The ethology of Spinibarbus hollandi (S. hollandi), which is an important freshwater fish in South China, was investigated in a physical model that represents a 2.6 km-long reach of Jingshajiang River on a scale of 1:60. The study focused on the fish's preference to different substrate and flow conditions. Five types of substrate of different grain diameter (silver sand of 1 mm, rough sand of 3 mm small gravel of 40 mm, medium gravel of 50 mm, sand and cobble of 40 mm) were used in the experiments on substrate preference of young S. hollandiit. Flow velocity preference of young S. hollandiit and turbulence intensity effects were investigated as well. The total residency time of S. hollandi on each substrate is 0.4 h (rough sand), 0.8 h (medium gravel), 2.2 h (rough sand), 1 h (small gravel) and 0.3 h (silver sand), respectively. Preference of young S. hollandi to sand and cobble beds (P < 0.01) was found to be significant in the experiments. Consistent results on substrate selection were obtained both in experiments with colonies of fish and individual fishes. Regarding to flow preference, a flume with flat-bottom was used. It was found that young (2-year-old) individuals were in significant favor (P < 0.05) of a flow velocities in the range 0.3-0.6 m/s. In previous studies on flow preference of fish, the average velocity was used as the indicator of inductive velocity, favorite velocity and critical velocity. However, there is a growing consensus that turbulent flow fluctuations have important effects on flow preference. Moreover, natural flows are mostly characterized by turbulence. Therefore, the effects of turbulence on fish behaviour were also investigated in this study. Velocity fluctuations and turbulence intensity were used as indicators to further quantify S. hollandi's preference to flow conditions. It was found that S. hollandiit favored assembling in deepwater zones under static condition, but were more frequently present in zones with intensive turbulence under flow condition. Fishes were more frequently found in the curved conduit of the water tank which waters with high turbulence intensity but low average flow velocity. This suggests that flow fluctuations are more important than average flow velocity with respect to fish behaviour. Therefore, it is recommended to include turbulence in the description of fish response to complex flow conditions. The study results are of importance to the development of a fish habitat model for S. hollandi. Moreover, they indicate that heterogeneous river morphology and diverse flow conditions, natural or man-made, are effective measures to improve fish habitat.