This study evaluated the dietary lysine requirement by measuring the plasma free lysine concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A basal diet containing 36.6 % crude protein (29.6 % crystalline amino acids mixture, 5 % casein and 2 % gelatin) was formulated to one of the seven L-amino acid based diets containing graded levels of lysine (0.72, 1.12, 1.52, 1.92, 2.32, 2.72 or 3.52 % dry diet). A total of 35 fish averaging 512 ± 6.8 g (mean ± SD) were randomly distributed into seven groups with five fish in each group. After 48 h of feed deprivation, each group of fish was fed one of the experimental diets by intubation at 1 % body weight. Blood samples were taken at 0, 5 and 24 h after intubation. Post-prandial plasma free lysine concentrations (PPlys, 5 h after intubation) of fish fed diets containing ≥ 2.32 % lysine were higher than those of fish fed diets containing ≤ 1.92 % lysine. Post-absorptive free lysine concentrations (PAlys, 24 h after intubation) of fish fed diets containing 2.32 and 3.52 % lysine were higher than those of fish fed diets containing ≤ 1.52 % lysine. The brokenline regression analysis on the basis of PPlys and PAlys indicated that the lysine requirement of rainbow trout could be 2.34 and 2.20 % in diet. Therefore, these results strongly suggested that the dietary lysine requirement based on the broken-line model analyses of PPlys and PAlys could be greater than 2.2 but less than 2.34 % (corresponding to be 6.01 % ≤, but ≤ 6.39 % in dietary protein basis, respectively) in rainbow trout. Also, these results shown that the quantitative estimation of lysine requirement by using PPlys and PAlys could be an acceptable method in fish.

Three experiments were conducted to determine the effects of dietary arginine concentrations on plasma free amino acid (PAA) concentrations in rainbow trout, Oncorhynchus mykiss (Walbaum). The first experiment was conducted to determine appropriate post-prandial and food deprivation sampling times in dorsal aorta cannulated rainbow trout averaging 519±9.5 g (mean±SD) at 16°C. Blood samples were taken at 0, 2, 3, 4, 5, 6 and 24 h after feeding (0 and 24 h blood samples were taken from the same group of fish). PAA concentrations increased by 2 h post-feeding and the concentration of all essential amino acids except histidine peaked at 5 h and returned to 0 time values by 24 h. In the second experiment dorsal aorta cannulated rainbow trout averaging 528±11.3 g (mean±SD) were divided into 6 groups of 4 fish to study the effect of dietary arginine levels on PAA. After 24 h food deprivation, each group of fish was fed one of six L-amino acid diets containing graded levels of arginine (0.48, 1.08, 1.38, 1.68, 1.98 or 2.58%) by intubation. Blood samples were taken at 0, 5 and 24 h after feeding. Post-prandial (5 h after feeding) plasma-free arginine concentrations (PParg) showed a breakpoint at 1.03% arginine in the diet and post-absorptive (24 h after feeding) plasma free-arginine concentrations (PAarg) showed a breakpoint at 1.38% arginine. PAarg increased linearly from fish fed diets containing arginine between 0.48% and 1.38%, and the concentrations remained constant from fish fed diets containing arginine at or above 1.38%, but were all below PParg at all time points. Results of the third experiment confirm the results that PParg concentrations from fish fed arginine deficient diets were higher than PAarg (0 or 24 h values). Thus, in contrast to mammals and birds, the PParg when arginine is present in the diet as the most limiting amino acid such that it severely limits growth, increases in plasma rather than decreases.

This study aimed to reevaluate the dietary methionine requirement by means of the plasma methionine and ammonia concentrations in surgically modified rainbow trout, Oncorhynchus mykiss. A total of 35 rainbow trout averaging 505 ± 6.5 g (initial body weight, mean ± SD) were randomly distributed into seven groups with five fish in each group. After 48 h of food deprivation, each group was fed one of seven L-amino acid-based diets containing graded levels of methionine (0.25, 0.40, 0.50, 0.60, 0.70, 0.80 or 0.95% of diet, dry matter bases) by intubation at 1% bodyweight. Blood samples were taken at 0, 5 and 24 h after intubation. Post-prandial plasma free methionine concentrations (PPmet, 5 h after intubation) and post-absorptive plasma free methionine concentrations (PAmet, 24 h after intubation) of fish fed diets containing 0.60% or more methionine were significantly (P < 0.05) higher than those of fish fed diets containing 0.50% or less methionine. PPmet and PAmet in fish fed diets containing 0.60% or higher methionine were not significantly different except the PPmet of fish fed a diet containing 0.95% methionine. Post-prandial plasma ammonia concentrations (PPA, 5 h after intubation) of fish fed diets containing 0.70% or more methionine were significantly higher than those of fish fed diets containing 0.60% or less methionine, and PPA of fish fed diets containing 0.25% and up to 0.60% methionine were not significantly different from each other. Broken-line model analyses on PPmet, PAmet, and PPA indicated that the dietary methionine requirement of rainbow trout was between 0.59% (1.69) and 0.67% (1.91) of diets (% dietary protein bases) when the diets contained 0.5% cystine.

This study was carried out to evaluate the dietary threonine requirement by measuring the plasma free threonine and ammonia concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A total of 70 fish (average initial weight 506±8.2 g) were randomly distributed into each of the 14 net cages (5 fish/cage). After 48 hours (h) of feed deprivation, each group was intubated at 1% body weight with one of the seven L-amino acid based diets containing graded levels of threonine (0.42%, 0.72%, 0.92%, 1.12%, 1.32%, 1.52%, or 1.82% of diet, dry matter basis). Blood samples were taken at 0, 5, and 24 h after intubation. Post-prandial plasma free threonine concentrations (PPthr) of fish 5 h after intubation with diets containing 1.32% or more threonine were significantly higher than those of fish intubated with diets containing 1.12% or less threonine (p<0.05). Post-absorptive free threonine concentrations (PAthr) after 24 h of intubation of the fish with diets containing 0.92% or more threonine were significantly higher than those of fish intubated with diets containing 0.72% or less threonine. Post-prandial plasma ammonia concentrations (PPA, 5 h after intubation) were not significantly different among fish intubated with diets containing 1.12% or less threonine, except the PPA of fish intubated with diet containing 0.42% threonine. Broken-line model analyses of PPthr, PAthr, and PPA indicated that the dietary threonine requirement of rainbow trout should be between 0.95% (2.71) and 1.07% (3.06) of diet (% of dietary protein on a dry matter basis).