Identification and Characterization of Anticoagulant Activities in the Saliva of the Lone Star Tick, Amblyomma americanum (L.)

Abstract

Anticoagulant activities against both the extrinsic and intrinsic coagulation pathways were identified in the saliva of partially fed female lone star ticks, Amblyomma americanum (L.). The activities of factor Xa and thrombin in the common pathway of the coagulation cascade were inhibited by tick saliva. The greatest anticoagulant activities were found in the saliva of ticks weighing more than 200 mg. The anticoagulant activities in tick saliva could be detected without preincubation of tick saliva with sheep plasma, but preincubation significantly increased the activities. Tick saliva anticoagulant activities were abolished by boiling for 15 min or being treated with trypsin for 1 hr. Phosphatidylcholine (3 mM) and phospholipase A2 inhibitor oleyloxyethyl phosphorylcholine (0.2 mM) did not affect the anticoagulant activities significantly, suggesting that the phospholipase A2 activity found in tick saliva does not contribute to the anticoagulant activities. Size exclusion high performance liquid chromatography revealed that the molecular weights of the anticoagulant activities were approximately 16,000 D. The anticoagulant activities in tick saliva are believed to play an important role in facilitating tick feeding by helping overcome the host hemostatic system.