The Clostridium perfringens enterotoxin from equine isolates; its characterization, sequence and role in foal diarrhoea.
Abstract: During a survey of foal diarrhoea between 1991 and 1994, Clostridium perfringens was significantly associated with disease with 56% of cases infected [1]. The contribution of enterotoxigenic C. perfringens to this association, was assessed by use of the reverse passive latex agglutination test for enterotoxin (RPLA; Oxoid Unipath) and vero cell toxicity neutralized by antitoxin on stored faecal samples and sporulated faecal isolates of C. perfringens. Polymerase chain reaction (PCR1) based on the DNA sequence for the whole enterotoxin gene [2] yielded a fragment from an equine isolate of the anticipated size which, cloned into plasmid M13 phage, had a sequence essentially identical to the published sequence. Consequently, all faecal isolates were also tested by PCR1 and for a part of the enterotoxin gene (PCR2). Significant association with diarrhoea (controls not in contact with cases) was found with positive RPLA tests on faeces (OR = 13, P = 0.002) and isolates (OR = 4.57, P = < 0.0001), vero cell toxicity of isolates (OR = 1.78, P = 0.026), and PCR1 (OR = nd, P = 0.029) but not PCR2 or vero cell toxicity of faeces. Significant association with diarrhoea was also found for isolates negative by RPLA (OR = 3.91; CI 2.05-7.57; P < 0.0001) or PCR1 (OR = 4.81; CI 2.84-8.20; P < 0.0001). Many of the isolates from RPLA positive faeces and verotoxic isolates were PCR negative and no evidence could be found for the presence of the enterotoxin gene in a random selection of RPLA positive/PCR negative isolates by gene probe on chromosomal DNA and PCR reaction product or vero cell toxicity neutralized by specific antiserum. Failure of the vero cell toxicity on faeces to be associated with diarrhoea or for cytotoxicity of cultures and RPLA on cultures to agree with the PCRs was believed to be related to the presence of other cytotoxins, the inherent cytotoxicity of equine faeces and to the poor specificity of the commercial antiserum used in the test. Enterotoxigenic C. perfringens could not account for the overall association of C. perfringens with foal diarrhoea because (a) cultures positive by PCR, RPLA or cytotoxicity were not significantly more common amongst isolates from cases than controls; and (b) the proportion of isolates from cases positive by PCR (PCR1 or PCR2) was too small at 9.7%.
Publication Date: 1998-05-21 PubMed ID: 9593490PubMed Central: PMC2809390DOI: 10.1017/s0950268897008534Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study examines the role of the Clostridium perfringens enterotoxin in foal diarrhea. Extensive tests were conducted to investigate the pathogen’s prevalence, potential for disease contribution, and the incidence of its enterotoxin gene in equine fecal samples.
Abstract Overview
This article focuses on a study conducted from 1991 to 1994, which investigated the significant association of Clostridium perfringens with foal diarrhoea, a prevalent health issue in young horses. In particular, the study strives to understand the contribution of enterotoxigenic C. perfringens – a specific type of this bacterium that produces an enterotoxin – to this disease.
Experimental Procedures
- The research team used diagnostic tests such as the reverse passive latex agglutination test (RPLA) and vero cell toxicity to detect the presence of the enterotoxin in stored fecal samples and bacterial isolates.
- Polymerase Chain Reaction (PCR), a technique used to amplify DNA, was also utilized to identify the presence of the enterotoxin gene in the bacteria collected from horses.
Findings
- Results showed a significant association between diarrhoea and positive RPLA tests on faeces and isolates, vero cell toxicity of isolates and PCR1 results.
- However, there were inconsistencies in the results. Isolates from RPLA-positive faeces and isolates showing vero cell toxicity were often PCR-negative, indicating an absence of the enterotoxin gene. This suggests that other factors could be at play aside from the involvement of the enterotoxin.
- Another finding was that the cytotoxicity of cultures and RPLA results on cultures did not always align with PCR outcomes, which was seen to be due to the presence of other cytotoxins, the inherent cytotoxicity of equine faeces, and the poor specificity of the commercial antiserum used in the test.
Conclusion
- The study concludes that enterotoxigenic C. perfringens might not be solely responsible for the overall connection of Clostridium perfringens with foal diarrhoea.
- This conclusion lies in two main points: cultures positive by PCR, RPLA or cytotoxicity were not significantly more common among the case bacterial isolates than controls, and the proportion of isolates from cases that tested positive by PCR was too small to be considered significant.
Cite This Article
APA
Netherwood T, Binns M, Townsend H, Wood JL, Mumford JA, Chanter N.
(1998).
The Clostridium perfringens enterotoxin from equine isolates; its characterization, sequence and role in foal diarrhoea.
Epidemiol Infect, 120(2), 193-200.
https://doi.org/10.1017/s0950268897008534 Publication
Researcher Affiliations
- Animal Health Trust, Newmarket, Suffolk.
MeSH Terms
- Animals
- Animals, Newborn
- Case-Control Studies
- Clostridium Infections / microbiology
- Clostridium Infections / veterinary
- Clostridium perfringens / classification
- Clostridium perfringens / genetics
- DNA, Bacterial / analysis
- Diarrhea / microbiology
- Diarrhea / veterinary
- Enterotoxins / genetics
- Horse Diseases / microbiology
- Horses
- Latex Fixation Tests
- Odds Ratio
- Polymerase Chain Reaction
- Reproducibility of Results
- Sequence Analysis, DNA
Citations
This article has been cited 5 times.- Khan MUZ, Humza M, Yang S, Alvi MA, Iqbal MZ, Zain-Ul-Fatima H, Khalid S, Munir T, Cai J. Occurrence and Toxicogenetic Profiling of Clostridium perfringens in Buffalo and Cattle: An Update from Pakistan.. Toxins (Basel) 2021 Mar 13;13(3).
- Mallicote M, House AM, Sanchez LC. A review of foal diarrhoea from birth to weaning.. Equine Vet Educ 2012 Apr;24(4):206-214.
- 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.
- 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.
- Netherwood T, Gilbert HJ, Parker DS, O'Donnell AG. Probiotics shown to change bacterial community structure in the avian gastrointestinal tract.. Appl Environ Microbiol 1999 Nov;65(11):5134-8.
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