Blood antioxidant defenses and hematological adjustments in crowded/uncrowded rainbow trout (Oncorhynchus mykiss) fed on diets with different levels of antioxidant vitamins and HUFA.

Rainbow trout maintained at crowding or noncrowding conditions were fed on five experimental diets that were formulated considering two levels of vitamin E (25.6 and 275.6 mg/kg diet), vitamin C (0 and 1000 mg/kg diet) and HUFA (12.5 and 30.5 g/kg diet): -E-HUFA, -E+HUFA, +E-HUFA, +E+HUFA, -C+E+HUFA. Hematological parameters, the activity of some antioxidant enzymes and lipid peroxidation from RBC were evaluated. The SOD isoenzyme pattern was analyzed by nondenaturing PAGE. Hematological response to crowding stress was manifested by increased hemoglobin and RBC count in most of the crowded groups. Antioxidant enzyme activity was clearly affected by dietary HUFA levels, with uncrowded fish fed on +HUFA diets showing a higher SOD activity compared to those fed on -HUFA diets. In uncrowded groups, only one CuZn-SOD isozyme was detected, whereas in the crowded fish a great variability was revealed with up to five isozymes. G6PDH activity was increased in uncrowded -E+HUFA fish compared to the remaining groups. Lipid peroxidation was significantly increased in -E+HUFA fish regardless of fish density. Data supported the negative correlation of lipid peroxidation and hematocrit or hemoglobin explained by decreased erythrocyte stability. Dietary imbalances in vitamin E and HUFA supplementation may promote oxidative stress which triggers hematological deterioration, which in turn would affect the whole hematological status and ultimately fish welfare.