| Seven trials were conducted to investigate the effects of different protein sources ( fish protein and soy protein) and some antinutritional factor such as soybean oligosaccharides (SBOS), soybean stachyose (SBS), soy raffinose (SR), soy saponin (SS), soybean isoflavone (SBIF), phytic acid (PA) on the expression of protein-digestive–related-enzyme genes (trypsinogen-1, chymotrypsinogen-1, elastase-1 precursor) and their enzyme activity in the liver and intestinal of Japanese flounder (Paralichthys olivaceus).meanwhile, some important correlation existed in antinutritional factors content in diets, growth performance, protein digestive performance and above these enzyme activity and gene expression were investigated. The results were summarized as follows:In trial 1, using FM, fish protein concentrate (FPC), SPC and soy protein isolate (SPI) as protein sources, four isonitrogenous(crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets (diets FM, FPC, SPC and SPI) were formulated to meet the protein and energy requirements of Japanese flounder. Hepatic elastase 1 precursor gene expression (HEPGE) in fish fed animal protein sources (APS)- based diets (diets FM and FPC) were significantly higher than those in fish fed plant protein sources (PPS)-based diets (diets SPC and SPI) (P<0.05). But hepatic trypsinogen-1 gene expression (HTGE) in fish fed APS- based diets lower than those in fish fed PPS-based diets (P<0.05). Hepatic trypsin activity (HTA), intestinal trypsin activity (ITA) and intestinal chymotrypsin activity (ICA) in fish fed APS- based diets were significantly higher than those in fish fed PPS-based diets (P<0.05).In trial 2, two isonitrogenous (crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets included only FM as sole protein source with (Diets FMO) or without (Diets FM) supplemental 10% SBOS were formulated. The results showed as follows: Hepatic chymotrypsinogen-1 gene expression (HCGE) was not significantly affected by supplementation with SBOS (P>0.05). SBOS supplementation did significantly affected HEPGE and HTGE in fish fed FM-based diets, significantly decreased HEPGE (P <0.05) and significantly increased HTGE in fish fed FMO diets (P<0.01). Supplementation with SBOS significantly decreased HTA (P< 0.01).In trial 3, adding 0.00%,0.5%, 1.0% and 1.5% SR to FM-based diet, thus four isonitrogenous(crude protein 42%) and isocaloric (gross energy 19.5 KJ/g) practical diets were formulated to meet the protein and energy requirements of Japanese flounder.Adding SR to FM-based diet did significantly affect digestive-relate enzyme of Japanese flounder (P<0.05), HTGE (r=0.924, P<0.01), HCGE (r=0.552, P<0.05), HEPGE (r=0.727, P<0.05) increased, and ITA (r=-0.970, P<0.05), ICA (r =–0.790, P<0.05), and HTA (r=-0.776, P<0.05) decreased with increasing dietary SR levels. There were the correlation existed between HTGE and Specific growth rate (SGR) (r=-0.761, P<0.05), Apparent digestibility coefficient of protein (ADCP) (r=-0.634, P<0.05); the significant correlation existed between HEPGE and SGR (r=-0.971, P<0.01); The significant correlation existed between HTA and PER (r=0.778, P<0.05) , the significant correlation also existed between ITA and ADCP (r=0.856, P<0.05), HTA (r=0.854, P<0.05) , ICA (r=0.667, P<0.05) , PER (r=0.772, P<0.01).In trial 4, adding 0.00 %, 0.4 %, 0.8 %, 1.5 % SBS to FM-based diet, thus four isonitrogenous(crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets were formulated to meet the protein and energy requirements of Japanese flounder. Adding SBS to FM-based diet did significantly affect digestive-relate enzyme of Japanese flounder, HTGE (r=0.617, P<0.05), HCGE smoothly increased, HTA (r=-0.931, P<0.05) and ICA (r=-0.998, P<0.01) decreased with increasing