Underwater noise pollution is a growing problem in aquatic environments and as such may be a major source of stress for fish. In the present study, we addressed the effects of ship noise and continuous Gaussian noise on adrenal activity in three European freshwater species. Underwater ship noise recorded in the Danube River and two Austrian lakes was played back to fish at levels encountered in the field (153 dB re 1 μPa, 30 min). Post exposure cortisol secretion was compared with control situations. Cortisol was measured with enzyme immunoassay techniques (EIA, ng cortisol/l water/g fish) in extracted aquarium water with corrections for fish mass. In the first series, two hearing specialists, the common carp (Cyprinus carpio) and the gudgeon (Gobio gobio) and one hearing generalist, the European perch (Perca fluviatilis) were exposed to ship noise. The noise level was well above hearing thresholds in these species. In a second series, fish were exposed to continuous Gaussian noise at a similar level (156 dB) which is known to induce temporary hearing loss in hearing specialists. All three species responded with increased cortisol secretion when exposed to ship noise. Interestingly, no elevation was observed when fish were exposed to continuous Gaussian noise. Our results indicate that ship noise characterized by amplitude and frequency fluctuations, constitutes a potential stressor in contrast to continuous noise. Surprisingly, the data also demonstrate no apparent differences between species possessing excellent hearing abilities (hearing specialists) and species with poor hearing abilities like perch.