Aleutian mink disease (AMD) is a prominent infectious disease in mink farms. The AMD virus (AMDV) has been well characterized in Europe where American mink (Neovison vison) are an introduced species; however, in North America, where American mink are native and the disease is thought to have originated, the virus’ molecular epidemiology is unknown. As such, we characterized viral isolates from Ontario free-ranging mink of domestic, hybrid, and wild origin at two proteins: NS1, a nonstructural protein, and VP2, a capsid protein. AMDV DNA was detected in 25% of free-ranging mink (45 of 183), indicating prevalent active infection. Median-joining networks showed that Ontario AMDV isolates formed two subgroups in the NS1 region and three in the VP2 region, which were somewhat separate from, but closely related to, AMDVs circulating in domestic mink worldwide. Molecular analyses showed evidence of AMDV crossing from domestic to wild mink. Our results suggest that AMDV isolate grouping is linked to both wild endogenous reservoirs and the long-term global trade in domestic mink, and that AMD spills back and forth between domestic and wild mink. As such, biosecurity on mink farms is warranted to prevent transmission of the disease between mink farms and the wild.