Genetic Diversity of Canine Babesia Species Prevalent in Pet Dogs of Punjab, Pakistan

Simple Summary Babesiosis is caused by the intra-erythrocytic Babesia species in dogs. Different species are reported worldwide. The present study was conducted on pet dogs of two districts of Punjab, Pakistan, including Lahore and Narowal. Conventional microscopic and molecular tests, including conventional and nested PCR, were used for identification of diseased dogs and prevalent species. About 42% of dogs were tested positive for babesiosis and only two species including Babesia canis and gibsoni were prevalent. Canine babesiosis is a serious threat to dogs' health worldwide, caused by the intra-erythrocytic Babesia species. The present study was carried out in pet dogs presented at three clinics of Lahore and one of Narowal in Punjab, Pakistan. Two hundred blood samples (50 from each clinic) were collected and screened by microscopy for Babesia spp. Out of 200 samples, 84 (42%) were found to be positive for babesiosis. The highest number of positive cases (50%) was recorded in dogs at Narowal clinic. Non-significant variation (p > 0.05) was observed in the prevalence of babesiosis in dogs in relation to sex and age. Positive samples were further confirmed by a polymerase chain reaction using 18S-rRNA genus-specific and species-specific primers. Amplicons were further analyzed by nucleotide sequencing for genetic diversity. Babesia canis and gibsoni were confirmed by genome sequencing in all diseased dogs. These isolates closely resembled each other, but differed from previous reported strains. In conclusion, pet dogs suffering from babesiosis were infected with B. canis and gibsoni, while in other countries, other Babesia species are also prevalent.