iTRAQ‐based identification of differentially expressed proteins related to growth in the swimming crab,Portunus trituberculatus

The swimming crab, Portunus trituberculatus, is an important farmed species in China, and it has been studied extensively, which requires more information on its genetic background. To date, information on the proteomics of P. trituberculatus is scarce. In this study, we used isobaric tags for relative and absolute quantitation (iTRAQ)‐based proteomics to investigate growth regulation in P. trituberculatus. Total proteins were isolated from five tissues (eyestalk, gill, heart, hepatopancreas and muscle). Equal quantities of protein from each tissue were pooled at the proteome level using the iTRAQ method. A total of 961 proteins were identified using liquid chromatography coupled with tandem mass spectrometry and de novo sequencing data. Using a 1.2‐fold change in expression as a physiologically significant benchmark, 30 differentially expressed proteins (DEPs) were found to undergo differential expression related to the growth of P. trituberculatus, and most of the growth‐related proteins, including those involved in metabolism, immune responses, DNA duplication and protein synthesis, were upregulated, indicating that conservation of energy is an important strategy to cope with growth. There was a high consistency between the expression levels determined using iTRAQ and mRNA, highlighting the high reproducibility of our proteomic approach and its great value in elucidating the molecular mechanisms of P. trituberculatus.     

Effects of low salinity stress on immune response and evaluating indicators of the swimming crab Portunus trituberculatus

The purpose of the present study was to investigate the effects of salinity stress on immune responses and evaluating indicators in swimming crab Portunus trituberculatus. The crabs (150 ± 8.5 g in body weight) were exposed to different salinities as 21, 26 and 31‰ (control) for 6 days. The results showed the total haemocyte counts (THC) and prophenoloxidase (proPO) activity in the haemocytes decreased significantly in the treatment groups after 6 hr, and reached the lowest levels at 12 hr. The phenoloxidase (PO) activity in the plasma increased significantly and peaked at 12 hr, then recovered to control level after 24 hr. The phagocytic per cent of haemocyte, antibacterial and bacteriolytic activities in the plasma decreased significantly in the treatment groups, and reached the lowest level at 12 hr, then recovered to control level after 72 hr. The dopamine (DA) and 5‐hydroxytryptamine (5‐HT) contents in the plasma increased significantly and peaked at 12 hr, then recovered to control level, while the noradrenaline (NE) content in the plasma had no significant change throughout the duration of the experiment. The DA and 5‐HT receptors were significantly up‐regulated in the treatment groups. The highest value of mRNA expression of DA and 5‐HT receptors occurred at 12 hr and recovered to control level after 24 hr. In addition, the cAMP and protein kinase A (PKA) contents in the haemocytes increased significantly and peaked at 12 hr, then recovered to control level after 72 hr. The phospholipase C (PLC) and protein kinase C (PKC) contents in the haemocytes increased significantly and peaked at 12 hr, then resumed to control level after 24 hr. These results speculated that biogenic amine (DA and 5‐HT) is likely to play an important role in immune modulation via cAMP/PKA signalling pathway or PLC/PKC signalling pathway when P. trituberculatus is exposed to low salinity and these results will provide scientific data for immune evaluation.     

Effect of dietary leucine on growth performance,hemolymph and hepatopancreas enzyme activities of swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to determine the dietary leucine requirement for juvenile swimming crabs reared in cement pools. Six isonitrogenous and isolipidic practical diets (430 g/kg crude protein and 70 g/kg crude lipid) were formulated to contain graded leucine levels which ranged from 16.7 to 26.7 g/kg (dry weight). Each diet was randomly assigned to triplicate groups of 60 juvenile swimming crabs (initial average weight 3.75 ± 0.12 g) that were stocked in rectangle plastic baskets. The results of the present study indicated that dietary leucine levels significantly influenced weight gain (WG) and specific growth ratio (SGR) (p < .05), crab fed the diet containing 22.7 g/kg leucine had significantly higher WG and SGR than those fed the other diets. Feed efficiency and protein efficiency ratio were not significantly affected by the dietary leucine levels (p > .05). Total protein, cholesterol, triglyceride and glucose in serum were significantly affected by the dietary leucine levels. Aspartate aminotransferase (AST) and alanine aminotransferase activities in hemolymph, AST and superoxide dismutase activities in hepatopancreas were significantly affected by dietary leucine levels; moreover, crab fed the 16.7 g/kg leucine diet had higher malondialdehyde in hemolymph and hepatopancreas than those fed the other diets. Crab fed the diet containing 24.9 g/kg leucine had higher phenoloxidase activity in hemolymph than those fed the other diets. Based on two‐slope broken‐line model of SGR against dietary leucine levels, the optimal dietary leucine requirement for growth was estimated to be 22.1 g/kg of the dry diet (corresponding to 51.4 g/kg of dietary protein on a dry weight basis). In summary, findings of this study indicated that dietary leucine could improve growth performance and antioxidant status.     

