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Molecular detection and prevalence of Theileria ovis and Anaplasma marginale in sheep blood samples collected from Layyah district in Punjab, Pakistan

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Abstract

Theileria ovis and Anaplasma marginale are intracellular pathogens affecting a wide range of animals, causing huge economic losses worldwide. The present study reports the molecular evidence of Theileria ovis and Anaplasma marginale in sheep blood samples (N = 218) collected from Layyah district in Punjab (Pakistan), where economy heavily relies on livestock. A 520 base pair fragment specific for 18S ribosomal RNA gene of Theileria ovis was PCR amplified in 23/218 (10.6%) sheep blood samples, while for Anaplasma marginale, a 265 base pair fragment specific for msp1b gene was generated in 15/218 (6.9%) sheep blood samples. Two blood samples were found co-infected (0.9%) with both parasites. Amplified PCR products of both parasites were confirmed by DNA sequencing and submitted to GenBank. Prevalence of both Theileria ovis (p = 0.3) and Anaplasma marginale (p = 0.4) varied non-significantly among the investigated sheep breeds. Tick burden on dogs present with sheep herds was found associated with Theileria ovis infection in sheep (p = 0.05). It was observed that lambs (p = 0.009), sheep in small herds (p = 0.04), and tick burden on dogs present with sheep herds (p = 0.01) were associated with Anaplasma marginale infection in sheep during the present study. In conclusion, we are reporting a higher prevalence of Theileria ovis than Anaplasma marginale in blood samples of sheep collected from Layyah district. Tick-infested dogs were found to be risk factors for the transmission of both pathogens in sheep, and tick control strategies should be extended to dogs associated with sheep herds in this area.

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All the epidemiological data associated with this project is presented in this manuscript.

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Authors and Affiliations

  1. Institute of, Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, Pakistan

    Kainat Abid, Sundas Bukhari, Muhammad Asif & Rehan Sadiq Shaikh

  2. Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan, Pakistan

    Ayesha Sattar, Maryam Arshad & Furhan Iqbal

  3. Department of Parasitology, Veterinary Faculty, University of Firat, 23119, Elazig, Turkey

    Munir Aktas & Sezayi Ozubek

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  1. Kainat Abid
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  2. Sundas Bukhari
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  3. Muhammad Asif
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  4. Ayesha Sattar
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  5. Maryam Arshad
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  6. Munir Aktas
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  7. Sezayi Ozubek
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  8. Rehan Sadiq Shaikh
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  9. Furhan Iqbal
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Contributions

FI and RSS had designed and supervised this study. KA, MA, and AS had collected blood and epidemiological data from sheep. KA, SB, AS, and MA had extracted DNA from the blood samples and performed PCRs. MA had purified PCR products and analyzed DNA sequences. MA and SO had constructed phylogenetic trees and analyzed the data. All these authors contributed in manuscript writing and approved the final version of the manuscript.

Corresponding authors

Correspondence to Rehan Sadiq Shaikh or Furhan Iqbal.

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Ethical Research Committee of the Institute of Molecular Biology and Biotechnology at Bahauddin Zakariya University Multan (Pakistan) approved all the experimental procedures and protocols applied in this study via letter number IMBB/Ethics/2019–23.

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Informed consent was obtained from livestock owners before including their animals in this study.

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Abid, K., Bukhari, S., Asif, M. et al. Molecular detection and prevalence of Theileria ovis and Anaplasma marginale in sheep blood samples collected from Layyah district in Punjab, Pakistan. Trop Anim Health Prod 53, 439 (2021). https://doi.org/10.1007/s11250-021-02870-5

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