Theileriosis is an important disease of large and small ruminants. The present study was undertaken to detect presence of Theileria spp. in dairy cattle and Hyalomma anatolicum in North East Iran using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). In this study, blood samples and H. anatolicum ticks were collected from 270 dairy cattle during the tick season of 2017-2018. The animals were investigated for presence of Theileria spp. by parasitological examination and subsequently by RLFP-PCR of blood. The H. anatolicum ticks collected were pooled according to their sex into groups of five. Tick salivary glands were dissected out in 0.85% saline under stereomicroscope. Presence of Theileria spp. in tick salivary glands were detected by PCR-RFLP. In this study, 4/270 (1.48%) blood smears were positive by microscopy. Theileria annulata was detected in 15 (5.5%) blood samples and in 4 (3.84%) tick salivary gland samples by PCR-RFLP. The results also showed H. anatolicum as the predominant tick vector associated with bovine theileriosis in North East Iran

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