The diurnal distribution and diet of Hypseleotris spp. was examined over a 14-month period in Maroon Dam, a productive subtropical reservoir in south-east Queensland, Australia. Three distinct size classes of Hypseleotris were observed. The smallest (<16 mm standard length (SL)) fish undertake a vertical diurnal migration in the pelagic throughout the year, spending daylight hours near the surface and night hours in deeper waters. The diet of this size class consists almost exclusively of zooplankton (98.6% total prey volume), with cladocera and copepoda dominating the identifiable prey items. A larger (12–20 mm SL) size class group of subadult/adult fish occupies the near-shore littoral throughout the daylight hours but moves out of the sampled area during the night. The diet of this size class is more diverse (zooplankton 58.5%, macro-invertebrate 25.0%, other 16.5%). The largest size class (>20 mm SL) of adult Hypseleotris remains in the near-shore littoral throughout seasonal and diurnal cycles and has a more eclectic diet than the other two size classes (zooplankton 28.8%, macro-invertebrate 28.9%, other 42.3%). Spatial and temporal differences in the distribution and diet of these three size class groups are discussed with reference to the abundance and availability of suitable prey, intraspecific competitive exclusion, predator avoidance and water quality.