Recirculating aquaculture systems (RASs) are seen as a promising technology to address the societal and environmental challenges of aquaculture. However, this technology is mainly used in intensive monoculture and little knowledge is available in polyculture approaches. In this study, we studied survival, growth performance, and behaviour of juvenile pikeperch Sander lucioperca reared in RAS either in monoculture or in polyculture (associated with sterlet Acipenser ruthenus or tench Tinca tinca or both sterlet and tench). After 30 days, the survival rate was 100% for both monoculture and polyculture reared pikeperch. The mean final weights and the biomass gain of pikeperch were significantly lower for pikeperch alone (respectively 75.7 ± 2.7 g and 25.2%) or with sterlet (respectively 81.0 ± 3.2 g and 38.3%) than in combination with tench (respectively 85.7 ± 8.1 g and 50.1%) or with sterlet and tench (respectively 90.3 ± 16.4 g and 51.5%). Behavioural changes were also detected depending on the fish combination: there are fewer interactions between individuals of pikeperch when reared alone (0.34 ± 0.10) or in polyculture with sterlet (0.40 ± 0.02) in comparison to the other polyculture conditions (with tench [0.80 ± 0.20], or the two other species [0.70 ± 0.15]). The group structure also differed with a lower cohesion and homogeneity of the pikeperch group when they were reared in monoculture compared to polyculture modalities. No aggressive interaction was detected between pikeperch regardless of rearing modalities. In conclusion, due to positive effects on growth parameters and few behavioural changes in pikeperch, this study highlights that RAS polyculture is a relevant alternative production strategy for pikeperch compared to monoculture.