Molecular analysis of Clostridium difficile isolates recovered from horses with diarrhea.
Abstract: Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A(-)B(+) variants. Thirteen of forty-eight isolates (27.0%) did not posses any toxin genes (A(-)B(-)CDT(-)). A positive EIA result was reported in 17 (44%) of the cases. There was no association between the presence of different ribotypes or strains and toxin gene(s) profiles and the clinical outcome.
Publication Date: 2006-10-20 PubMed ID: 17112686DOI: 10.1016/j.vetmic.2006.10.013Google Scholar: Lookup
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- Journal Article
Summary
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This research studied the characteristics of a bacteria called Clostridium difficile, which causes diarrhea in horses, by examining its molecular properties in 48 different samples. The study also tested horse feces for the presence of this bacteria’s toxins and found no correlation between variations in bacteria strain or toxin profiles and the horse’s clinical outcome.
Methodology and Findings
- In the study, the researchers analyzed the molecular characteristics of 48 Clostridium difficile isolates. These were obtained from horses that had diarrhea and were admitted to a veterinary hospital.
- The analysis involved PCR-ribotyping and identifying the toxin gene profile. PCR-ribotyping is a method used to classify different strains of bacteria, while the toxin gene profile identifies the specific toxins produced by the bacteria.
- The researchers also tested horse feces for the presence of Clostridium difficile toxin A and B using an Enzyme Immunosorbent Assay (EIA). This is a test that detects and measures antibodies in the blood. It can be used to determine if the patient has been exposed to a particular antigen – in this case, the clostridium difficile toxins.
Results and Conclusion
- Out of the 48 strains studied, they found 27 strains (56.25%) had the tcdA toxin gene, 35 strains (72.91%) had the tcdB toxin gene and 2 strains (4.1%) had the cdtB toxin gene.
- A total of 8 isolates (16.6%) were identified as A(-)B(+) variants, meaning they don’t produce toxin A, but they produce toxin B.
- Thirteen (27%) of the isolates didn’t possess any toxin genes, marked as A(-)B(-)CDT(-).
- The EIA test showed positive results in 17 cases (44%) indicating the presence of either toxin A or B.
- The study found there was no association between different bacterial strains or their toxin gene profiles and the clinical outcomes in horses. This implies that the severity or type of disease is not necessarily dependent on the specific strain or toxins produced.
Cite This Article
APA
Arroyo LG, Staempfli H, Weese JS.
(2006).
Molecular analysis of Clostridium difficile isolates recovered from horses with diarrhea.
Vet Microbiol, 120(1-2), 179-183.
https://doi.org/10.1016/j.vetmic.2006.10.013 Publication
Researcher Affiliations
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ont., Canada N1G 2W1. larroyo@uoguelph.ca
MeSH Terms
- ADP Ribose Transferases / genetics
- Animals
- Bacterial Proteins / genetics
- Bacterial Toxins / genetics
- Clostridioides difficile / classification
- Clostridioides difficile / genetics
- Clostridioides difficile / isolation & purification
- Diarrhea / microbiology
- Diarrhea / veterinary
- Enterocolitis, Pseudomembranous / microbiology
- Enterocolitis, Pseudomembranous / mortality
- Enterocolitis, Pseudomembranous / veterinary
- Enterotoxins / genetics
- Feces / microbiology
- Horse Diseases / microbiology
- Horse Diseases / mortality
- Horses
- Ribotyping / veterinary
Citations
This article has been cited 14 times.- Uchida-Fujii E, Niwa H, Senoh M, Kato H, Kinoshita Y, Mita H, Ueno T. Clostridioides difficile infection in thoroughbred horses in Japan from 2010 to 2021. Sci Rep 2023 Aug 11;13(1):13099.
- 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.
- Riddle DJ Dr, Dubberke ER Dr. Trends in Clostridium difficile Disease: Epidemiology and Intervention. Infect Med 2009;26(7):211-220.
- Silva RO, Santos RL, Pires PS, Pereira LC, Pereira ST, Duarte MC, de Assis RA, Lobato FC. Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil. Braz J Microbiol 2013;44(1):133-7.
- Yan W, Shin KS, Wang SJ, Xiang H, Divers T, McDonough S, Bowman J, Rowlands A, Akey B, Mohamed H, Chang YF. Equine hyperimmune serum protects mice against Clostridium difficile spore challenge. J Vet Sci 2014;15(2):249-58.
- Schoster A, Arroyo LG, Staempfli HR, Shewen PE, Weese JS. Presence and molecular characterization of Clostridium difficile and Clostridium perfringens in intestinal compartments of healthy horses. BMC Vet Res 2012 Jun 29;8:94.
- Botha M, Botes M, Loos B, Smith C, Dicks LM. Lactobacillus equigenerosi strain Le1 invades equine epithelial cells. Appl Environ Microbiol 2012 Jun;78(12):4248-55.
- Keessen EC, Hopman NE, van Leengoed LA, van Asten AJ, Hermanus C, Kuijper EJ, Lipman LJ. Evaluation of four different diagnostic tests to detect Clostridium difficile in piglets. J Clin Microbiol 2011 May;49(5):1816-21.
- Metcalf D, Reid-Smith RJ, Avery BP, Weese JS. Prevalence of Clostridium difficile in retail pork. Can Vet J 2010 Aug;51(8):873-6.
- Clements AC, Magalhães RJ, Tatem AJ, Paterson DL, Riley TV. Clostridium difficile PCR ribotype 027: assessing the risks of further worldwide spread. Lancet Infect Dis 2010 Jun;10(6):395-404.
- Petry S, Tapprest J, Maillard K, Barbut F, Duquesne F, Kozak S, Foucher N, Bernez-Romand M, Bridoux L, Poquet I. Clostridioides difficile in equidae necropsied in Northwestern France, between 2019 and 2021. Microbiol Spectr 2026 Feb 3;14(2):e0216525.
- Borges AS, Zakia LS, Yu S, Surette MG, Arroyo LG. Isolation of Clostridioides difficile from a Large Animal Veterinary Teaching Hospital Environment. Animals (Basel) 2025 Sep 15;15(18).
- Hain-Saunders NMR, Knight DR, Harvey A, Bruce M, Hampson BA, Riley TV. Clostridioides difficile in feral horse populations in Australia. Appl Environ Microbiol 2025 May 21;91(5):e0211424.
- Haywood LMB, Sheahan BJ. A Review of Epithelial Ion Transporters and Their Roles in Equine Infectious Colitis. Vet Sci 2024 Oct 7;11(10).
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