THEILERIA LESTOQUARDI - MATURATION AND QUANTIFICATION IN HYALOMMA-ANATOLICUM-ANATOLICUM TICKS

The maturation and quantification of Theileria lestoquardi (T. hirci) parasites in unfed and partially fed adult Hyalomma anatolicum anatoli cum ticks was studied using (1) methyl green pyronin (MGP) staining of salivary glands, (2) in vitro infection of peripheral blood mononucle ar cells (PBM) with parasites harvested from infected ticks and (3) a semiquantitative polymerase chain reaction (PCR). With MGP staining th e greatest infection rate was seen in unfed ticks. Feeding resulted in a gradual reduction in the number of infected acini with a concomitan t increase in the maturity of the parasites. In vitro infection of she ep PBM with titrated ground-up tick supernate (GUTS) demonstrated that infectivity peaked between 2 and 4 days of tick feeding whereas GUTS prepared from unfed ticks was not infective. The polymerase chain reac tion (PCR) was both sensitive and specific, detecting T. lestoquardi D NA in unfed and partially fed ticks, with a maximum sensitivity of 0.0 22 infected acinus/tick in 2-day fed ticks, though it gave no indicati on of the infectivity of the parasite.