Haplotype sequence differences.

The round goby, Neogobius melanostomus, is a successful invasive fish species. Originating from the Caspian and Black Sea, it is now distributed widely within European fresh- and brackish waters. The River Rhine was colonized in 2008 only a few years after the opening of the Rhine-Main-Danube canal and only four years after N. melanostomus was first reported in the upper Danube River. Its invasion history of the River Rhine was unclear because the species was first detected close to the Rhine river delta which would suggest a route of invasion other than via the Rhine-Main-Danube canal. To investigate the colonization history of N. melanostomus in the Rhine, we combined abundance estimates with molecular analysis. Abundance estimates found N. melanostomus to be dominant in the Lower Rhine (> 90% of all catches). Molecular analysis was done on 286 individuals from four different sites. Using the mitochondrial control region (d-loop), we found three different haplotypes in both Rhine sites. None of the potential invasive source populations in the rivers Danube and Trave exhibited all three haplotypes. The molecular data therefore supported a scenario of two different colonization directions. Our results show that the invasion history of the River Rhine is complex and warrants further investigation.