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A Multi-copy Nucleic Acid-Based Diagnostic Test for Bovine Tropical Theileriosis

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Abstract

Background

Bovine tropical theileriosis (BTT) is a haemoprotozoan tick-borne disease that implicates huge losses to livestock in terms of considerable mortality and morbidity in tropical and subtropical regions of the globe. Currently available diagnostic methods have less specificity and sensitivity towards the detection of Theileria species. Therefore, an attempt was made to diagnose Theileria annulata by targeting a multi-copy gene, viz. mitochondrially encoded cytochrome b (MT-CYB) gene via polymerase chain reaction (PCR) in different agro-zones of India.

Methods and Results

129 cattle blood samples were collected from major livestock rearing regions of India and processed for both molecular and microscopic techniques. Screening of Giemsa-stained thin blood smears was able to detect 14 samples (10.85%) as positive for T. annulata. However, the MT-CYB gene-based PCR assay detected 107 samples (82.94%) positive for T. annulata out of 129 samples. Furthermore, the MT-CYB gene-based PCR assay was standardized in terms of its sensitivity and specificity. Specificity of PCR assay was evaluated against other common haemoprotozoan parasites of tropical countries viz. Babesia bigemina, Anaplasma marginale and Trypanosoma evansi. The multi-copy MT-CYB gene-based PCR assay provided an optimum level of sensitivity (up to the level of 10 femtogram) and high specificity. Haematological examination (Hb, PCV and TLC) of 113 samples revealed significantly (p < 0.05) decreased Hb and PCV levels in positive animals in comparison with the control group of healthy animals. However, the control group had significantly higher (p < 0.001) TLC levels than the positive group.

Conclusion

The MT-CYB gene-based PCR assay was found to be highly sensitive that can accurately detect the occurrence of T. annulata infection in carrier animals which are potential infection sources to healthier populations in naive demographic locations through infected ticks.

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All data are included in the manuscript.

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Acknowledgements

The authors pay gratitude to Director, ICAR-IVRI, Izatnagar for facilitating the study.

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Authors and Affiliations

  1. ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India

    Aquil Mohmad, B. C. Saravanan, Dinesh Chandra, Sheikh Firdous Ahmad, Waseem Akram Malla, Bilal Ahmad Malla & Nisha Bisht

  2. ICMR- National Institute of Research in Tribal Health, Jabalpur, Madhya Pradesh, 482003, India

    H. V. Manjunathachar

  3. Guru Angad Dev Veterinary and Animal Science University, Ludhiana, Punjab, 141004, India

    Ishfaq Maqbool

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  1. Aquil Mohmad
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  2. B. C. Saravanan
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  3. H. V. Manjunathachar
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  4. Dinesh Chandra
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Correspondence to Aquil Mohmad, B. C. Saravanan or H. V. Manjunathachar.

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No author has anything to declare regarding conflict of interest (financial or non-financial) with regards to the publication of this manuscript.

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All ethical considerations were taken into consideration during conduct of the experiment. The study did not involve any experimentation on animals except the collection of samples from field.

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Proper written consent was obtained from animal owners after explaining the procedure and aim of the study to them. Final diagnostic results were made accessible to the concerned owners for treatment aspect.

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Mohmad, A., Saravanan, B.C., Manjunathachar, H.V. et al. A Multi-copy Nucleic Acid-Based Diagnostic Test for Bovine Tropical Theileriosis . Acta Parasit. 67, 504–510 (2022). https://doi.org/10.1007/s11686-021-00428-x

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  • DOI: https://doi.org/10.1007/s11686-021-00428-x

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