Potential Role Of Clostridium Difficile And Clostridium Perfringens As A Cause Of Diarrhea In Horses

Farag, Eman F.; Shalaby, Basma; Abd El-Hamed, Taher M.

doi:10.21608/javs.2019.62646

Potential Role Of Clostridium Difficile And Clostridium Perfringens As A Cause Of Diarrhea In Horses

Document Type : Original Article

Authors

¹ Animal Health Research Institute, Bacteriology department, Dokki, Egypt.

² Animal Health Research Institute, Bacteriology department, Dokki, Egypt

³ Faculty of veterinary medicine, Cairo University, Egypt

10.21608/javs.2019.62646

Abstract

Diarrhea is a major clinical problem affecting foals and horses at different ages which can be fatal and cause great economic losses particularly in foals. The aim of this study was to identify Clostridium perfringens and Clostridium difficile involved in enteric infections and the associated virulence factors in diarrheic foals and horses. Thirty fecal samples were examined for identification of C.perfringens and C.difficile (bacteriological culturing, microscopical examination, count and molecular detection). C.perfringens was isolated from young horses less than 6 months age in an incidence of 46.7% with count ranged from 1.3×10⁴- 5.6×10⁸ cfu/g, while from horses of 6 months to one year, it was 20% with count ranged from 1.7×10⁴- 1.9×10⁶ cfu/g, on the other hand, 13.3% with count ranged from 1.4×10³- 2.5×10⁴ cfu/g was the incidence of C.perfringens in horse more than one year. C. difficile was not detected in all examined samples. C.perfringens type A was the most predominant type detected in an incidence of 58.3% then type B (4.7%). All C.perfringens isolates were sensitive to penicillin, nitromedazole, fluramfenicole and ceftiofur and all are resistant to oxytetracycline. Based on the high count, eight C.perfringens isolates were chosen to be molecular characterized for the presence of some virulence factor genes which included ; cpa, cpb2, cpe and netF toxin genes. All examined isolates have cpa (alpha) and cpb2 (beta 2) toxin genes, 5/8 isolates were found to have netF gene while only 2 isolates harbored cpe (enterotoxin gene). C. difficile tpi gene was not detected in all examined samples (10). This study may throw the light on the synergistic effect of β2 and net F toxins together with alpha toxin on induction of enterocolitis in horse.

Keywords

Main Subjects

Bacteriology

References

ANSARI M.R., MOHAMMAD ABADI M.R. AND SHAMSADDINI M. B.2010.Clostridium perfringens isolate typing by multiplex PCR The Journal of Venomous Animals and Toxins including Tropical Diseases 16(4): 573-578

BACCIARINI, L.N, BOERLIN, P., STRAUB, R. 2003. Immuno-histochemical localization of Clostridium perfringens b2-toxin in the gastrointestinal tract of horses. Vet Pathol 40:376–381.

BARR,B.S.,WAIDRIDGE,B.M.,MORRESEY,P.R.,REED,S.M.,CLARK,C.,BEIGRAVE,R.,DONECKER,J.M.AND WEIGET,D.J. 2012. Antimicrobial –associated diarrhea in three equine referral practice. Equine Vetrinary Journal 45 .154-158

DIAB, S.S., SONGER, G., AND UZAL, F.A. 2013A. Clostridium difficile infection in horses, Vet. Microbiol. P (13): 378-1135.

DIAB, S.S., RODRIGUEZ, A., AND UZAL, F.A. 2013 B. Pathology and diagnostic criteria of Clostridium difficile enteric infection in horses.Vet. Pathol.,50(6),1028-1036

EAST, L.M., SAVAGE, C.J., TRAUB-DARGATZ, J.L., DICKINSON, C.E. AND ELLIS, R.P. 1998. Enterocolitis associated with Clostridium perfringens infection in neonatal foals. J. Am. vet. med. Ass. 212, 1751-1756.

