The overall prevalence rate of babesiosis on the basis of PCR assay ranged from 4.8 - 56.75% as per reports from different parts of India (Laha et al.,2013;Singh et al.,2014;Sarma et al.,2019) and 2.4 - 88.3% from around the globe (Bastos et al.,2004 ; Foldvari et al.2005 ; Porchet et al.,2007; Corali et al.,2018) . These differences might have occurred because of the differences in diagnostic techniques, the population sampled in the study, and climatic as well as managemental factors in locations where the research was conducted. A moderate rate of incidence observed during the present research might be due to the hot and humid environmental condition of the state, which favours the survival of tick vectors and a higher availability of stray dogs that act as transporting medium of the parasites due to their uncontrolled movements.
The prevalence of B.vogeli has been recorded in stray and pet dogs in north and south India ( Jain et al.,2018;Roopesh et al.,2018; Singla et al.,2016) , Europe(Cardoso et al., 2008;Ionita et al.,2012 ) Africa (M'ghirbi and Bouattour ,2008;Salem and Farag,2014). B.vogeli has also been incriminated as the commonest species in Latin America and Carribean (Panti-May and Rodriguez-Vivas,2020) . A higher presence of Babesia vogeli recorded in our study could be due to the dominating prevalence of Riphicephalus sanguineus, among canids of Bhubaneswar (Sahu et al.,2013). Riphicephalus sanguineus is attributed as natural vector for B.vogeli and H.canis (Dantas-Torres, 2008; Penzhorn, 2020). The present investigation also detected the presence of Hepatozoon canis along with Babesia sp, which corroborates with earlier reports from North east India (Sarma et al.,2019).This might be due to the primers Piro A1 and Piro B, which is specific to Babesia spp. but could also detect Hepatozoon species. The use of primers Babesia F and Babesia R by Oyamada et al.,2005 has also revealed cross-reaction between Babesia and Hepatozoon species.
Based on sequencing results and BLAST analysis, it was also observed that one sequence (KT246305) showed maximum homogeneity with sequences of B.gibsoni with a stray match with B.canis. In India, B.canis is yet to be reported in any molecular diagnosis possibly due to absence of potential vector. B.gibsoni infection in dogs has been reported earlier in blood smear examination from Bhubaneswar (Sahu et al.,2014) while molecular detection have been reported in different regions of India (Singh et al.,2014; Jain et al.,2018; Sarma et al.,2019). The molecular identification of B. gibsoni, B. vogeli and H.canis from canines of Punjab has been previously described (Singla et al., 2016). On phylogenetic analysis the present three isolates showed an affiliation with other B. vogeli isolates from different geographical regions. But one of the isolates showed no affiliation with other B. vogeli isolates though it was confirmed to be B. vogeli from Blast analysis, which might have originated from a different strain. B.gibsoni infected dogs exhibit varying clinical manifestation ranging from subclinical to fatal depending on the host body condition. Though B.vogeli is less pathogenic exhibiting moderate symptoms in adult dogs, they cause severe condition in puppies and splenectomised dogs (Wang et al., 2018).The study clearly re-established reliability of PCR as a technique over microscopy. Therefore, molecular diagnosis can facilitate pertinent treatment and control regimen.