Nearly 30% of emerging infectious disease events are caused by vector-borne pathogens with wildlife origins. Their transmission involves a complex interplay among pathogens, arthropod vectors, the environment and host species, and they pose a risk for public health, livestock and wildlife species. Examining habitat associations of vector species known to transmit infectious diseases, and quantifying spatio-temporal dynamics of mosquito vector communities is one aspect of the holistic One Health approach that is necessary to develop effective control measures. A survey was conducted from May to August, 2010 of the abundance and diversity of mosquito species occurring in the mixed-grass prairie habitat of the Smoky Hills of Kansas. This region is an important breeding ground for North Americas grassland nesting birds and, as such, it could represent an important habitat for the enzootic amplification cycle of avian malaria and infectious encephalitides, as well as spill-over events to humans and livestock. A total of 11 species, belonging to the three genera Aedes, Anopheles, and Culex, was collected during this study. Aedes nigromaculis, Ae. sollicitans, Ae. taeniorhynchus, Culex salinarius, and Cx. tarsalis accounted for 98% of the collected species. Multiple linear regression models suggested that mosquito abundances in the grasslands of the central Great Plains were explained by meteorological and environmental variables. Temporal dynamics in mosquito abundances were well supported by models that included maximum and minimum temperature indices (adjusted R²=0.73). Spatial dynamics of mosquito abundances were best explained by a model containing the following environmental variables (adjusted R²=0.37): ground curvature, topographic wetness index, distance to woodland, and distance to road. The mosquito species we detected are known vectors for infectious encephalitides, including West Nile virus. Understanding the microhabitat characteristics of these mosquito species in a grassland ecosystem will aid in the control and management of these disease vectors.