High mortality and impairment in growth rate during pikeperch (Sander lucioperca) ongrowing are among the major bottlenecks for its development in aquaculture. These failures may be related to high stress responsiveness since the rearing conditions are not yet optimized for this species. The objectives were to characterize the stress and immunological responses of pikeperch to major aquaculture modalities, and to identify the optimal aquaculture conditions for improving its welfare status. In a screening experiment, eight factors considered as relevant for the welfare of pikeperch were compared in two modalities using a fractional multifactorial design (28−4). Each experimental unit represented a combination of 8 factors in two modalities including grading, stocking density (15 vs 30 kg·m−3), feed type (sinking vs mid-floating), light intensity (10 vs 100 lux), light spectrum (red vs white), photoperiod (long vs short), dissolved oxygen (60 vs 90%) and temperature (21 vs 26 °C). Fish sampling occurred on days 36 and 63. Stress markers (glucose, cortisol and brain serotonergic activity), innate immune parameters (plasma lysozyme and complement activities) and expression of some immune genes were assessed. Light intensity and the type of feed clearly appeared as directive factors for pikeperch culture. A strong effect of the feed type was observed on growth parameters while survival was impacted by high light intensity. Light characteristics (intensity, spectrum and photoperiod) and temperature were identified as determining factors for physiological and immune markers. No obvious relation was established between stress status and growth parameters and further investigations are needed to improve management strategies of pikeperch culture and knowledge on the relations between environmental conditions, stress and immunity in percid fish.