dietary SBS levels. There are the significant correlation existed between HTGE and ADCP (r=-0.683, P<0.05). Also there are the significant correlation existed between HEPGE and PER (r=-0.533, P>0.05), There are the significant correlation existed between ITA and ADCP (r=0.734, P<0.05), ICA (r=0.992, P<0.01). There were higher correlation between ICA and SGR(r=0.833, P<0.05), PER(r=0.934, P<0.05).In trial 5, Adding 0.0 %, 0.1 %, 0.3 %, 0.8 % SBIF to FM-based diet, thus four isonitrogenous(crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets were formulated to meet the protein and energy requirements of Japanese flounder.Adding SBIF to FM-based diet did significantly affect digestive-relate enzyme of Japanese flounder. HTGE (r=-0.732, P<0.05), HCGE and HEPGE (r=-0.666, P<0.05) smoothly decreased, HTA (r=0.778, P<0.05), ITA increased with increasing dietary SBIF levels. There are the significant correlation existed between HTGE and HTA (r=-0.968, P<0.01), HEPGE (r=0.861, P<0.05), HCGE (r=0.846, P<0.05), SGR(r=0.788, P<0.01), PER (r=0.778, P<0.05). The correlation coefficient existed between HTA and SGR (r=0.906, P<0.05), ADCP (r=0.734, P<0.05), PER (r=0.814, P<0.05). There were the significant correlation existed between HCGE and PER (r= -0.845, P<0.05), ADCP (r=-0.777, P<0.01). There were higher correlation between HEPGE and SGR (r=0.624, P<0.05), PER (r=0.856, P<0.05).In trial 6, Adding 0.0 %, 0.2 %, 0.4 %, 0.8 % phytic acid (PA) to FM-based diet, thus four isonitrogenous(crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets were formulated to meet the protein and energy requirements of Japanese flounder.Adding PA to FM-based diet did significantly affect digestive-relate enzyme of Japanese flounder. HTGE, HCGE, HEPGE (r=-0.850, P<0.05), HTA (r=-0.776, P<0.05), ITA (r=-0.914, P<0.01), ICA (r=-0.998, P<0.01) decreased, with increasing dietary PA levels. There were the significant correlation existed between HTA and ADCP (r=0.980, P<0.01), PER (r=0.836, P<0.01), SGR (r=0.947, P<0.05), NPU (r=0.837, P<0.05), synchronization, There were the significant correlation existed between ITA and PER (r=-0.772 P<0.05), FI (r=-0.987, P<0.01), ADCP (r=0.856 P<0.01), NPU (r= 0.987, P<0.01), ICA (r=0.992, P<0.01), SGR (r=0.750, P<0.05). There were correlation between ICA and SGR (r=0.738, P<0.05), ADCP (r=0. 755, P<0.05).In trial 7, Adding 0.0 %, 0.1%, 0.4 %, 0.8 % soy saponin (SS) to FM-based diet, thus four isonitrogenous (crude protein 49%) and isocaloric (gross energy 19KJ/g) practical diets were formulated to meet the protein and energy requirements of Japanese flounder. Adding SS to FM-based diet did significantly affect digestive-relate enzyme of Japanese flounder. HTGE (r=0.684, P<0.05), HCGE (r=0.882, P<0.05), HEPGE (r=0.829, P<0.05) increased, and HTA (r=-0.909, P<0.05), ITA (r=-0.991, P<0.01), ICA (r=-0.786, P<0.05) decreased with increasing dietary SS levels. There were the significant correlation existed between HTGE and ADCP (r=-0.767, P<0.05), PER (r=-0.734, P<0.05). The significant correlation also existed between HCGE and PER (r=-0.957, P<0.05), ADCP (r=-0.985, P<0.01), SGR (r=-0.632, P<0.05). There were the significant correlation existed between HEPGE and ADCP (r=-0.977, P<0.05), PER (r=-0.912, P<0.05). The significant correlation also existed between HTA and HCGE (r=-0.913, P<0.05), ADCP (r=0.969, P<0.05), SGR (r=0.692, P<0.05), PER (r=0.910, P<0.05). There are the significant correlation existed between ITA and PER (r=0.963, P<0.05), HTA (r=0.953, P<0.05), SGR (r= 0.854, P<0.05), ADCP (r= 0.944, P<0.05), ICA (r=0.719, P<0.05). There were the significant correlation existed between ICA and SGR (r=0.888, P<0.05), PER (r= 0.689, P<0.05),.
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