The development of hydrokinetic energy technologies (e.g., tidal turbines) has raised concern over the potential impacts of underwater sound produced by hydrokinetic turbines on fish species likely to encounter these turbines. To assess the potential for behavioral impacts, we exposed four species of fish to varying intensities of recorded hydrokinetic turbine sound in a semi-natural environment. Although we tested freshwater species (redhorse suckers [Moxostoma spp], freshwater drum [Aplondinotus grunniens], largemouth bass [Micropterus salmoides], and rainbow trout [Oncorhynchus mykiss]), these species are also representative of the hearing physiology and sensitivity of estuarine species that would be affected at tidal energy sites. We evaluated changes in fish position relative to different intensities of turbine sound as well as trends in location over time with linear mixed-effects and generalized additive mixed models. We also evaluated changes in the proportion of near-source detections relative to sound intensity and exposure time with generalized linear mixed models and generalized additive models. Models indicated that redhorse suckers may respond to sustained turbine sound by increasing distance from the sound source. Freshwater drum models suggested a mixed response to turbine sound, and largemouth bass and rainbow trout models did not indicate any likely responses to turbine sound. Findings highlight the importance for future research to utilize accurate localization systems, different species, validated sound transmission distances, and to consider different types of behavioral responses to different turbine designs and to the cumulative sound of arrays of multiple turbines.