FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Can J Vet Res / Characterization of Clostridium perfringens in the feces of ...
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).
Get Full Text
Publication Date: 2014-01-08 PubMed ID: 24396174PubMed Central: PMC3878003
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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

References

This article includes 22 references
  1. Feary DJ, Hassel DM. Enteritis and colitis in horses.. Vet Clin North Am Equine Pract 2006;22:437–479.
    pubmed: 16882483
  2. Rooney JR, Bryans JT, Doll ER. Colitis “X” of horses.. J Am Vet Med Assoc 1963;142:510–511.
    pubmed: 13974694
  3. Staempfli HR, Townsend HGG, Prescott JF. Prognostic features and clinical presentation of acute idiopathic enterocolitis in horses.. Can J Vet Res 1991;32:232–237.
    pmc: PMC1481315pubmed: 17423769
  4. Båverud V. Clostridium difficile in horses.. PhD dissertation. Uppsala, Sweden: Swedish University of Agricultural Sciences; 2002.
  5. Weese JS, Toxopeus L, Arroyo L. Clostridium difficile associated diarrhea in horses within the community: Predictors, clinical presentation and outcome.. Equine Vet J 2006;38:185–188.
    pubmed: 16536390
  6. Ruby R, Magdesian KG, Kass PH. Comparison of clinical, microbiologic, and clinicopathologic findings in horses positive and negative for Clostridium difficile infection.. J Am Vet Med Assoc 2009;234:777–784.
    pubmed: 19284345
  7. Kircanski J, Hodgins D, Soltes G. Development of an antigen-capture enzyme-linked immunosorbent assay for Clostridium perfringens beta2-toxin in porcine feces and the neonatal piglet intestine.. J Vet Diagn Invest 2012;24:895–902.
    pubmed: 22855377
  8. Farzan A, Kircanski J, DeLay J. An investigation into the association between cpb2-encoding Clostridum perfringens type A and diarrhea in neonatal piglets.. Can J Vet Res 2013;77:45–53.
    pmc: PMC3525171pubmed: 23814355
  9. Tillotson K, Traub-Dargatz JL, Dickinson CE. Population-based study of fecal shedding of Clostridium perfringens in broodmares and foals.. J Am Vet Med Assoc 2002;220:342–348.
    pubmed: 11829266
  10. Donaldson MT, Palmer JE. Prevalence of Clostridium perfringens enterotoxin and Clostridium difficile toxin A in feces of horses with diarrhea and colic.. J Am Vet Med Assoc 1999;215:358–361.
    pubmed: 10434974
  11. Frederick J, Giguère S, Sanchez LC. Infectious agents detected in the feces of diarrheic foals: A retrospective study of 233 cases (2003–2008). J Vet Intern Med 2009;23:1254–1260.
    pmc: PMC7166729pubmed: 19747192
  12. Weese JS, Staempfli HR, Prescott JF. A prospective study of the roles of Clostridium difficile and enterotoxigenic Clostridium perfringens in equine diarrhea.. Equine Vet J 2001;33:403–409.
    pubmed: 11469775
  13. Van Baelen D, Devriese LA. Presence of Clostridium perfringens enterotoxin in intestinal samples from farm animals with diarrhea of unknown origin.. Zentralbl Veterinarmed B 1987;34:713–716.
    pubmed: 2894102
  14. Kanoe M, Inoue S, Abe T. Isolation of Clostridium perfringens from foals.. Microbios 1990;64:153–158.
    pubmed: 2084493
  15. Herholz C, Miserez R, Nicolet J. Prevalence of beta2- toxigenic Clostridium perfringens in horses with intestinal disorders.. J Clin Microbiol 1999;37:358–361.
    pmc: PMC84307pubmed: 9889218
  16. Bacciarini LN, Boerlin P, Straub R, Frey J, Grone A. Immunohistochemical localization of Clostridium perfringens beta2-toxin in the gastrointestinal tract of horses.. Vet Pathol 2003;40:376–381.
