Red foxes were absent or rare in the southeastern United States until the late 1800s. Their origins potentially include natural population increase/expansion, translocations from Europe, and, eventually, 20th century fur farming. Previous studies have found no European haplotypes in North America, but few samples were sourced from the Atlantic coastal plain, closer to the source of putative introductions. Through analysis of mitochondrial DNA in 584 red foxes from this region, we identified indigenous haplotypes in ≥ 35% of foxes, 1 of 2 European haplotypes in 17% of foxes and fur farm haplotypes in ≥ 13% of foxes; another 35% of foxes had haplotypes potentially indigenous or native. In contrast, only 3 of 135 (2%) male foxes carried a single European Y chromosome haplotype. Most European and fur farm haplotypes were found near the densely human-populated coastal plain and Hudson River lowlands; most red foxes of the Appalachians and Piedmont had native eastern haplotypes. Our findings suggest that the more remote, upland populations primarily reflect indigenous red fox matrilines, whereas urban-associated populations in and around the mid-Atlantic coastal plain and Hudson lowlands reflect an admixture of native and nonnative maternal sources. Autosomal markers are needed to further elucidate the extent of European and fur farm introgression in the Appalachians and further west.