The impact of fish and cyclopoid copepod predation on zooplankton communities was evaluated using large-volume mesocosms (depth 9.5 m; volume 13 m3) in the Římov reservoir (Czech Republic). Two yearling roach and perch individuals introduced into mesocosms represented the fish treatment, which was compared to cyclopoid copepods (initial abundance of 2 ind.L−1) and a control with no initial addition of predators. Our results clearly support the hypothesis that planktivorous fish feeding leads to the suppression of large-bodied cladocerans. In the presence of fish, the cladoceran community changed from a dominance of large-bodied Daphnia spp. at the beginning to dominance by the smaller Bosmina longirostris at the end of the experiment. Chlorophyll-a concentration and rotifer abundances increased in the absence of daphnids. In the absence of fish, the presence of large-bodied cladocerans resulted in decreasing chlorophyll-a concentration. Although no significant differences were observed between cyclopoid abundances in treatments stocked with cyclopoids and the control, the proportion of large cladocerans clearly showed the effect of the manipulation. The similar trends in both of these treatments did not confirm the importance of cyclopoid predation in our experiment. The overall strong effect of fish over cyclopoid predation suggests the main role of fish predation in the forming of zooplankton communities and in turn impacting phytoplankton biomass in mesocosms.