Molecular detection of Theileria annulata infection in cattle by conventional PCR and quantitative real time PCR in Indi
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ORIGINAL ARTICLE
Molecular detection of Theileria annulata infection in cattle by conventional PCR and quantitative real time PCR in India Ahmed Magdy Selim1,2 • Manoranjan Das1 • Santosh Kumar Senapati1 • Geeta Rani Jena1 • Chinmoy Mishra3 • Indramani Nath4 • Shantibhusan Senapati5 Manisha Sethi5
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Received: 25 July 2020 / Accepted: 18 September 2020 Ó Indian Society for Parasitology 2020
Abstract Theileria annulata (T. annulata) is a tick-borne apicomplexan parasite that affects bovine. It is endemic in many tropical and subtropics areas, including Odisha, India. The objective of this study is to identify T. annulata infection in the peripheral blood of cattle as a biological sample by conventional PCR (cPCR) and quantitative PCR (qPCR). The phylogenetic analysis was done using the T. annulata merozoite surface antigen (Tams 1) gene. Out of 552 samples of examined blood smears by microscopy, 454 (82.24%) animals were positive for Theileria species. Out of 454 samples, 96 samples were further examined by both cPCR and qPCR, 52 samples (54.16%) were found positive for T. annulata in both PCR methodologies. Phylogenetic
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12639-020-01278-6) contains supplementary material, which is available to authorized users. & Ahmed Magdy Selim [email protected] 1
Department of Clinical Medicine, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha 751003, India
2
Internal Medicine, Infectious Diseases and Fish Diseases Department, Faculty of Veterinary Medicine, Mansoura University, 35516 Mansoura, Egypt
3
Department of Animal Breeding and Genetics, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha 751003, India
4
Department of Veterinary Surgery, College of Veterinary Science and Animal Husbandry, Odisha University of Agriculture and Technology, Bhubaneshwar, Odisha 751003, India
5
Institute of Life Sciences, Bhubaneswar, Odisha, India
analysis revealed that T. annulata Odisha isolate was closely related to T. annulata Uttarakhand, India isolate (KM061799) and Hyderabad, India isolate (MK034702) with Nucleotide sequence identity 95.36%, 95.25%, respectively. This is the first study to detect T. annulata by qPCR in Odisha and supported that both PCR techniques were equally effective for the detection of Tams 1 gene of T. annulata in cattle’s blood. Keywords Blood Cattle PCR Theileria annulata
Introduction Theileriosis is a tick-borne parasitic disease caused by hemoprotozoan parasites, which commonly infect ruminants especially cattle in tropical and subtropical regions (Nazifi et al. 2009; Demessie and Derso 2015). Theileria is an intracellular protozoan parasite transmitted by ticks and has complex life cycles in vertebrate and invertebrate hosts together (Sitotaw et al. 2014). The presence of infective ticks around the year and transplacental transmission of T. a
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