenant de 55 chevaux (43 adultes, 12 poulains) avec des évidences cliniques de colite ont été évalués par culture pour la présence de Clostridium difficile, C. perfringens et Salmonella. Les fèces furent également analysées par épreuve immuno-enzymatique (ELISA) pour la présence des toxines A/B de C. difficile et les toxines alpha (CPA), bêta2 (CPB2) et l’entérotoxine (CPE) de C. perfringens. Cinq isolats de C. perfringens par échantillons ont été typés pour les gènes suivants : cpa, cpb, cpb2 consensus, cpb2 atypique, cpe (entérotoxine), etx (toxine epsilon), itx (toxine iota), netB (toxine B de l’entérite nécrotique), et tpeL (cytotoxine large de C. perfringens). Les surnageants de culture de ces isolats ont également été évalués pour leur cytotoxicité envers une lignée cellulaire équine. Tous les échantillons fécaux étaient négatifs pour la présence de Salmonella. Clostridium perfringens et C. difficile furent isolés de 40 % et 5,4 % des échantillons, respectivement. Tous les échantillons de fèces étaient négatifs pour CPE. Les toxines CPA et CPB2 furent détectées à partir de 14,5 % et 7,2 % des échantillons fécaux, respectivement, tous étant positifs pour la présence de C. perfringens en culture. Aucun des isolats n’était positif pour la présence des gènes cpe, etx, netB ou tpeL. Les gènes cpb2 atypiques et cpb2 consensus furent identifiés dans respectivement 15 (13,6 %) et 4 (3,6 %) des 110 isolats. Tous les isolats de C. perfringens équins montraient des effets cytotoxiques nettement plus légers que les témoins positifs produisant CPB, bien que les isolats possédant les gènes cpb2 étaient légèrement mais significativement plus cytotoxiques que les témoins négatifs. En fonction de la population étudiée, il nous est impossible de confirmer notre hypothèse qu’en Ontario des C. perfringens produisant CPE et CPB-2 sont courants chez les chevaux avec colites et nous n’avons pas réussi à identifier une activité cytotoxique in vitro chez les types A isolés.(Traduit par Docteur Serge Messier).
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Publication Date: 2014-01-08 PubMed ID: 24396174PubMed Central: PMC3878003
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study investigates the proposed hypothesis that bacteria Clostridium perfringens, which produces certain toxins, may be a common cause of severe colitis in horses. However, the results of the study did not confirm the hypothesis, failing to find significant presence or toxicity of the said bacteria in fecal samples from horses with colitis.

Objective and Hypothesis

  • The study sets out to address the fact that up to 60% of equine colitis cases have no known cause.
  • The researchers propose a hypothesis that bacteria named Clostridium perfringens, particularly types that produce enterotoxin and beta2 toxin, might be commonly responsible for severe colitis in horses.
  • They also propose that a yet unidentified cytotoxin produced by C. perfringens could also be a cause for colitis in horses.

Methodology

  • The study involved the analysis of fecal samples from 55 horses (including 43 adults and 12 foals) exhibiting clinical signs of colitis.
  • The samples were evaluated for the presence of certain bacteria (Clostridium difficile, C. perfringens, and Salmonella) using culture methods.
  • Enzyme-linked immunosorbent assay (ELISA) tests were used to examine the samples for C. difficile A/B toxins and three C. perfringens toxins (alpha toxin, beta2 toxin, and enterotoxin).
  • Moreover, five C. perfringens isolates from each sample were genotyped to further examine various toxin-producing genes.
  • The isolated samples were also tested for their toxicity on an equine cell line.

Study Findings

  • None of the fecal samples tested positive for Salmonella.
  • C. perfringens was found in 40% of the samples, whereas C. difficile was found in just 5.4% samples.
  • None of the samples contained C. perfringens enterotoxin.
  • C. perfringens alpha toxin and beta2 toxin were found in 14.5% and 7.2% of samples, respectively. These were all culture-positive for C. perfringens.
  • No toxin-producing genes (cpe, etx, netB, or tpeL) were identified in the isolates.
  • Certain beta2 genes were identified in a small proportion of the isolates, but all equine C. perfringens isolates exhibited milder cytotoxic effects compared to a CPB-producing control. Isolates with such beta2 genes were somewhat more cytotoxic than negative isolates.

Conclusion

  • Given the results, the study was unable to endorse its initial hypothesis that CPE and CPB2-producing C. perfringens bacteria are common causes of colitis in horses, at least in Ontario.
  • The research did not identify any substantial cytotoxic activity in the type A C. perfringens isolates received.

Cite This Article

APA
Gohari IM, Arroyo L, Macinnes JI, Timoney JF, Parreira VR, Prescott JF. (2014). Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis. Can J Vet Res, 78(1), 1-7.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 1928-9022
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 78
Issue: 1
Pages: 1-7

Researcher Affiliations

Gohari, Iman Mehdizadeh
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).
Arroyo, Luis
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).
Macinnes, Janet I
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).
Timoney, John F
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).
Parreira, Valeria R
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).
Prescott, John F
  • Department of Pathobiology (Gohari, MacInnes, Parreira, Prescott) and Department of Clinical Studies (Arroyo), University of Guelph, Guelph, Ontario N1G 2W1; Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA (Timoney).

MeSH Terms

  • Acute Disease
  • Aging
  • Animals
  • Clostridium Infections / microbiology
  • Clostridium Infections / veterinary
  • Clostridium perfringens / classification
  • Clostridium perfringens / isolation & purification
  • Enterocolitis / microbiology
  • Enterocolitis / veterinary
  • Feces / microbiology
  • Horse Diseases / microbiology
  • Horses

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