The Great Plains of North America evolved with significant influence from bison (Bison bison), but is presently dominated by cattle (Bos taurus). While there are a variety of opinions concerning differences between these two species, there is a lack of scientific comparisons, including those that incorporate important ecological variation. We developed a framework to study and compare the grazing behavior and effects of bison and cattle within grassland ecosystems. Environmental (e.g., resource distribution, disturbance) and animal (e.g., number, social organization) factors play a critical role in determining grazing effects and should be incorporated into discussions that compare the effects of bison and cattle. Using this framework we specifically compare the grazing behavior of both species in tallgrass prairie and discuss the implications of these differences in the context of conservation. We collared bison and cattle with global positioning systems and used resource selection functions to estimate the importance of various environmental factors on site selection. Both species preferred recently burned areas and avoided steeper slopes. Cattle selected areas that were closer to water, while bison were not limited by distance to water; cattle also preferred areas with woody vegetation, while bison avoided them. Incorporating broad scale environmental complexity allows for an effective comparison of ecological differences between bison and cattle. While there are similarities and differences in these species, a comprehensive analysis of all conditions and scenarios is not possible. It is clear, however, that the greatest differences between these species will likely be evident from broad scale studies across complex landscapes. In addition to species, conservation and land managers need to consider other environmental factors that are critical to grazing effects and overall conservation.