盐度对三疣梭子蟹生长、蜕壳及能量利用的影响

在实验室条件下,研究了盐度(15,20,25,30,35)对三疣梭子蟹(Portunus trituberculatus)幼蟹生长、蜕壳和能量利用的影响,探讨了三疣梭子蟹生长的最适盐度、蜕壳与生长的关系以及不同盐度水平下蟹的蜕壳同步性。实验时间为50 d,结果显示:1、盐度显著影响三疣梭子蟹的摄食、生长和能量利用。盐度20、25和30组,三疣梭子蟹的日摄食率(FId)较小,但其食物转化效率(FCEd)较高,其中盐度30组蟹的食物转化效率显著高于盐度15和35组(P<0.05),与盐度20和25组差异不显著(P>0.05)。盐度20、25和30组的能量吸收效率(K1)和净生长效率(K2)显著高于盐度15和35组。实验结束时,盐度30组三疣梭子蟹获得了较高的湿重、相对增重率和特定生长率,且显著高于盐度15和35组(P<0.05),而与盐度20和25组间的差异不明显(P<0.05)。回归分析表明,当水体盐度为26.3时,三疣梭子蟹特定生长率(SGRd)最大。2、不同盐度下,三疣梭子蟹幼蟹从Ⅶ期生长到Ⅹ期的蜕壳周期为18.9~23.5 d,各盐度组间的差异不明显(P>0.05),但盐度30组三疣梭子蟹蜕壳周期较其他盐度组缩短了3.1~4.6d。低盐(15)下,三疣梭子蟹出现MDS(蜕壳未遂)死亡,蜕壳受到抑制;而高盐(35)则抑制三疣梭子蟹新壳的硬化,盐度20和25组三疣梭子蟹的蜕壳同步性较好。实验结果初步表明,养殖生产中水体盐度控制在25左右有利于三疣梭子蟹的蜕壳同步及生长。     

Effects of dietary fish oil substitution with blending vegetable oils on growth performance,antioxidant enzyme activities and tissue fatty acid composition of juvenile swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to evaluate the effects of replacement of fish oil (FO) with blending vegetable oils (VOs) on growth performance, antioxidant enzyme activities and fatty acid composition in tissue of swimming crab Portunustrituberculatus. Five isonitrogenous and isolipidic diets were formulated to contain VOs (colza oil: palm oil: linseed oil = 4:2:1) to replace 0 (the control diet), 250, 500, 750 and 1000 g/kg of FO (defined D0, D25, D50, D75, D100). Three hundred juvenile swimming crabs (initial weight 2.34 ± 0.08 g) were randomly stocked and sorted into 300 individual rectangle plastic baskets in three cement pools. Each treatment has three replicates, one replicate has 20 swimming crabs, and each diet fed 60 crabs distributed in 60 baskets. The results indicated that crabs fed the control diet showed significantly higher survival, final body weight, per cent weight gain (PWG), specific growth rate and moulting frequency, crude protein and crude lipid contents in muscles than those fed the D75 and D100 VO diets (p < .05). Crabs fed the D25 VO diet showed significantly higher concentration of triglyceride, low‐density lipoprotein cholesterol and total protein, activities of superoxide dismutase, catalase and glutathione peroxidase (GSH‐Px) in haemolymph than those fed the control diet (p < .05). Fatty acid composition in hepatopancreas was positively correlated with dietary composition. In summary, based on the PWG, the optimal replacement of FO with VOs was estimated to be 250 g/kg. These findings demonstrated that swimming crabs make better use of FO than VOs.     

