Dietary copper requirements of juvenile grouper, Epinephelus malabaricus

To quantify dietary copper (Cu) requirements in grouper, Epinephelus malabaricus, copper sulfate was added to the basal diet at 0, 1, 2, 4, 6, 8, 10 and 20 mg Cu/kg diet providing the actual dietary value of 0.11, 1.66, 2.41, 4.37, 6.56, 8.97, 11.03 and 20.05 mg Cu/kg diet, respectively. Each diet was fed to triplicate groups of grouper (initial body weight 13.35 ± 0.01 g) in a recirculated rearing system for 8 weeks. The Cu concentration in rearing water was monitored during the feeding period and ranged from 1.0–1.5 μg/l. Weight gain (WG) and feed efficiency (FE) were higher (P < 0.05) in fish fed diets with 4.37 and 6.56 mg Cu/kg diet than those in fish fed diets with ≥ 11.03 and ≤ 1.66 mg Cu/kg diet. Hepatic thiobarbituric acid reactive substances (TBARS) value was lowest in fish fed diets with 4.37 and 6.56 mg Cu/kg diet, intermediate in fish fed diets with 11.03 and ≤ 1.66 mg Cu/kg diet, and highest in fish fed the diet with 20.05 mg Cu/kg diet. Differences between each of the three groups were statistically significant (P < 0.05). Both hepatic copper–zinc superoxide dismutase (Cu–Zn SOD) and plasma ceruloplasmin activities in fish fell into four groups. Cu–Zn SOD value was highest in fish fed the diet with 6.56 mg Cu/kg diet, second highest in fish fed the diet with 8.97 mg Cu/kg diet, intermediate in fish fed the diet with 20.05 mg Cu/kg diet, and lowest in fish fed the diet with 0.11 mg Cu/kg diet; plasma ceruloplasmin was highest in fish fed the diet with 6.56 mg Cu/kg diet, second highest in fish fed diets with 2.41, 4.37 and 8.97 mg Cu/kg diet, intermediate in fish fed diets with ≥ 11.03 mg Cu/kg diet, and lowest in fish fed diets with ≤ 1.66 mg Cu/kg diet. All values in each of the four groups were significantly different from the values of three other groups (P < 0.05). Analysis by broken-line regression of WG, hepatic SOD activity and TBARS value and linear regression of whole-body Cu retention of the fish indicate that the adequate dietary Cu concentration in growing grouper is about 4–6 mg Cu/kg diet.