Acaricides are hypothesized to reduce human risk of exposure to tick-borne pathogens by decreasing the abundance and/or infection prevalence of the ticks that serve as vectors for the pathogens. Acaricides targeted at reservoir hosts such as small mammals are expected to reduce infection prevalence in ticks by preventing their acquisition of zoonotic pathogens. By reducing tick abundance, reservoir-targeted or broadcast acaricides could reduce tick infection prevalence by interrupting transmission cycles between ticks and their hosts. Using an acaricide targeted at small-mammal hosts (TCS bait boxes) and one sprayed on low vegetation (Met52 fungal biocide), we tested the hypotheses that infection prevalence of blacklegged ticks with zoonotic pathogens would be more strongly diminished by TCS bait boxes, and that any effects of both acaricidal treatments would increase during the four years of deployment. We used a masked, placebo-controlled design in 24 residential neighborhoods in Dutchess County, New York. Analyzing prevalence of infection with <i>Borrelia burgdorferi</i>, <i>Anaplasma phagocytophilum</i>, and <i>Babesia microti</i> in 5380 nymphal <i>Ixodes scapularis</i> ticks, we found little support for either hypothesis. TCS bait boxes did not reduce infection prevalence with any of the three pathogens compared to placebo controls. Met52 was associated with lower infection prevalence with <i>B. burgdorferi</i> compared to placebo controls but had no effect on prevalence of infection with the other two pathogens. Although significant effects of year on infection prevalence of all three pathogens were detected, hypothesized cumulative reductions in prevalence were observed only for <i>B. burgdorferi</i>. We conclude that reservoir-targeted and broadcast acaricides might not generally disrupt pathogen transmission between reservoir hosts and tick vectors or reduce human risk of exposure to tick-borne pathogens.Published version