English
Use of a real-time polymerase chain reaction-based fluorogenic 5′ nuclease assay to evaluate insect vectors of Corynebacterium pseudotuberculosis infections in horses.
Abstract: To develop and use a sensitive molecular assay for detecting the phospholipase D (PLD) exotoxin gene of Corynebacterium pseudotuberculosis in an attempt to identify insect vectors that may be important in transmission of clinical disease in horses. Methods: 2,621 flies of various species. Methods: A real-time polymerase chain reaction (PCR)-based fluorogenic 5' nuclease (TaqMan) system (ie, TaqMan PCR assay) was developed for the detection of the PLD gene in insects. Flies were collected monthly (May to November 2002) from 5 farms in northern California where C. pseudotuberculosis infection in horses is endemic. Three of the 5 farms (which housed a total of 358 horses) had diseased horses during the study period. A total of 2,621 flies of various species were tested for the PLD gene of C. pseudotuberculosis. Results: Evidence of bacterial DNA for the PLD gene was detected in skin biopsy specimens from clinically affected horses and from 3 fly species collected from farms where affected horses were housed. Farms with a high incidence of diseased horses had a high proportion of insects carrying the organism. High percentages of flies with positive results for the PLD gene were observed in October, when most clinically affected horses were observed. Conclusions: Our results are consistent with the hypothesis that C. pseudotuberculosis may be vectored to horses by flies. Three potential vectors were identified, including Haematobia irritans, Stomoxys calcitrans, and Musca domestica. The organism can be identified in up to 20% of house flies (Musca domestica) in the vicinity of diseased horses.
Publication Date: 2004-06-17
PubMed ID: 15198224DOI: 10.2460/ajvr.2004.65.829Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article is about the development of a molecular test to identify the bacteria Corynebacterium pseudotuberculosis in flies, which are suspected to carry and transmit the disease to horses. By using this test, the researchers were able to examine potential insect vectors, their presence on farms with infected horses, and the timing of infections.
Methodology
- The researchers used a method known as the real-time polymerase chain reaction (PCR)-based fluorogenic 5′ nuclease (TaqMan) system, also known as the TaqMan PCR assay. This assay was developed specifically for detecting the phospholipase D (PLD) exotoxin gene of C. pseudotuberculosis in insects.
- The flies were collected monthly from May to November 2002 from five different farms in northern California where C. pseudotuberculosis infection in horses is endemic.
- Three of these farms, housing 358 horses in total, had reported sick horses during the study period. Overall, 2,621 flies of various species were tested for the presence of the PLD gene.
Results
- The researchers were able to detect the bacterial DNA for the PLD gene in skin biopsy samples from affected horses and from three species of flies collected from farms with infected horses. The high presence of bacterial DNA in these flies suggets they act as vectors in the spreading of the disease.
- The study also revealed that farms with a high rate of infection in horses also had a high proportion of insects carrying the bacteria.
- The researchers also reported that high percentages of flies with positive results for the PLD gene were found in October, the same time period when most clinically affected horses were observed.
Conclusion
- The results of this study support the hypothesis that C. pseudotuberculosis is likely transmitted to horses by flies.
- The three potential vectors that were identified include Haematobia irritans, Stomoxys calcitrans, and Musca domestica, common species of flies.
- In particular, it was noted that up to 20% of house flies (Musca domestica) in proximity to diseased horses could be identified with the organism, suggesting they play a significant role in disease transmission.
Cite This Article
APA
Spier SJ, Leutenegger CM, Carroll SP, Loye JE, Pusterla JB, Carpenter TE, Mihalyi JE, Madigan JE.
(2004).
Use of a real-time polymerase chain reaction-based fluorogenic 5′ nuclease assay to evaluate insect vectors of Corynebacterium pseudotuberculosis infections in horses.
Am J Vet Res, 65(6), 829-834.
https://doi.org/10.2460/ajvr.2004.65.829
Publication
Researcher Affiliations
- Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
MeSH Terms
- Animals
- California
- Corynebacterium Infections / microbiology
- Corynebacterium Infections / veterinary
- Corynebacterium pseudotuberculosis / genetics
- DNA Primers
- Diptera / microbiology
- Genetic Testing
- Horse Diseases / microbiology
- Horses
- Insect Vectors / microbiology
- Phospholipase D / genetics
- Reverse Transcriptase Polymerase Chain Reaction
Citations
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