The genetic diversity of feral and ranch American mink was studied in order to detect gene flux among rivers, investigate the processes of invasion, and determine the possible effects of river barriers. Tissue samples of 78 feral American mink from 5 different river catchments and 18 ranch mink, collected between 2007 and 2011 in Biscay, northern Spain, were genotyped at 21 microsatellite loci. Lack of genetic differentiation of feral mink among the sites and high differentiation between feral and ranch mink was suggested. These results confirm that the mink population established on Butrón River at the beginning of the 1990s may be the origin of almost all the feral mink population within the study area. Additionally, the occurrence of American and European mink was used to analyse the effect of fragmentation on the population viability. The size and composition of the home range of male European mink was considered to model minimum viable units for presence/absence. Forty-two minimum viable units were randomly distributed among rivers in order to analyse the effect of fragmentation on mink occurrence. Barriers were mapped and classified as slight, moderate or absolute, depending on the effect on mink movement, and were introduced into the models. The presence of European and American mink depended on the non-fragmented main river stretches and the number of tributaries free from barriers. Results showed that fragmented rivers can be temporarily occupied but the likelihood of death means that these areas are only sink patches for mink.