Effects of different dietary lipid sources on growth performance,antioxidant enzyme activities and biochemical composition of juvenile swimming crab,Portunus trituberculatus

An 8‐week feeding trial was conducted to evaluate the effects of dietary lipid sources on growth performance, antioxidant enzyme activities and biochemical composition of juvenile swimming crab Portunus trituberculatus of initial weight 2.34 ± 0.08 g. Four different diets were formulated to contain fish oil (FO), soybean oil (SO), linseed oil (LO) or palm oil (PO). The highest final body weight, weight gain, specific growth rate and molting frequency were observed in crabs fed the FO diet. Crabs fed the SO diet showed higher glutathione peroxidase, superoxide dismutase (SOD) and catalase (CAT) activities in both serum and hepatopancreas than those fed the FO diet. The lowest malondialdehyde concentration in hepatopancreas and serum were occurred at crabs fed the SO diet. Crabs fed the LO diet had significantly higher SOD and CAT activities in hepatopancreas compared with those fed the FO diet. Crabs fed the PO diet had the highest activities of fatty acid synthase and carnitine palmitoyltransferase 1 activities in hepatopancreas among all treatments. Fatty acid compositions both in hepatopancreas and muscle reflected those of diets. Overall, these findings demonstrated that physiological–biochemical characteristics and lipid metabolism were significantly regulated by different dietary lipid sources. Moreover, dietary SO and LO supplementation could improve antioxidant ability.     

Characterization of a γ‐aminobutyrate type A receptor‐associated protein gene,which is involved in the response of Portunus trituberculatus to CO2‐induced ocean acidification

The γ‐aminobutyrate type A receptor‐associated protein (GABARAP) is a ubiquitin‐like modifier implicated in membrane trafficking and fusion events involving the γ‐aminobutyrate type A receptor, autophagy and apoptosis. In this study, the gene encoding GABARAP was cloned from swimming crab Portunus trituberculatus (PtGABARAP) based on the expression sequence tag (EST). The full‐length cDNA of 664 bp includes a 5′ untranslated region (UTR) of 87 bp, a 3′ UTR of 223 bp with a poly(A) tail, and an open reading frame (ORF) of 354 bp encoding a polypeptide of 117 amino acids with a predicted molecular weight of 13.96 kDa. The deduced amino acid sequence shares high similarity (93%–100%) with GABARAPs from other species and includes a conserved Atg8 domain. In a phylogenetic analysis PtGABARAP clustered with GABARAPs from other species, and more widely with other GABARAP family proteins. The impact of elevated ocean acidification (OA) on P. trituberculatus behaviours was investigated, and real‐time RT‐PCR revealed that PtGABARAP expression was up‐regulated after OA exposure. Ocean acidification also caused crabs anxiety‐like behaviours, like the shoal average speed increase, preference for dark environment (scototaxis) and fast exploration. The results indicated that GABARAP might be involved in the interactions of GABA_A receptors and elevated‐CO₂ seawater.     

Effects of dietary fish oil replacement with blended vegetable oils on growth,lipid metabolism and antioxidant capacity of subadult swimming crab Portunus trituberculatus

A 120‐day trial was conducted to assess the effects of dietary fish oil replacement with vegetable oils on growth, lipid metabolism and antioxidant capacity of subadult swimming crab Portunus trituberculatus. Five isonitrogenous and isolipidic diets were formulated to replace 0, 250, 500, 750 and 1000g/kg of fish oil with a mixture of soybean and rapeseed oil (defined as D1–D5), and each treatment had 30 replicate crabs. Dietary fish oil replacement had no significant effects on growth of the crabs, while the D3 had the highest hepatosomatic index and total lipids in hepatopancreas. The triglyceride and lipase activities in hepatopancreas increased significantly with increasing dietary fish oil replacement. The D4 had the highest levels of alkaline phosphatase (ALP) and acid phosphatase (ACP) in the hepatopancreas, as well as the haemolymph ALP, ACP and peroxidase. The highest levels of haemolymph total antioxidant capacity, catalase and malondialdehyde were detected in D1. Total n‐3 polyunsaturated fatty acids levels in hepatopancreas decreased significantly with increasing dietary fish oil replacement. In conclusion, dietary fish oil replacement had no significant effects on growth of P. trituberculatus, and 500g/kg of fish oil replacement could improve antioxidant capacity, but excessive replacement level will enhance lipid accumulation and peroxidation in the hepatopancreas.     

