Ticks were processed individually throughout all procedures. The ticks were thawed and washed sequentially in 3% hydrogen peroxide, 100% isopropanol, and sterile distilled water for 1 min in each solution. The final distilled water wash was aspirated from the ticks and then the ticks were sectioned sagittally at midline with a sterile scalpel. Half of the tick was saved at −80 °C; the other half was used for DNA extraction. Briefly, 180 μl of ATL buffer (Qiagen, Valencia, CA, USA) and 20 μl of proteinase K were added to each tick and the ticks lysed overnight at 37 °C in an Eppendorf Thermomixer (Hauppauge, NY) with agitation at 1,400 rpm for 15 s every 15 min, before centrifuging the lysate for 3 min at 18,400× g. The supernatant was transferred into a sterile microfuge tube and DNA extracted using a Qiagen DNeasy Blood and Tissue kit in a Qiacube using the DNeasy Blood and Tissue protocol for Tissue and Rodent Tails (Qiagen, Valencia, CA, USA). Negative extraction controls consisted of sterile water processed via the same washing, chopping and extraction procedure used on the ticks.