Double-crested cormorants (Phalacrocorax auritus) exhibit highly adaptive and opportunistic foraging behavior. This flexibility in foraging and increases in population size have led to conflicts with aquaculture and recreational and commercial fishing (Duffy 1995). Although double-crested cormorants roosting in the lower Mississippi Valley appear to have minimal negative impact on sport fisheries, they may have a significant impact on commercial aquaculture production in this region (Glahn and Brugger 1995, Glahn et al. 1998). In 2003, the U.S. Fish and Wildlife Service released the Final Environmental Impact Statement on doublecrested cormorant management allowing more flexibility in control of these birds in areas where they are negatively impacting aquaculture, habitat for nesting colonial waterbirds, and other public resources (U.S. Fish and Wildlife Service 2003). The U.S. Fish and Wildlife Service’s Final Rule expands the 1998 Public Resource Depredation Order (50 CFR 21.47) to permit control of double-crested cormorants at winter roost sites in the vicinity of aquaculture facilities. Populations of double-crested cormorants declined sharply in the 19th and early 20th centuries followed by several periods of population growth in the middle and later decades of the 20th century (Hatch 1995). Atlantic and Interior/Great Lakes migratory populations have seen the greatest increase in breeding pairs. Between the 1970s and 1990s, double-crested cormorant populations in the Atlantic increased 4-fold to more than 96,000 pairs (Hatch 1995). Although double-crested cormorants experienced a marked increase in population size from the 1970s to the 1990s, recent estimates suggest a reduction in the overall rate of growth (Tyson et al. 1999). Although most populations of double-crested cormorants are migratory, a resident population (P. auritus subsp. floridanus) estimated at 10,000–30,000 individuals occurs in Florida (Brugger 1995, Hatch 1995). Population estimates for Florida suggest stable resident populations of P. auritus subsp. floridanus with increasing numbers of wintering birds from migratory subpopulations (Brugger 1995). It is unclear whether or not the resident subspecies in Florida is a genetically distinct lineage, separate from migratory populations. If genetic differentiation is sufficient, it is possible that this resident population may warrant special consideration in management and any control efforts. In addition to the population in Florida, smaller colonies of nonmigratory birds have become established in other areas of the southeastern United States (e.g., Mississippi Delta, Reinhold et al. 1998; Louisiana, Hatch and Weseloh 1999). Determining genetic distinctiveness of diverse populations of double-crested cormorants and the extent of gene flow are important for regional management decisions (Hatch and Weseloh 1999). It is thought that individuals exhibit high fidelity to a colony site; although, no data exist to support this claim (Hatch and Weseloh 1999). Reduction of populations on breeding grounds might prove more feasible than reduction of wintering birds because double-crested cormorants nest in distinct colonies that can be readily accessed. Control efforts on breeding grounds would be most effective at reducing depredation if the natal areas of wintering birds can be identified. The number of band returns is insufficient to establish a relationship between double-crested cormorant nesting colonies in the northern United States and Canada and the wintering, depredating populations in the southeastern United States. Genetic markers have been used to associate wintering dunlin (Calidris alpina) and Canada geese (Branta canadensis) with breeding populations (Pierson et al. 2000, Wennerberg 2001). Sufficient genetic differentiation among breeding populations is necessary to correctly assign samples of wintering birds to their natal areas. An analysis of variation in mitochondrial DNA (mtDNA) found no evidence of genetic differences among migratory and nonmigratory populations (Waits et al. 2003). However, microsatellite loci are known to evolve rapidly and, thus, may reveal population structure even in the absence of mtDNA structure. For example, Goostrey et al. (1998) used highly polymorphic microsatellite markers to assess population structure and differentiation in European populations of great cormorants (P. carbo subsp. sinensis and P. carbo subsp. carbo). They detected high levels of variation both within and among populations suggesting the potential for detecting differences among populations of double-crested cormorants. Our primary objectives are to 1) characterize the genetic 1 E-mail: claygreen@txstate.edu 2 Present address: Department of Biology, Texas State University, San Marcos, TX 78666, USA 3 Present address: Department of Biology, University of Memphis, Memphis, TN 38152, USA