Introduction of non-native species is an important cause of biodiversity decline in rivers. Separated by mountains and salt water, the freshwater fish fauna in Italy has experienced a natural isolation from fish in continental Europe. As a consequence, several Italian fish species have diverged from their European sister species, likely with unique adaptations to the local environment. Relatedly, the region is also susceptible to the invasion of non-native fish, and today almost half the fish species present are of non-native origin. Several of these non-native species have Italian congeners susceptible to competition and hybridizations, and, in the long run, displacements and extinctions. One such example is the Italian gudgeon (Gobio benacensis) and its European congener European gudgeon (Gobio gobio). During the last few decades the European gudgeon was introduced in Italian waters and has since spread rapidly, causing progressive declines in the Italian species. As for several other similar species pairs, little is known about potential differences in ecology and behavior. Here we study differences between Italian and European gudgeons in a controlled laboratory environment, using a combined open field and provoked escape response test, as well as tracking their sympatric survival over time in the hatchery. The smaller Italian gudgeon displayed a lower maximum swimming speed compared to the larger European gudgeon. The Italian gudgeon also experienced substantially higher sympatric mortality, resulting in only European gudgeon surviving to the end of the experiment. Contrary to expectations, no difference was seen in boldness and the Italian gudgeon displayed a higher movement activity, moving a larger distance in an open field test, compared to its European sister species. The reported differences could play a role in the replacement process, and may also have impacts on the surrounding ecosystem, and the prey and predators that have coevolved with the Italian species.