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Recently, a two-year study found that long-term prescribed fire significantly reduced tick abundance at sites with varying burn regimes (burned surrounded by burned areas [BB], burned surrounded by unburned areas [BUB], and unburned surrounded by burned areas [UBB]). In the current study, these ticks were tested for pathogens to more directly investigate the impacts of long-term prescribed burning on human disease risk. A total of 5,103 ticks (4,607 Amblyomma americanum, 76 Amblyomma maculatum, 383 Ixodes scapularis, two Ixodes brunneus, and 35 Dermacentor variabilis) were tested for Borrelia spp., Rickettsia spp., Ehrlichia spp., and Anaplasma phagocytophilum. Long-term prescribed fire did not significantly impact pathogen prevalence except that A. americanum from burned habitats had significantly lower prevalence of Rickettsia (8.7% and 4.6% for BUB and UBB sites, respectively) compared to ticks from control sites (unburned, surrounded by unburned [UBUB])(14.6%). However, during the warm season (spring/summer), encounter rates with ticks infected with pathogenic bacteria was significantly lower (98%) at burned sites than at UBUB sites. Thus, despite there being no differences in pathogen prevalence between burned and UBUB sites, risk of pathogen transmission is lower at sites subjected to long-term burning due to lower encounter rates with infected ticks.