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A study on prevalence and molecular characterization of trypanosomal species infecting equines in Lahore region, Pakistan

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Abstract

Trypanosomiasis is an important protozoal disease with a diverse range of susceptible host including human. In the current study, molecular characterization of prevalent species was done through a pan-trypanosome polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP). A total of three hundred (n = 300) equines including horses, donkeys and mules (100 each) were randomly selected and the equine blood samples were subjected to screening for trypanosomes through microhaematocrit centrifuge technique (MHCT), conventional PCR, semi-nested PCR and RFLP. Overall prevalence of trypanosomal species was 8% (24/300) as revealed by MHCT and species wise prevalence in horses, donkeys and mules was 4.33% (13/300), 1.33% (4/300) and 2.33% (7/300), respectively. Conventional and semi-nested PCR depicted an overall prevalence of 21% (63/300) and species wise prevalence in horses, donkeys and mules was 12% (36/300), 3.67% (11/300) and 5.33% (16/300), respectively. RFLP analysis of the semi-nested products, using Msp1 and Eco571 enzymes, negated the presence of T. congolense, T. brucei, T. vivax, T. theileri, and T. vivax in the positive samples and revealed that the animals might be suffering from T. evansi infection as the enzymes used were not able to detect this species. This hypothesis was further confirmed by using T. evansi specific primers which depicted all of the 63 samples were positive for T. evansi. It is inferred that T. evansi is the major trypanosome species prevalent in equines. Furthermore, PCR is more sensitive as compared to microscopic examination and the pan-trypanosome PCR–RFLP assay is suitable for carrying out laboratory diagnosis of field samples and epidemiological studies. Further studies on the possibilities of use of other restriction enzymes may help to improve the species specificity of the assay.

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Acknowledgements

The authors highly acknowledge the generous funding by Higher Education Commission, Islamabad, Pakistan and also acknowledge the cooperation extended by the administrative and technical staff of the Outdoor Veterinary Hospital, UVAS, Society for the Prevention of Cruelty to Animals (SPCA), and Brooke’s Hospital of Lahore for assistance in sample collection.

Authors’ contribution

NS conducted the experiments, ZIC supervised the experiments, AA and KM were co-supervisors, MS and IA helped in experiments, AH and SAK reviewed the manuscript before submission.

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Authors and Affiliations

  1. College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China

    Naveed Sabir & Muhammad Shahid

  2. Faculty of Veterinary Science, University of Veterinary and Animal Sciences (UVAS), Out Fall Road, Lahore, 54000, Pakistan

    Zafar Iqbal Chaudhry, Asim Aslam, Khushi Muhammad & Ishtiaq Ahmad

  3. Faculty of Veterinary and Animal Science, The University of Poonch Rawalakot, Rawalakot, 12350, Pakistan

    Abid Hussain & Shahzad Akbar Khan

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  1. Naveed Sabir
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  2. Zafar Iqbal Chaudhry
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  3. Asim Aslam
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  4. Khushi Muhammad
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  5. Muhammad Shahid
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  6. Abid Hussain
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  7. Shahzad Akbar Khan
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  8. Ishtiaq Ahmad
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Correspondence to Naveed Sabir.

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The authors declare that they have no conflict of interest.

Ethical approval

The experimental design and protocol was submitted to the directorate of advanced studies and research, UVAS, Lahore and was duly approved before the start of the study.

Informed consent

Informed consent was obtained from all of the owners of the animals included in the study.

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Sabir, N., Chaudhry, Z.I., Aslam, A. et al. A study on prevalence and molecular characterization of trypanosomal species infecting equines in Lahore region, Pakistan. J Parasit Dis 42, 96–101 (2018). https://doi.org/10.1007/s12639-017-0972-9

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  • DOI: https://doi.org/10.1007/s12639-017-0972-9

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