Karst aquifers are important groundwater resources, but are vulnerable to contamination due to relatively rapid subsurface transport. Springs, points where the landscape and water table intersect and cold groundwater discharges, link aquifer systems with land surfaces and water bodies. As such, in many regions, they are critical to the viability of lakes, streams and cold-water fish communities. An understanding of where springs are located is important to watershed, fishery and environmental management efforts in karst regions. To better understand spatial distribution of springs and as a potential method for identifying variables that characterize locations of springs for improved land and watershed management, a nearest-neighbor analysis and a discriminant function analysis (DFA) of springs were conducted in Winona County, Minnesota, USA, a karst landscape. Nearest-neighbor analysis examined the spatial spring distribution. Twenty-two variables describing the locations of springs were analyzed to ascertain their ability to discriminate correct aquifer unit or bedrock unit classification for each spring. Springs were clumped with the highest densities in the lowest elevations. Springs were correctly assigned to aquifer units and bedrock units with eight and 11 landscape variables, respectively. Forest land cover was the only land cover type contributing to spring discrimination. Consideration of upland human activities, particularly in forested areas, on spring discharge along with a better understanding of characteristics describing spring locations could lead to better management activities that locate and protect springs and their important contributions to regional ecohydrology.