Preliminary Observations on the Density of Sporozoites of Theileria annulata

摘要: Hyalomma anatolicum heavily infected with Theileria annulata were fed on rabbits for 3 days. The ticks lightly ground in Eagle’s minimum essential medium (Gibco) Earle’s salts, NaHCO3 and HEPES supplemented bovine plasma albumin at 3.5% (MEM/BPA3.5) or 0.7% (MEM/BPA0.7, 1.016g/cm3 1.012g/cm3 respectively. suspensions centrifuged 100 × gav, 5 min. supernatant liquids then filtered through Millipore prefilters (AP25) cellulose ester filters (MF, 8μm 5μm pore, 47 mm 25 diameter). filtrates sporozoite tick debris. Self-forming density gradient media formulated. To 90 ml of pure Percoll (Pharmacia) was added 3.5g BPA. 10 10x concentrated MEM (without HEPES) 0.595g the pH adjusted to 7.45 7.5N NaOH. Percoll/BPA MEM/HEPES make stock isotonic (SIP/BPA3.5, approximately 7.1, 1.130g/cm3). Step-gradient formulated as above, but modified. 0.7g BPA added. 7.05 (SIP/BPA0.7, 7.35, 1.126g/cm3). final concentration both types SIP mM.