Habitat preference and usage by disease vectors are directly correlated with landscapes often undergoing anthropogenic environmental change. A predominant type of land use change occurring in the United States is the expansion of native and non-native woody plant species in grasslands, but little is known regarding the impact of this expansion on regional vector-borne disease transmission. In this study, we focused on the impact of expanding eastern redcedar (Juniperus virginiana; ERC) and tested two hypotheses involving relationships between habitat preferences of adult tick species in rural habitats in central Oklahoma. Using CO₂ traps, we collected ticks from two densities of ERC and grassland and screened adult ticks for the presence of pathogen DNA. We found support for our first hypothesis with significantly more Amblyomma americanum (Linnaeus) and Dermacentor variabilis (Say) collected in ERC habitats than in grassland. Our second hypothesis was also supported, as Ehrlichia- and Rickettsia-infected A. americanum were significantly more likely to be collected from ERC habitats than grassland. As the first evidence that links woody plant encroachment with important tick-borne pathogens in the continental United States, these results have important ramifications involving human and companion animal risk for encountering pathogen-infected ticks in the southern Great Plains.