Babesiosis is endemic in Pakistan and is one of the most economically important bovine diseases that cause huge economic losses and high mortality in young animals. An epidemiological study was conducted to unveil the prevalence and associated risk factors of Babesia bovis (B. bovis) in distinct climatic regions. A total of 900 (cattle=479, buffaloes=421) blood samples were collected from three distinct temporal zones of Khyber Pakhtunkhwa (KPK) province, Pakistan. The samples were analyzed by polymerase chain reaction (PCR) amplifying spherical body protein-4 (BbSBP-4) gene. Chi-square test, univariate analysis and multivariate logistic regression were used to analyze the data. The overall, prevalence in three distinct temporal zones of KPK province was found to be 10.11%. A higher prevalence of B. bovis was recorded in cattle 11.90%, compared with buffaloes 8.08% (OR:1.537, CI:0.984-2.403). Sequencing and phylogenetic analysis of locally isolated B. bovis showed sequence homology with the reported Syrian strain using NCBI BLAST tool. Species of the animal, sex of animals, tick infestation status, previous tick history, and tick control status, management type and geo-location were the significant (OR>1) risk factors associated with the occurrence. This study is the first molecular evidence of B. bovis and its associated risk factors in climatically distinct regions of KPK province, Pakistan.

Key words: Bovine, PCR, Spherical body protein-4, Temporal zones