1. Habitat fragmentation is a principal threat to biodiversity and artificial river barriers are a leading cause of the global decline in freshwater biota. Whilst the impact of barriers on diadromous fish is well established, impacts on river-resident fish communities remain unclear, especially for low-head barriers.

2. We examined the movement of five contrasting freshwater fish (topmouth gudgeon, European minnow, stone loach, bullhead, and brown trout) in an experimental cascade mesocosm with seven pools separated by small vertical barriers.

3. Passage rates differed significantly among species and increased with body size and sustained swimming speed (U_sus), ranging from an average of 0.2 passes/hr in topmouth gudgeon to 4 passes/hr in brown trout. A random-walk simulation indicated that barriers can result in net downstream movement and shifts in community composition.

4. Passage rates in brown trout were leptokurtic: i.e. most individuals were relatively sedentary while a small proportion showed frequent movements. Upstream passage rates of brown trout increased with body length and boldness, while fish with lower aerobic scope tended to move downstream. Passage rates showed significant individual repeatability in brown trout, independent of body size, indicating the potential for in-stream barriers to exert selective effects on fish populations.

5. Our results show that barrier effects can be more complex than simply blocking fish passage, and that river-resident fish can be impacted even by very small barriers. We show that fish passage depends on a wide range of morphological, physiological, and behavioural drivers, and that barriers can exert selective effects on these traits and cause shifts in community composition.

6. Policy implications. Barrier mitigation measures need to embrace inter- and intraspecific variation in fish passage to avoid inadvertent artificial selection on fish communities. Given the high abundance of low-head structures in river systems worldwide, a paradigm shift is needed to recognise the subtle impacts of small barriers on freshwater biodiversity. Removal of small barriers or nature-like fishways should allow better passage of the wider fish community compared to widely used salmonid-centric fish passage options.