Sustainable development of any type that utilizes water or involves excavations in karst aquifer systems, such as the regional Floridan aquifer system (FAS), requires knowledge of preferential groundwater flow pathways that can extend adverse impacts beyond the development site and alter natural hydroperiods. Such pathways include fractures and other types of karst conduits that are associated with modern and relict sinkholes. Developments, including power plants and mines, that have not accounted for these features have caused induced recharge, altered hydroperiods and saltwater intrusion in the FAS, resulting in destruction of wetlands and adverse impacts to other surface waters, wildlife habitat and threatened and endangered species. This study analyzed indicators of preferential groundwater flow by considering surface expressions of underlying geological conditions (lineaments and modern sinkholes) in the FAS, which coincide with the United States southeastern coastal plain. Lineament mapping by Vernon (1951) and the Florida Department of Transportation (FDOT, 1973), incorporating analog mapping techniques and hardcopy prints of satellite imagery, preceded extensive urbanization, groundwater extractions, and mining in the region. All of these alterations limit the ability to identify fractures using lineaments by reducing groundwater discharges and vegetation indicative of those discharges. In this study, established methods for georectification, including control-point identification and spatial matching of scanned maps and remotely sensed images, were applied to these previously mapped lineaments. These results were applied to the environmentally sensitive karst study area of Citrus and Levy Counties, Florida in the southern extent of the FAS. Geospatial analyses of lineament distribution and modern sinkhole locations from the state database showed a dense network of lineaments with associated sinkholes throughout the study area and seven surrounding counties, including the proposed sites for a nuclear power plant and two mines in Levy County. Proposed excavations and water use for construction and operation of the power plant and mines would result in irreversible adverse environmental impacts on extensive depressional wetlands beyond the surface-footprint impact of these developments via these preferential flow pathways that were not evaluated during the review process.