The natural habitat as well as the food abundance and food sources of avian species is changing due to urbanization, and such anthropocentric actions could lead to devastating impacts on bird populations. As changes in distribution and nutrition are thought to be related to the gut microbiome, the goal of this study was to determine the relationship between nutritional markers, including body mass, gizzard mass, triglycerides, free glycerol and glycogen, and the gut microbiome in urban and rural house sparrows (Passer domesticus), to understand physiological differences between urban and rural house sparrows. We hypothesized that increased access to human refuse, through urbanization, may significantly alter the gut microbiome and thus, the nutritional physiology-the effects of foods on metabolism-of urban birds. Fecal samples were collected from rural (n=13) and urban (n=7) birds to characterize the gut microbiome and plasma samples were collected to measure nutritional markers using commercially available kits. Following euthanasia, liver samples were collected to measure triglycerides, free glycerol and glycogen. While there were no significant differences in circulating triglycerides or free glycerol between populations, urban birds had significantly greater blood glucose (p=0.046) compared to rural birds, when normalized to body mass. Additionally, rural birds had significantly more plasma uric acid (p=0.016) and liver free glycerol (p=0.044). Higher blood glucose suggests greater accessibility to carbohydrates in an urban setting or higher rates of gluconeogenesis. Uric acid is a byproduct of purine catabolism and a potent antioxidant. Thus, higher uric acid suggests that rural birds may utilize more protein for energy. Finally, higher liver free glycerol in rural birds suggests they metabolize more fat but could also indicate that urban birds have greater glycerol gluconeogenesis, which may consume free glycerol resulting in higher glucose concentrations. However, the current study does not provide evidence for this as there were no significant differences in the gluconeogenic enzyme PEPCK-C levels between urban and rural house sparrows (p= 0.165). While triglyceride, glucose, and uric acid levels differed between urban and rural birds, there were additionally no significant differences in the gut microbiome, indicating that although nutritional physiology can be affected by distribution and varying food availability and sources, differences in the gut microbiome are evident at the phyla level.