Cities are rapidly expanding, and wildlife may experience different selection pressures in urban environments when compared to natural habitats. Phenotypic differences between urban and natural populations may occur because of the altered urban environment. Behavior, the activity of the hypothalamic-pituitary-adrenal (HPA) axis and body condition can be expected to differ between urban and natural habitats. We used the eastern chipmunk (Tamias striatus) to test for differences in behavior assayed from an open field test, hair and fecal cortisol concentrations, and body condition (size-corrected body mass), predicting that urban chipmunks would exhibit more exploratory behavior, higher cortisol concentrations, and higher body condition, than their counterparts from natural habitats. We sampled eastern chipmunks in 2 urban areas paired with natural habitats and subjected adult chipmunks to an open field test, collected hair and fecal samples for the determination of cortisol concentrations, and measured body size and body mass to estimate body condition. Eastern chipmunks in urban habitats had significantly different behavior, tending toward reduced locomotion and grooming, and greater latency, than their counterparts from natural habitats. Urban chipmunks also had lower fecal cortisol concentrations than those from natural habitats, and female chipmunks were in better body condition when captured in urban habitats. These results suggest that urban habitats may be relatively benign for urban chipmunks, perhaps because of reduced need for exploration and the availability of anthropogenic food subsidies associated with urban environments.