Atlantic salmon and brown trout frequently co-occur in the same habitats and juveniles of both species are difficult to discriminate. Hybridization between the two taxa has also been widely documented especially in endangered populations hence the need for species-specific molecular markers. Here we show that the microsatellite marker SsAD486 has two species-specific alleles allowing the identification of Atlantic salmon, brown trout, and their hybrids throughout the European range of these species. Since this marker is polymorphic in Atlantic salmon populations from Western Atlantic, it could also help discriminating between North American and European salmon.