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A Report on Molecular Detection and Phylogenetic Evaluation of Anaplasma marginale in Ticks and Blood Samples Collected from Cattle in District Layyah in Punjab (Pakistan)

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Abstract

Anaplasmosis is a tick-borne disease caused by obligate intercellular gram-negative bacteria, Anaplasma (A.) marginale. The present study reports on seasonal prevalence, epidemiology, and phylogeny of A. marginale in three cattle breeds from District Layyah, Southern Punjab, Pakistan. A total of 844 blood samples (Cross = 300, Holstein Friesian = 244, Sahiwal breed = 300) from apparently healthy cattle on seasonal basis were collected along with epidemiological data during May 2018 till April 2019. Polymerase chain reaction generated 265 base-pair amplicon specific for major surface protein-1b encoding gene of A. marginale in 8.6% (73/844) of enrolled cattle. The highest prevalence was observed during autumn (18.3%) followed by summer (9.7%) and winter season (7.1%). Holstein Friesian breed was most susceptible to A. marginale infection (13.1%) followed by Sahiwal (7.6%) and cross breed (6%). Representative amplified partial gene sequences of A. marginale were submitted to GenBank (Accession numbers MK032842 and MK032843). 37/844 (4.3%) Giemsa-stained blood smears were found positive for Anaplasma spp. Small number of ticks including Hyalomma anatolicum, Hyalomma excavatum, Rhipicephalus microplus, Haemaphysalis punctata were identified from cattle but none of them was found PCR positive for the presence of A. marginale. Analysis of epidemiological factors revealed that female cattle and farm with water supply from pool, farms where other dairy animals and dogs were living with cattle and dogs having ticks load on them had significant association with A. marginale prevalence. It was observed that white blood cell, lymphocytes (%), monocytes (%) hematocrit, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were significantly disturbed in A. marginale-positive than negative cattle.

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Funding

Funding was provided by Bahauddin Zakariya University (ORIC grant).

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Authors and Affiliations

  1. Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan, 60800, Pakistan

    Sehrish Ashraf, Asia Parveen & Furhan Iqbal

  2. Department of Pathobiology, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan

    Mian Muhammad Awais

  3. Institute of Molecular Biology and Biotechnology, University of Lahore, Lahore, Pakistan

    Quratulane Gillani

  4. Department of Parasitology, Fırat University, 23119, Elazig, Turkey

    Munir Aktas & Sezayi Ozubek

Authors
  1. Sehrish Ashraf
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  2. Asia Parveen
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  3. Mian Muhammad Awais
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  4. Quratulane Gillani
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  5. Munir Aktas
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  6. Sezayi Ozubek
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  7. Furhan Iqbal
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Contributions

FI has designed, supervised this project, and prepared the final manuscript. SA and AP have performed lab experiments and analyzed the data, MMA and QG have collected the blood samples and epidemiological data. While MA and SO have done DNA sequencing and phylogenetic analysis.

Corresponding author

Correspondence to Furhan Iqbal.

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The authors declare that they have no conflict of interest of any sort with anyone.

Ethical Approval

All the animal handling procedures and lab protocols were approved by the ethical committee of Institute of Pure and Applied Biology, Bahauddin Zakariya University Multan, Pakistan.

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Ashraf, S., Parveen, A., Muhammad Awais, M. et al. A Report on Molecular Detection and Phylogenetic Evaluation of Anaplasma marginale in Ticks and Blood Samples Collected from Cattle in District Layyah in Punjab (Pakistan). Curr Microbiol 78, 274–281 (2021). https://doi.org/10.1007/s00284-020-02256-0

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  • DOI: https://doi.org/10.1007/s00284-020-02256-0

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