The southeastern United States is experiencing rapid environmental changes associated with urbanization. An ongoing question is understanding how such rapid changes impact biodiversity. Insects are recognized as serving important roles as indicators of overall ecological health. Specifically, the diversity and presence of ants can serve as indicators of environmental quality due to their ubiquitous presence across many terrestrial environments and their key role in numerous ecosystem processes. The aim of this study is to measure changes in ant biodiversity across an urban gradient in a temperate habitat. Food baits were set out across nine locations representing various degrees of urbanization as measured by percent impervious surface. Results show the abundance of ants significantly increased with increasing urbanization, with more generalist species found in great abundance in highly urbanized areas. However, a significant difference in ant species diversity across the urban gradient was not detected. The findings are consistent with other urban studies that showed an increase in the abundance of generalist species in urban areas. As the southeast United States continue to undergo urbanization to meet the demands of a growing human population, it is important for developers and ecologists to consider ways in which urban design can facilitate biodiversity, as levels of biodiversity will ultimately impact urban ecosystem function and the subsequent health of all occupants.