Walleye (Sander vitreus [Mitchill]) are native to rivers and streams in the Mobile River basin in Mississippi and Alabama. These populations comprise a genetically unique strain (Gulf coast walleye, GCW) and represent the southern-most distribution of walleye in the United States. Luxapallila Creek was considered an important spawning site for GCW prior to and shortly after impoundment of the Tombigbee River in 1980. Extensive sampling in Luxapallila Creek in 2001 and 2002 collected only one larval walleye. Microsatellite DNA analysis suggested 14 of 16 adult walleye from Luxapallila Creek were hatchery-produced fish or their progeny. Controlled angling catch rates of adult walleye have declined since 1997. The scarcity of wild-spawned walleye and the similarity of wild-caught and hatchery broodstock walleye indicates that the GCW population in, or spawning in, Luxapallila Creek is sustained by stocking and recruitment from these stocked fish may be diminishing.