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The blood parasites adversely affect the bovine population and are one of the major hindrances to productivity and health of livestock, globally. This study aimed to study the prevalence of some bovine hemoparasites, their molecular characterization, and associated risk factors in and around Bhubaneswar, a place in Eastern India. Microscopy (Giemsa stained thin blood smear) and polymerase chain reaction (PCR)-based prevalence studies were undertaken for a total of 106bovine blood samples belonging to different breed, age, and sex presented to Veterinary Clinical Complex. DNA was extracted by conventional method and primer targeting msp5 gene (382 bp) for  Anaplasma marginale,  Tams1 gene (572 bp) for  Theileria annulata  and VSG RoTat 1.2 gene (110 bp) for  Trypanosoma evansi , were used. Sequencing and phylogenetic analysis were also carried out for representative PCR products. The risk factors were correlated with prevalence by logistic regression analysis. Blood smear examination revealed a prevalence an overall of 48.11% (51/106) out of which the prevalence of  T. annulata ,  A. marginale  and  T. evansi,  and concurrent infection were reported to be 26.42%, 10.38%, 3.77%, and 7.55% respectively. All the samples found positive in microscopy were also found positive in PCR. PCR assay revealed an overall prevalence of 73.58% ( T. annulata : 37.74%,  A. marginale :17.92%,  T. evansi : 5.66% and concurrent infection:12.26%). A higher prevalence was observed in the age group of 2–4 years (37.74%), females (61.32%) and cross-bred Jersey (50%). Representative PCR products were sequenced and assigned by GenBank (OL550058:  A. marginale ; OL604429:  T. annulata  and OL550059:  T. evansi ). There is a paucity of disease mapping of these parasites in most parts of the country. The study is probably the first report of molecular characterization of  T. evansi  and  A. marginale  from the region which gives some information of possible genetic diversity of the isolates circulating in the region. Future research should include large-scale epidemiological surveys as well as concurrent diagnosis of these hemoparasites for better treatment and control strategies.

Molecular characterization and prevalence of bovine hemoprotozoan and rickettsial organism from Bhubaneswar, Eastern India