Coexistence of Multiple Theileria annulata Genotypes Circulating in Neonatal Calves in Semi-arid India

Abstract

Background

Knowledge of local isolates and strains is a prerequisite for the development of either effective mass vaccination strategy or a suitable molecular marker-based diagnostic tool.

Purpose

The pathogenesis of Bovine tropical theileriosis (BTT), caused by Theileria annulata in susceptible ruminants, is known to vary depending upon the nature of isolate and strain involved. Therefore, RFLP and sequencing-based characterization of Indian isolates of T. annulata were attempted using TAMS gene.

Method

In the present study, TAMS 1 gene of T. annulata was amplified from 25 naturally infected calves from the BTT endemic semi-arid zone of Northern India. The amplified products were then digested with three restrictions enzymes viz., Taq I, Rsa I, and Alu I to find out the variations in pattern of restriction digests, so as to have an idea about the various isolates of T. annulata present in the studied area. Around 14 samples covering all the variants (from the PCR–RFLP patterns) were sequenced and submitted in NCBI (MH277607–MH277620).

Result

Coexistence of 4 variant genotypes was detected upon in-silico analysis of RFLP and sequence variations.

Conclusion

The nucleotide variations alongside the chromatogram analysis revealed point mutations leading to presence of noticeable genetic diversity among the isolates.

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Acknowledgements

The authors are highly thankful to the Director Research, Dean Biotechnology, Dean CoVSc, and Vice Chancellor, DUVASU, for the facilities provided. The authors also want to acknowledge the various funding agencies like Indian Council of Agricultural Research (ICAR) and Rashtriya Krishi Vikas Yojana (RKVY) for sanctioning various projects to the University for the Procurement of Instruments to carry out such work. The authors also acknowledge Incharge, Central Instrumentation Facility (CIF) for permitting the use of instruments required in the study.

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Correspondence to Vikrant Sudan.

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The authors declare that they have no conflict of interest.

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The blood was collected from the animals in compliance with the ethical standards and guidelines of the Institutional Animal Ethics Committee (IAEC) and permission was granted via. order no IAEC/17/23.

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Paliwal, S., Sudan, V., Shanker, D. et al. Coexistence of Multiple Theileria annulata Genotypes Circulating in Neonatal Calves in Semi-arid India. Acta Parasit. 65, 679–685 (2020). https://doi.org/10.2478/s11686-020-00196-0

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Keywords

  • Genetic variations
  • TAMS 1 gene
  • Theileria annulata
  • PCR–RFLP
  • Sequencing