Pikeperch broodstocks were exposed to different photoperiods: constant light (24L:0D), constant darkness (0L:24D), and 12 h light, 12 h darkness (12L:12D), for 40 days. Half of the broodstocks of each photoperiod were exposed to handling stress at a specific time of the day. Results showed that cortisol and lactate did not reveal any significant difference. However, glucose levels in females increased in the stress-free darkness period in comparison with stressful darkness photoperiods (0L:24D-s). Red blood cells in males and white blood cells in females showed a significant difference under different photoperiod regimes. Both sexes showed no significant difference in the differential count of leukocytes under different photoperiods and handling stress. Constant photoperiods and handling stress affected the hematological parameters, particularly, the number of lymphocytes and neutrophils in females. Our findings revealed that due to a long-term exposure to stressors, pikeperch brooders become adapted to stressful conditions.