To determine the presence and distribution of bovine theileriosis in the North Central region of Algeria, 358 DNA samples and 359 blood smears were analyzed from nine provinces. Theileria DNA extracted from cattle blood was amplified by fluorescence resonance energy transfer polymerase chain reaction (FRET-PCR). Blood smears were examined for Theileria piroplasms by microscopical examination (ME) of Giemsa-stained slides. While microscopical identification revealed only 42 animals being infected with Theileria piroplasms, PCR-positive amplification using Theileria genus-specific primers was obtained from 132 Theileria spp. (P < 0.0001). Among the 132 positives, 108 animals (81.8 %) were found positive of Theileria annulata, while 24 (18.2 %) were found positive for Theileria sp. (P < 0.0001). However, melting curve analysis of these latter samples revealed the presence of two different peaks, 51.5 ± 0.5 °C corresponding to Theileria sp1 and 52.5 ± 0.5 °C for Theileria sp2. Cloning and sequencing of Theileria sp1 and Theileria sp2 using the Cox primers indicated that these species are very closely related to Theileria buffeli. There is a highly significant difference in the distribution of theileriosis between different provinces (P < 0.0001). This disparity between provinces is probably due to differences in tick contact, influenced by the subhumid bioclimatic gradient and differences in agricultural land use.