Effects of dietary tryptophan levels on growth performance, serotonin metabolism, brain 5-HT and cannibalism activities in tiger puffer, Takifugu rubripes fingerlings

Abstract

Essential amino acid (EAA) requirements of aquaculture fish species can be used as a guideline for commercial feed formulation. Several EAA requirements have been quantified in tiger puffer (Takifugu rubripes). However, as the third limiting amino acid (AA) of many plant protein sources, tryptophan requirement of this species has yet to be quantified. This study was conducted to determine dietary tryptophan requirement of tiger puffer, and investigate the effect of dietary tryptophan supplementation on the cannibalism, brain serotonin (5-hydroxytryptpamine, 5-HT) metabolism and serum cortisol levels. Six experimental diets were prepared using fish meal, gelatin and corn protein concentrate as intact protein, and then supplemented with crystalline AA premix. The analyzed tryptophan levels in the experimental diets were 3.5, 4.1, 4.5, 5.5, 7.0 and 11.8 g/kg of dry diet. Experimental diets were randomly fed in triplicate to tiger puffer (initial average weight of 17.5 g) for 58 days. The weight gain (WG), feed conversion ratio (FCR) and protein efficiency ratio (PER) were significantly affected by dietary tryptophan, and the break points were observed at 5.07, 4.82 and 4.99 g/kg of dry diet, respectively. Compared to the basal diet, the cannibalism (the rate of bite mark appearance) was significantly lower and survival rate was significantly higher when fish were given diets containing 7.0 and 11.8 g/kg tryptophan. The concentrations of free tryptophan and phenylalanine in the brain exhibited a nonlinear increase in response to the incremental levels of dietary tryptophan. Brain 5-HT concentration was significantly affected by dietary tryptophan levels, but did not show a significant dose-dependent response. The expression of genes related to 5-HT metabolism (tph1a, ddc and aldh) and serotonin reuptake (sert) in the brain varied significantly and exhibited a nonlinear relationship with the graded levels of dietary tryptophan. In addition, serum cortisol levels showed a decreasing trend till the 5.5 g/kg tryptophan inclusion level, at which point there appeared to be the lowest level. In conclusion, based on nonlinear regression analysis of the WG, FCR and PER, dietary tryptophan requirement of tiger puffer was determined to be range between 4.82 and 5.07 g/kg of dry diet (10.48–11.03 g/kg of dietary protein). Dietary tryptophan supplementation above requirement could mitigate cannibalism and improve the survival of tiger puffer, which may be attributed to regulating the synthesis and catabolism of 5-HT and its availability in the synaptic cleft, and subsequently the reduction of serum cortisol content.