Utvrđivanje prisutnosti enterotoksina bakterije clostridium perfringens u fecesu zdravih pasa i pasa s proljevom

master's thesis

• Title (english): Presence of Clostridium Perfringens Enterotoxin in the Feces of Healthy Dogs and Dogs with Diarrhea
• Author: Stella Lukman
• Mentor: Suzana Hađina (mentor)
• Committee member: Ljubo Barbić (predsjednik povjerenstva)
• Committee member: Vladimir Stevanović (član povjerenstva)
• Committee member: Suzana Hađina (član povjerenstva)
• Granter: University of Zagreb; Faculty of Veterinary Medicine; (Department of Microbiology and Infectious Diseases with Clinic); Zagreb
• Defense date and country: 2019-04-05, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Veterinary Medicine; Veterinary Clinical Sciences

Abstract

Enterotoksin bakterije Clostridium perfringens često je povezan s akutnim i kroničnim proljevima te sindromom akutnog hemoragičnog proljeva u pasa, no nastanak proljeva uzrokovanog njegovim enterotoksinom do danas nije razjašnjen. Ne postoji zlatni standard za potvrđivanje prisutnosti bakterije C. perfringens u fecesu životinja s proljevom, a dijagnoza se postavlja na temelju kliničke slike, nalaza veće količine C. perfringens endospora u razmazu fecesa ili utvrđivanjem enterotoksina korištenjem imunoenzimnog testa (ELISA, eng. enzyme linked immunosorbent assay) zajedno s molekularnom metodom lančane reakcije polimerazom (PCR, eng. polymerase chain reaction) iz uzoraka fecesa. Cilj ovog istraživanja bio je ustanoviti prisutnost enterotoksina bakterije C. perfringens u fecesu zdravih pasa i pasa s proljevom korištenjem ELISA testa i PCR metodom. Uzorci fecesa sakupljeni su od sveukupno 82 pasa, 40 zdravih i 42 s proljevom nepoznate etiologije. ELISA testom utvrđena je prisutnost C. perfringens enterotoksina (CPE, eng. Clostridium perfringens enterotoxin) u 40% (17/42) pasa s proljevom i 12,5% (5/40), zdravih pasa. Prisutnost CPE nije bila statistički značajno veća kod pasa s proljevom (p=0,3179) u odnosu na zdrave pse. Prevalencija cpe gena utvrđenog PCR metodom bila je 50% (21/42) kod pasa s proljevom i 70% (28/40) kod zdravih pasa. Prisutnost cpe gena nije bila statistički značajno viša kod pasa s proljevom u odnosu na zdrave pse (p=0,0903). Uzorci fecesa pasa s proljevom bili su pozitivni na CPE i na cpe gen u 26% slučajeva (11/42), te u 10% (4/40) zdravih pasa. Statističkom obradom uočena je značajna razlika između rezultata dobivenih PCR metodom naspram onih dobivenih ELISA testom (p=0,0072). U provedenom istraživanju enterotoksin bakterije C. perfringens utvrđen je osim u pasa s proljevom i u zdravih pasa. Rezultati ovog istraživanja pokazali su da je za pouzdanu dijagnostiku enterotoksina bakterije Clostridium perfringens potrebno koristiti kombinaciju PCR i ELISA metode.

Abstract (english)

Clostridium perfringens enterotoxin is often associated with acute and chronic diarrhea as well as with acute hemorrhagic diarrhea syndrome in dogs, but the onset of diarrhea caused by enterotoxin remains unknown. No gold standard exists for confirmation of C. perfringens enterotoxin in a feces of diarrheal dogs and for now diagnosis has been contingent on the simultaneous occurrence of clinical signs, the detection of large numbers of C. perfringens endospores in fecal smears and the immunodetection of CPE in fecal specimens by using an enzyme-linked immunosorbent assay (ELISA) together with the molecular polymerase chain reaction (PCR). The aim of this study was to establish the presence of C. perfringens enterotoxin in feces of healthy dogs and dogs with diarrhea using ELISA and PCR. Fecal samples were collected from 82 dogs, 40 of them were healthy and 42 were dogs with diarrhea of unknown etiology. Presence of C. perfringens enterotoxin (CPE) in 40% (17/42) dogs with diarrhea and 12.5% (5/40) healthy dogs was determined by ELISA test. The presence of CPE was not statistically significantly higher in dogs with diarrhea (p = 0.3179) than in healthy dogs. The prevalence of the cpe gene determined by the PCR method was 50% (21/42) in dogs with diarrhea and 70% (28/40) in healthy dogs. The presence of cpe gene was not statistically significantly higher in dogs with diarrhea compared to healthy dogs (p = 0.0903). Dogs with diarrhea were positive for CPE and cpe gene in 26% of cases (11/42) and in 10% (4/40) of healthy dogs. Statistical analysis showed a significant difference between the results obtained by the PCR method versus those obtained by the ELISA test (p = 0.0072). In conclusion, C. perfringens enterotoxin was present in both diarrheal and healthy dogs. Also, the results of this study have shown that a combination of PCR and ELISA methods is required for the reliable diagnosis of Clostridium perfringens enterotoxin in fecal samples.

• Keywords: Clostridium perfringens; enterotoksin; proljev; lančana reakcija polimerazom; imunoenzimni test
• Keywords (english): Clostridium perfringens; enterotoxin; enzyme-linked immunosorbent assay; polymerase chain reaction
• Language: croatian
• URN:NBN: urn:nbn:hr:178:615733
• Study programme: Title: Veterinary Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica veterinarske medicine (doktor/doktorica veterinarske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-01-28 09:49:01