Because of high numbers of animals killed on Paynes Prairie State Preserve, Alachua County, Florida, the Florida Department of Transportation constructed a barrier wall-culvert system to reduce wildlife mortality yet allow for passage of some animals across the highway. During a one year study following construction, we counted only 158 animals, excluding hylid treefrogs, killed in the same area where 2411 road kills were recorded in the 12 months prior to the construction of the barrier wall-culvert system. Within the survey area lying directly in Paynes Prairie basin, mortality was reduced 65% if hylid treefrogs are included, and 93.5% with hylid treefrogs excluded. Sixty-four percent of the wildlife kills observed along the barrier wall-culvert system occurred at a maintenance road access point and along 300 m of type-A fence bordering private property. The 24 h kill rate during the post-construction survey was 4.9 compared with 13.5 during the pre-construction survey. We counted 1891 dead vertebrates within the entire area surveyed, including the ecotone between the surrounding uplands and prairie basin which did not include the barrier wall and culverts. Approximately 73% of the nonhylid road kills occurred in the 400 m section of road beyond the extent of the barrier wall-culvert system. We detected 51 vertebrate species, including 9 fish, using the 8 culverts after the construction of the barrier wall-culvert system, compared with 28 vertebrate species in the 4 existing culverts prior to construction. Capture success in culverts increased 10-fold from the pre-construction survey to the post-construction survey. Barrier wall trespass was facilitated by overhanging vegetation, maintenance road access, and by the use of the type-A fence. Additional problems resulted from siltation, water holes, and human access. These problems could be corrected using design modifications and by routine, periodic maintenance.