Multiplex PCR for detection of Trypanosoma evansi and Theileria equi in equids of Punjab, India.
Abstract: Multiplex PCR for simultaneous detection of Trypanosoma evansi and Theileria equi in single-step reaction was optimized and employed on 108 equids (99 horses and 9 donkeys/mules) blood samples collected from two agro-climatic zones (Sub-mountain undulating zone and Undulating plain zone) of Punjab to evaluate the status of concurrent infection and associated risk factors. The amplification products of 257 and 709 bp targeting repetitive nucleotide sequence of variable surface glycoproteins of T. evansi and 18S rRNA gene of T. equi, respectively expressed high fidelity of the primer pairs with sequence homology to neighboring geographic isolates. The overall prevalence of T. evansi and T. equi was 3.7 and 1.85%, with Undulating plain zone at higher infection risk for T. equi (OR=3.24, 95% CI=0.28-83.65); and Sub-mountain undulating zone (OR=∞, 95% CI=0.25-∞) for T. evansi. Multiplex PCR revealed higher risk of infection of both T. equi (OR=6.75, 95% CI=0.58-175.38) and T. evansi (OR=2.11, 95% CI=0.05-80.36) in the farms with inappropriate management system. The risk factor associated with the type of host species had an odds ratio of 12.35 (95% CI=0.29-508.37) for donkeys/mules versus horses for T. evansi infection. This group was also at higher risk of infection with Odds ratio (OR) of 4 (95% CI=0.14-53.99) for T. equi. The current investigation brings out various commodities at risk of infection pertaining to equid trypanosomosis and theileriosis evaluated by a rapid and sensitive multiplex PCR assay.
Copyright © 2015 Elsevier B.V. All rights reserved.
Publication Date: 2015-05-29 PubMed ID: 26070973DOI: 10.1016/j.vetpar.2015.05.018Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research involved the development and use of a multiplex PCR test for simultaneous detection of Trypanosoma evansi and Theileria equi in blood samples taken from horses and donkeys/mules in Punjab, India. The researchers then assessed the risks and prevalence of infection in different geographic locations and management systems.
About the Research
- The research was conducted with the aim of simplifying and enhancing the diagnosis procedure of two infections in equids (horses, donkeys, and mules) – Trypanosoma evansi and Theileria equi.
- The research employed a multiplex PCR which was optimized for the simultaneous detection of both infections in a single-step reaction. This is advantageous in saving time, reducing the possibility of contamination, and saving costs associated with performing multiple tests.
- The research studied 108 equids (99 horses and 9 donkeys/mules) from two agro-climatic zones in Punjab, India: Sub-mountain undulating zone and Undulating plain zone.
Findings of the Research
- The multiplex PCR developed was successful in detecting the two infections with high accuracy. The primer pairs used were found to have significant sequence homology with geographic isolates.
- The overall prevalence of T. evansi was found to be 3.7% and T. equi was 1.85% in the sample studied.
- The Undulating plain zone was at a higher risk for T. equi infection, and the Sub-mountain undulating zone was at a higher risk for T. evansi.
- In the farms with inappropriate management systems, there was a higher risk of both T. evansi and T. equi infection.
- Donkeys and mules were found to be more susceptible to T. evansi infection than horses, demonstrating the varying risk factors related to different host species.
Implications of the Study
- This research provides a faster and more cost-effective method for the simultaneous detection of T. evansi and T. equi in equids.
- The study also identifies factors contributing to the spread of these infections among the equid population, which can guide decision-making in disease control measures.
- The study made it clear that better management practices could significantly reduce the spread of these infections.
- Understanding the risk factors associated with host species could help in formulating targeted interventions for disease control among different species of equids.
Cite This Article
APA
Sumbria D, Singla LD, Sharma A, Bal MS, Kumar S.
(2015).
Multiplex PCR for detection of Trypanosoma evansi and Theileria equi in equids of Punjab, India.
Vet Parasitol, 211(3-4), 293-299.
https://doi.org/10.1016/j.vetpar.2015.05.018 Publication
Researcher Affiliations
- Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, India.
- Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, India. Electronic address: ldsingla@gmail.com.
- Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, India.
- Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, India.
- National Research Centre on Equines, Haryana, India.
MeSH Terms
- Animals
- DNA, Protozoan
- Equidae
- India / epidemiology
- Multiplex Polymerase Chain Reaction / methods
- Multiplex Polymerase Chain Reaction / veterinary
- Odds Ratio
- Prevalence
- Risk Factors
- Species Specificity
- Theileria / isolation & purification
- Theileriasis / diagnosis
- Theileriasis / epidemiology
- Trypanosoma / isolation & purification
- Trypanosomiasis / diagnosis
- Trypanosomiasis / epidemiology
- Trypanosomiasis / veterinary
Citations
This article has been cited 4 times.- Torres R, Hurtado C, Pérez-Macchi S, Bittencourt P, Freschi C, de Mello VVC, Machado RZ, André MR, Müller A. Occurrence and Genetic Diversity of Babesia caballi and Theileria equi in Chilean Thoroughbred Racing Horses. Pathogens 2021 Jun 7;10(6).
- Metwally DM, Al-Turaiki IM, Altwaijry N, Alghamdi SQ, Alanazi AD. Molecular Identification of Trypanosoma evansi Isolated from Arabian Camels (Camelus dromedarius) in Riyadh and Al-Qassim, Saudi Arabia. Animals (Basel) 2021 Apr 17;11(4).
- Sumbria D, Das Singla L, Sharma A. Theileria equi and Babesia caballi infection of equids in Punjab, India: a serological and molecular survey. Trop Anim Health Prod 2016 Jan;48(1):45-52.
- Raftery AG, Gummery L, Garcia K, Mohite D, Capewell P, Sutton DGM. Equine trypanosomiasis, a systematic review and meta-analyses: Prevalence, morbidity and mortality. Equine Vet J 2026 Mar;58(2):291-319.
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