For the bacterial community analysis, a segment of the conserved bacterial 16S rRNA gene was amplified from the individual tick DNA extractions using universal primers 515F and 806R that flank the V4 region (Caporaso et al., 2012). The 806R primers also contained a unique 12-nucleotide Golay “barcode” for each sample that allowed us to pool the PCR products from all the samples into one Illumina MiSeq sequencing run but then to identify sequences derived from each individual tick. PCR reactions were conducted in a total volume of 40 μL using Taq98® Hot Start 2X Master Mix (Lucigen, Middleton, WI, USA) with primer concentrations at 0.2 μM. PCR cycling conditions were: denaturation at 98 °C for 2 min followed by 35 cycles of 98 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; then final extension at 72 °C for 10 min before holding the PCR products at 4 °C. The PCR products were visualized under UV light on 1% agarose gels stained with ethidium bromide before being normalized and sequenced on an Illumina MiSeq instrument by The Scripps Research Institute DNA Array Core Facility using their standard protocols (TSRI, San Diego, CA, USA).