Anaplasmosis is endemic in Pakistan, and is economically one of the most important bovine diseases in Asia. It reduces livestock productivity and leads to high mortality especially in young animals. This study was aimed at identifying the potential risk factors responsible for the occurrence and spread of anaplasmosis in bovine. A total of 900 (cattle = 479, buffalo = 421) blood samples were collected irrespective of age and sex from 3 distinct temporal zones of Khyber Pakhtunkhhwa (KPK) province of Pakistan. Polymerase chain reaction (PCR) technique was used for the molecular detection of anaplasmosis in comparison with microscopy of stained blood smears to compare test sensitivity for its diagnosis. Data collected on a piloted questionnaire including 11 predicting variables which were analyzed using R-statistical software, and association between the dependent and independent variables was assessed using univaraible analysis. Automated and manual approaches were exercised, producing comparable models. Key risk factors identified in all the approaches included specie of the animal, breed of animal, sex of animal, tick infestation status, previous tick history, tick control status, and acaricides use (Odds ratio > 1). Also, 165 samples produced the 611bp DNA fragment specific for 16SrRNA gene of Anaplasma spp. The samples were confirmed for anaplasmosis through sequencing and BLAST queries. The findings of the current study are in line with the results of previous studies conducted in other countries identifying similar potential risk factors for anaplasmosis spread in bovine. Enhancing the protective measures for controlling the identified risk factors can reduce the spread of anaplasmosis in Pakistan.