FINLEY, A.; MEHDIZADEH GOHARI, I.; PARREIRA, V.R.; ABRAHAMS, M.; STAEMPFLI, H.R. AND PRESCOTT, J.F.2016. Prevalence of netF-positive Clostridium perfringens in foals in southwestern Ontario. Can.J.Vet. Res. 80: 242-244.

FISHER D.J, MIYAMOTO K.,AND HARRISON B. 2005. Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene. Mol. Microbiol 56:747–762.

FREDERICK,J. GIGU`ERE, S. AND SANCHEZ, L. C. 2009. Infectious agents detected in the feces of diarrheic foals: a retrospective study of 233 cases (2003–2008),” Journal of Veterinary Internal Medicine, vol. 23, no. 6, pp. 1254–1260.

FREEDMAN, J.C.; SHRESTHA,A. AND MCCLANE, A.B. 2016. Clostridium perfringens Enterotoxin; Action, Genetics and Translational Applications. Toxins, 8(73): 1-16.

GILBERT,M., JOLIVET-REYNAUD,C.,AND POPOFF, M.R. (1997. Beta 2 toxin ,anovel toxin produced by Clostridium perfringens .Gene 203:65-73

HARRIS, R. SANKAR, K. SMALL, J.-A. SUEPAUL, R. STEWART-JOHNSON, A. AND ADESIYUN, A. 2012. Prevalence and characteristics of enteric pathogens detected in diarrhoeic and non-diarrhoeic foals in trinidad,” Veterinary Medicine International, vol. Article ID 724959, 9 pages

HATHEWAY, C. 1990. Toxigenic clostridia. Clin Microbiol Rev.;3:66-76.

HAZLETT, M. J. KIRCANSKI, J. SLAVIC, D. AND PRESCOTT, J. F. 2011. “Beta 2 toxigenic Clostridium perfringens type A colitis in a three-day old foal,” Journal of Veterinary Diagnostic Investigation, vol. 23, no. 2, pp. 373–376,

HERHOLZ C., MISEREZ R .,AND NICOLET J.1999. Prevalence of b2-toxigenic Clostridium perfringens in horses with intestinal disorders. J. Clin .Microbiol. 37:358–361

KEEL,M.K.,AND SONGER,J.C.(2006):The comparative pathology of Clostridium difficile associated disease.Vet. Pathol.43.225-240

KONEMAN, E.W.; ALLEN,S.D.; DOWELL, V.R. AND SUMMERS, H.W.1992. Colour atlas and text book of diagnostic microbiology. 4^th Ed.J.B.Lippincott, New York,London.

MCDONEL ,J.I. 1986. Toxins of Clostridium perfringens types A,B,C,D and E. Pharmacology of bacterial toxins, In F. Dorner and H. Drews. p. 477- 517, Pergamon press, Oxford, UK.

MEHDIZADEH GOHARI I., ARROYO L., MACINNES J.I., TIMONEY, J.F.,PARREIRA, V.T.,AND PRESCOTT , J.F. 2014. Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis.Can. J. Vet. Res.;78:1–7.

MEHDIZADEH GOHARI ,I., PARREIRA, V.R., NOWELL ,V.J.,NICHOLSON, V.M., OLIPHANT, K., AND PRESCOTT, J.F. 2015. 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 10(4):e0122684. doi:10.1371/journal.pone.0122684

MEHDIZADEH GOHARI, I., KROPINSKI ,A.M., AND WEESE, S.J.2016. Plasmid characterization and chromosome analysis of two netF+ Clostridium perfringens isolates associated with foal and canine necrotizing enteritis. PLoS One.;11(2):e0148344.

NATIONAL COMMITTEE FOR CLINICAL LABORATORY STANDARDS (NCCL)2002. GINOCCHIOCC. AM.J.HEALTH SYST.PHARM.

RAJABALLY, N.; KULLIN, B.; EBRAHIM, K.; BROCK, T.; WEINTRAUB, A.; WHITELAW, A.; BAMFORD, C.; WATERMEYER, G.; THOMSON, S.; ABRATT, V. AND REID, S. 2016. A comparison of Clostridium difficile diagnostic methods for identification of local strains in a South African centre.

