Human ehrlichiosis is the second most common tick-borne disease reported in Tennessee after Rocky Mountain spotted fever. Two closely related ehrlichiae, Ehrlichia chaffeensis and Ehrlichia ewingii, are both causative agents of human disease and are transmitted by Amblyomma americanum, the lone star tick. Prevalence rates and distribution patterns of these pathogens among ticks in Tennessee are currently unknown. To understand prevalence and exposure risk of Ehrlichia spp., we tested 616 ticks (309 Amblyomma americanum (L.), 277 Dermacentor variabilis (Say), 17 Ixodes texanus (Banks), 7 Ixodes cookei Packard, 4 Ixodes scapularis (Say), and 2 Amblyomma maculatum Koch) from 46 counties for E. chaffeensis and 324 ticks (238 A. americanum and 86 D. variabilis) from 29 counties for E. ewingii. Overall, E. chaffeensis was detected in 2.6% (8/309) of A. americanum and E. ewingii in 0.8% (2/238). Ehrlichia spp. DNA was not detected in any tick species other than A. americanum. Although sample sizes were low in many counties, all positive ticks were identified in the Interior Plateau and Southeastern Plains ecoregions which is where the majority of human ehrlichiosis cases are reported from Tennessee (e.g., 66.3% of the human cases in 2008 are from the Interior Plateau ecoregion). The data from this pathogen survey combined with frequent human case reports from certain areas indicate potential "hot spots" for ehrlichiosis infection. Targeted vector control interventions in these areas may help decrease human ehrlichiosis transmission.