    pubmed: 12824509
  17. Jost BH, Billington SJ, Trinh HT, Bueschel DM, Songer JG. Atypical cpb2 genes, encoding beta2-toxin in Clostridium perfringens isolates of non-porcine origin.. Infect Immun 2005;73:652–656.
    pmc: PMC538998pubmed: 15618211
  18. Fisher DJ, Miyamoto K, Harrison B, Akimoto S, Sarker MR, McClane BA. Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene.. Mol Microbiol 2005;56:747–762.
    pubmed: 15819629
  19. Diab SS, Kinde H, Moore J. Pathology of Clostridium perfringens Type C enterotoxemia in horses.. Vet Pathol 2012;49:255–263.
    pubmed: 21502373
  20. Traub-Dargatz JL, Jones RL. Clostridia-associated enterocolitis in adult horses and foals.. Vet Clin North Am Equine Pract 1993;9:411–421.
    pubmed: 8358653
  21. Songer JG. Clostridial enteric diseases of domestic animals.. Clin Microbiol Rev 1996;9:216–234.
    pmc: PMC172891pubmed: 8964036
  22. McGorum BC, Dixon PM, Smith DGE. Use of metronidazole in equine acute idiopathic toxaemic colitis.. Vet Rec 1998;142:635–638.
    pubmed: 9650235

Citations

This article has been cited 18 times.
  1. Hardefeldt L, Thomas K, Page S, Norris J, Browning G, El Hage C, Stewart A, Gilkerson J, Muscatello G, Verwilghen D, van Galen G, Bauquier J, Cuming R, Reynolds B, Whittaker C, Wilkes E, Clulow J, Burden C, Begg L. Antimicrobial prescribing guidelines for horses in Australia. Aust Vet J 2025 Dec;103(12):781-889.
    doi: 10.1111/avj.70003pubmed: 40903020google scholar: lookup
  2. Haywood LMB, Sheahan BJ. A Review of Epithelial Ion Transporters and Their Roles in Equine Infectious Colitis. Vet Sci 2024 Oct 7;11(10).
    doi: 10.3390/vetsci11100480pubmed: 39453072google scholar: lookup
  3. Tang Z, Li X, Wang X, Zhang C, Zou L, Ren H, Liu W. Characterization and Genomic Analysis of a Novel Lytic Phage DCp1 against Clostridium perfringens Biofilms. Int J Mol Sci 2023 Feb 20;24(4).
    doi: 10.3390/ijms24044191pubmed: 36835606google scholar: lookup
  4. Serroni A, Colabella C, Cruciani D, Ciullo M, Crotti S, Papa P, Di Paolo A, Gobbi M, Forti K, Pellegrini M, Salini R, D'Avino N, Cagiola M, Pezzotti G, De Giuseppe A. Identification and Characterization of Clostridium perfringens Atypical CPB2 Toxin in Cell Cultures and Field Samples Using Monoclonal Antibodies. Toxins (Basel) 2022 Nov 17;14(11).
    doi: 10.3390/toxins14110796pubmed: 36422970google scholar: lookup
  5. Gomez DE, Leclere M, Arroyo LG, Li L, John E, Afonso T, Payette F, Darby S. Acute diarrhea in horses: A multicenter Canadian retrospective study (2015 to 2019). Can Vet J 2022 Oct;63(10):1033-1042.
    pubmed: 36185796
  6. Benz R, Piselli C, Hoxha C, Koy C, Glocker MO, Popoff MR. Clostridium perfringens Beta2 toxin forms highly cation-selective channels in lipid bilayers. Eur Biophys J 2022 Jan;51(1):15-27.
    doi: 10.1007/s00249-021-01577-7pubmed: 34854958google scholar: lookup
  7. Willette JA, Kopper JJ, Kogan CJ, Seguin MA, Schott HC. Effect of season and geographic location in the United States on detection of potential enteric pathogens or toxin genes in horses ≥6-mo-old. J Vet Diagn Invest 2022 May;34(3):407-411.
