The aim of the present study was to investigate possible stressful effects on European sea bass Dicentrarchus labrax reared under constant darkness (0L-24D) and to examine the possible anti-stressful effect of dietary tryptophan (TRP) supplementation. Juvenile European sea bass (initial body weight 4.23 ± 0.032 g) were reared for 10 weeks under 0L-24D and 12L-12D and fed either a commercial diet (0.47% TRP) or the same diet supplemented with L-TRP (2.47% TRP). Results showed that lighting conditions had no significant effect on fish growth, while a depressive effect by the TRP supplementation was obvious. All fish populations reared under 0L-24D exhibited reduced body protein, lipid and ash content and increased food consumption. Reduced body lipids, food consumption and nutrient utilization were observed in TRP-supplemented fed fish, along with lower liver lipids. Dietary TRP enrichment significantly lowered liver saturated and monounsaturated acids and increased poly- and highly-unsaturated fatty acids, especially in combination with 0L-24D. These changes were also considerably reflected in carcass fatty acid composition.