Theileria-free Hyalomma anatolicum larvae were fed on a naturally infected sheep with Theileria lestoquardi. Resulting flat nymphs of the tick were able to transmit T. lestoquardi infection upon feeding to 3/3 susceptible sheep. Adults emerging from the same batch of larvae were also infective to 3/3 susceptible sheep when they had the infection during their larval feeding. Transmission of T. lestoquardi to sheep was confirmed through clinical monitoring, examination of blood and lymph node biopsy smears, serology using indirect immunoflourescent test, and molecular using polymerase chain reaction technique.