    doi: 10.1177/10406387211056054pubmed: 34763559google scholar: lookup
  8. Kopper JJ, Willette JA, Kogan CJ, Seguin A, Bolin SR, Schott HC 2nd. Detection of pathogens in blood or feces of adult horses with enteric disease and association with outcome of colitis. J Vet Intern Med 2021 Sep;35(5):2465-2472.
    doi: 10.1111/jvim.16238pubmed: 34382708google scholar: lookup
  9. Owens LA, Colitti B, Hirji I, Pizarro A, Jaffe JE, Moittié S, Bishop-Lilly KA, Estrella LA, Voegtly LJ, Kuhn JH, Suen G, Deblois CL, Dunn CD, Juan-Sallés C, Goldberg TL. A Sarcina bacterium linked to lethal disease in sanctuary chimpanzees in Sierra Leone. Nat Commun 2021 Feb 3;12(1):763.
    doi: 10.1038/s41467-021-21012-xpubmed: 33536429google scholar: lookup
  10. Wenzel CQ, Mills DC, Dobruchowska JM, Vlach J, Nothaft H, Nation P, Azadi P, Melville SB, Carlson RW, Feldman MF, Szymanski CM. An atypical lipoteichoic acid from Clostridium perfringens elicits a broadly cross-reactive and protective immune response. J Biol Chem 2020 Jul 10;295(28):9513-9530.
    doi: 10.1074/jbc.RA119.009978pubmed: 32424044google scholar: lookup
  11. Taylor S. A review of equine sepsis. Equine Vet Educ 2015 Feb;27(2):99-109.
    doi: 10.1111/eve.12290pubmed: 32313390google scholar: lookup
  12. Mehdizadeh Gohari I, Unterer S, Whitehead AE, Prescott JF. NetF-producing Clostridium perfringens and its associated diseases in dogs and foals. J Vet Diagn Invest 2020 Mar;32(2):230-238.
    doi: 10.1177/1040638720904714pubmed: 32081091google scholar: lookup
  13. Park CS, Hwang JY, Cho GJ. The First Identification and Antibiogram of Clostridium perfringens Type C Isolated from Soil and The Feces of Dead Foals in South Korea. Animals (Basel) 2019 Aug 20;9(8).
    doi: 10.3390/ani9080579pubmed: 31434197google scholar: lookup
  14. Kiu R, Hall LJ. An update on the human and animal enteric pathogen Clostridium perfringens. Emerg Microbes Infect 2018 Aug 6;7(1):141.
    doi: 10.1038/s41426-018-0144-8pubmed: 30082713google scholar: lookup
  15. Shaw SD, Stämpfli H. Diagnosis and Treatment of Undifferentiated and Infectious Acute Diarrhea in the Adult Horse. Vet Clin North Am Equine Pract 2018 Apr;34(1):39-53.
    doi: 10.1016/j.cveq.2017.11.002pubmed: 29426709google scholar: lookup
  16. Olivo G, Lucas TM, Borges AS, Silva RO, Lobato FC, Siqueira AK, da Silva Leite D, Brandão PE, Gregori F, de Oliveira-Filho JP, Takai S, Ribeiro MG. Enteric Pathogens and Coinfections in Foals with and without Diarrhea. Biomed Res Int 2016;2016:1512690.
    doi: 10.1155/2016/1512690pubmed: 28116290google scholar: lookup
  17. Finley A, Gohari IM, Parreira VR, Abrahams M, Staempfli HR, Prescott JF. Prevalence of netF-positive Clostridium perfringens in foals in southwestern Ontario. Can J Vet Res 2016 Jul;80(3):242-4.
    pubmed: 27408339
  18. Mehdizadeh Gohari I, Parreira VR, Nowell VJ, Nicholson VM, Oliphant K, Prescott JF. A novel pore-forming toxin in type A Clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis. PLoS One 2015;10(4):e0122684.
    doi: 10.1371/journal.pone.0122684pubmed: 25853427google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.