Ticks are medically important and significant vectors of diseases affecting livestock, humans, and companion animals than any other arthropod vectors. In the absence of information on the relationship of tick species and piroplasms parasites in Lesotho, the current study was aimed at detecting piroplasms parasites of economic importance from ticks of domestic animals. A total of 322 pooled tick DNA samples were subjected to PCR screening for the presence of piroplasms. The overall infection rate of piroplasms was 7% with Babesia bigemina at 3.4% (11/322), B. bovis 0.3% (1/322), B. ovis 2.8% (9/322) and 0.6% (2/322) for B. motasi. DNA extracted from the Lesotho Rhipicephalus decoloratus and R. evertsi evertsi tested positive for the presence of B. bigemina with a 15% and 3% infection rate, respectively. Otobius megnini tested positive for only B. bovis at a 12.5% infection rate. Rhipicephalus e. evertsi was the only tick species PCR positive for ovine babesiosis with 3.2% for B. ovis and 0.7% for B. motasi. Equine piroplasm (Theileria equi and B. caballi) and Theileria (T. parva and T. ovis) parasites were not detected in the current study. The PCR-positive samples were confirmed by direct sequencing of the product. This study is the first to report on a relationship of Babesia parasites with tick species in Lesotho and it is evident that vector-borne diseases are present in ticks of domestic animals in this country. Research findings in this study require a joint effort from both veterinary and medical sectors to unite and conduct more epidemiological studies of tick-borne diseases in both animals and humans and to also determine the role played by tick species in the transmission of the detected parasites in domestic animals of Lesotho. This information provides a baseline knowledge of important piroplasms parasites and raising awareness of their prevalence in Lesotho.