PurposeTheileriosis in cattle has a significant economic implication for dairy production globally. Thus, it is crucial to investigate the prevalence of bovine theileriosis, the causative agent and genotypes of Theileria species in dairy cattle in the Kurunegala District in the Intermediate zone, where the third largest population of dairy cattle in Sri Lanka is present and experienced a substantial reduction in the cattle population recently. MethodsSixty blood samples and background data were collected from three dairy farms in Galpokuna (n = 20), Koulwewa (n = 20), and Andigama (n = 20) areas. Haematocrit was used to identify anaemia while Giemsa-stained blood smears demonstrated the presence of piroplasms. A Fisher's Exact Test (p < 0.05) compared the prevalence of infection among age groups and farms. PCRs using species-specific primers designed to amplify regions of MPSP and 18 s rRNA genes followed by sequencing of selected amplicons allowed phylogeny of the species detected. ResultsAll three farms had semi-intensive farming practices keeping animals in a ‘closed’ setup with limited movement. Theileriosis prevalence in dairy cattle was 55.0 % with no difference among the farms (Koulwewa: 65.0 %, Galpokuna: 50.0 %, Andigama: 50.0 %). One-third of the cattle (33.3 %) were anaemic based on haematocrit results but most showed mild anaemia. Anaemia was significantly higher in calves (45.0 %) than in adults (25.0 %; χ2 = 5.743; p = 0.03) tested positive for theileriosis. None of the animals showed any other signs of theileriosis. PCR revealed the presence of both T. orientalis (pathogenic or benign) and T. annulata (pathogenic). The sequencing data revealed that the T. orientalis genotype present in Kurunegala District is type 7. ConclusionThis is the first report on T. orientalis and T. annulata in dairy cattle in the Intermediate zone of Sri Lanka. The type 7 of T. orientalis was more common showing severe anaemia in calves while mild anaemia or no anaemia in adults and sub-adults. In immunologically intact animals, the reported genotype of the parasite can persist asymptomatically for life, occasionally causing a relapse, particularly under stressful conditions like pregnancy, lactation, and rapid changes in environmental conditions. However, the susceptibility of calves for pathogenic and apathogenic genotypes of T. orientalis and the carrier status of asymptomatic animals needs further investigation.