High Throughput Tools for Tickborne Disease Surveillance and Investigation of Tick, Pathogen, and Commensal Microbiome Association at Single-Tick Resolution

Abstract

The prevalence of tickborne diseases worldwide is increasing virtually unchecked due to lack of effective control strategies. The transmission dynamics of tickborne pathogens are influenced by the tick microbiome, tick co-infection with other pathogens. Understanding this complex system could lead to new strategies for pathogen control, but will require large-scale, high-resolution data. Here we present a strategy that combines citizen science with new molecular strategies to provide the single-tick resolution data urgently needed to inform management of tickborne pathogens. Our citizen science-based initiative, Project Acari, harnessed the power of volunteers across the US to collect more than 3,000 ticks. To assay collected ticks, we developed a high-throughput screening method using Molecular Inversion Probes (MIPs) that identify tick species, associated pathogens, and the species on which the tick most recently fed. Applying MIPs to 853 individual ticks successfully identified the species of 715 ticks, of which 85 were infected with pathogens of 12 different species. We also detected host DNA in 60 ticks. We also generated the first comprehensive data on both prokaryotic and eukaryotic microbiome of individual ticks using full-length 16S and 18S sequencing. Our findings corroborate reports of the influence of tick species, sex, and geography on the tick prokaryotic microbiome. We also identify novel associations between the carriage of B. burgdorferi and specific microbial taxa. Our work underscores the power of citizen science, paired with high-throughput processing, to elucidate the ecology of tickborne disease and to guide pathogen-control initiatives.