RADOSTITS, O.M. GAY, C. C. HINCHCLIFF, K.W. AND CONSTABLE, P. D.2007. Acute diarrhea in suckling foals,” in Veterinary medicine Publishing Group-A Textbook of the diseases of Cattle, Horses, Sheep, Pigs, and Goats, OM. Radostits, C. C. Gay, K.W. Hinchcliff, and P. D. Constable, Eds., pp. 274–277, Saunders, Philadelphia, Pa,

USA, 10th edition.

QUINN, P. J. MARKEY, B. K. DONNELLY, W. J. LEONARD, F. C. FANNING, S.AND MAGUIRE, D.2011. Veterinary Microbiology and MicrobialDisease, Blackwell Publishing House Science LTDA, Oxford,UK.

SCHOSTER,A.;GUILLERMO,A.L.;STAEMPFLI,H.R.;SHEWEN,P.E.AND WEESE,J.S.2012. Presence and molecular characterization of Clostridium difficile and Clostridium perfringens in intestinal compartments of healthy horses.BMC Vetrinary Research ;8:94

SILVA, R.O.S.; RIBEIRO, M.G. AND PALHARES, M.S.2013. Detection of A/B toxin and isolation of Clostridium difficle and Clostridium perfringens from foals. Equine Vet. J. (45) 6:671-675.

SLOVIS, N. M. ELAM, J. ESTRADA, M. THAO, . M. F. AND LEUTENEGGER, C. M.2010. Comprehensive analysis of infectious agents associated with diarrhea in foals in central Kentucky, in Proceedings of the Annual Convention of the AAEP, Baltimore, Md, USA,

SONGER JG. (1996): Clostridial enteric diseases of domestic animals. Clin Microbiol Rev;9:216–234. 22

STERN ,M.AND BATTY,I.(1975): Pathogenic Clostridia Butter Worth, London Boston

THEAN, S., ELLIOTT, B.,AND RILEY,T.V. 2011. Clostridium difficile in horses in Australia--a preliminary study. Division of Microbiology and Infectious Diseases, J Med Microbiol. Aug; 60 (Pt 8):1188-92

UZAL,F.A .2004. Diagnosis of C.perfringens intestinal infection in sheep and goat .Anaerobe V(10) 2: 135-143

WAGGETT,B.E.,MCGORUM ,B.C. ,WERNERY ,V ,SHAW ,D.J. AND PIRIE ,R.S. 2010. prevalence of Clostridium perfringens in faces and ileal contents from grass sickness affected horses: comparisons with three control populations Equine Vet.J.,42 (6) :494-499

WEESE, J.S., STAEMPFLI, H.R., ANDPRESCOTT, J.F.( 2001): A prospective study of the roles of Clostridium difficile and enterotoxigenic Clostridium perfringens in equine diarrhea. Equine ,Vet. J. 33:403–409.

VAN ASTEN, A.J.; VAN DER WIEL, C.W.; NIKOLAOU, G.; HOUWERS, D.J. AND GRÖNE, A.2009. A multiplex PCR for toxin typing of Clostridium perfringens isolates. Vet Microbiol. 136(3-4):411-2.

VEDANTAM,C.,CLARK,A.,CHU,M.,MCQUADE,R.,MALLOZZI,M.,AND VISWANATHAN,V.K. 2012. Clostridium difficile infection :Toxins and non –toxin ,virulence factors and their contributions to disease establishment and most response, Gut Microbes 3,121-134

VILEI, E.M., SCHLATTER, Y., PERRETEN, V., STRAUB, R., POPOFF ,M.R., AND GIBERT, M.2005. Antibiotic-induced expression of a cryptic cpb2 gene in equine beta2-toxigenic Clostridium perfringens. Mol Microbiol 57: 1570–1581. PMID: 16135225