Ovarian development and nutritional quality changes of the wild‐caught Portunus trituberculatus during the artificial fattening period

A female swimming crab (Portunus trituberculatus) with fully developed ovaries is considered one of the most nutritious types of seafood. The fishing yield of P. trituberculatus in China has always been high; however, its economic value has been low and unreliable due to poor ovarian development and low nutritional value. Artificial fattening is an effective way to improve the value of wild‐caught crabs, and this study focused on analysing the ovarian development and nutritional quality of the female P. trituberculatus during an artificial fattening period. Therefore, this study provides scientific evidence on the effects of artificial fattening on female P. trituberculatus and can guide the development of nutritional quality regulations for wild‐caught female crabs during the fattening process. The results showed that the ovaries of the wild female P. trituberculatus developed gradually. The ovarian morphology became bulkier, while the gonadosomatic index (GSI) and total edible yield (TEY) were significantly improved after artificial fattening. Significant changes in the nutritional components of the edible tissues were observed, and the moisture content in the ovaries and hepatopancreas decreased continually, while both the protein and fat content in the ovaries significantly increased. Moreover, the fat content in the hepatopancreas significantly increased. The saturated fatty acids(ΣSFA) in the ovaries and hepatopancreas had an increasing trend after fattening, while ΣPUFA and Σn‐3PUFA first decreased and then increased. In contrast, the ratio of Σn‐3PUFA to Σn‐6PUFA decreased steadily. However, the fatty acid composition of the muscles did not change significantly. The levels of all the amino acids increased continuously during the fattening process, and EAA increased significantly in the early stages of the fattening period. Consequently, all amino acids became nonlimiting amino acids after fattening. Furthermore, the ratio of EAA/NEAA in the ovaries increased continually until the end of the fattening period. On the other hand, the fatness and TEY increased significantly after artificial fattening. In addition, the nutritional value significantly improved with the exception of the high‐quality fatty acids in the ovaries and hepatopancreas, which were slightly lower in both.     

Identification of quantitative trait loci for growth‐related traits in the swimming crab Portunus trituberculatus

The swimming crab (Portunus trituberculatus) is an economically important species in Asian aquaculture. Implementing growth‐related traits of P. trituberculatus into genetic breeding programmes is an ongoing effort. We used a previously published genetic linkage map of P. trituberculatus, containing 55 simple sequence repeat (SSR) markers and 172 amplified fragment length polymorphism (AFLP) techniques to map quantitative trait loci (QTL) for growth‐related traits in a single full sibling F₂ family. Ten growth‐related traits were measured for QTL mapping. Composite interval mapping identified 9 QTL on the female map and 16 on the male map. Individual QTL with additive effects explained 11–38% of the phenotypic variance for various traits using the female parent's map, and from 1% to 21% using the male parent's map. Two QTL explaining a large percentage of variation in body weight were detected on chromosome 17 on the female map, and on chromosome 16 on the male map, and contributed 38% and 18% of the phenotypic variance respectively. This is the first study to report the detection and positioning of major QTL affecting growth in a true crab species (Brachyura). The mapping of growth‐related QTL in this study raises the possibility of improving the growth of P. trituberculatus through marker‐assisted selection.     

Effects of co‐supply ratios of swimming crab Portunus trituberculatus zoeae and Artemia on survival and growth of East Asian common octopus Octopus sinensis paralarvae under an upwelling culture system

Despite recent advances in culture technology for East Asian common octopus Octopus sinensis paralarvae using upwelling systems, securing suitable feed for the paralarvae is an unresolved issue. The zoea of the swimming crab Portunus trituberculatus is a good candidate for paralarval feed because of the high fecundity of the adult females. To investigate the effects of supplying P. trituberculatus zoeae and their feeding method on paralarvae, we cultured paralarvae with supplying different combination ratios of zoeae and Artemia (10:0, 7:3, 5:5, 3:7 and 0:10), and with or without supplementing rotifers using small‐scale (3‐L) upwelling systems. Paralarval survival rate and growth were improved when zoeae were supplied as the main feed, but reduced when the proportion of Artemia exceeded half the whole preys. Supplementing rotifers did not affect the paralarval survival and growth. Subsequently, paralarvae were cultured by supplying zoeae (partially augmented by Artemia) using three large (1‐kl) upwelling systems to assess their feeding effectiveness in juvenile octopus production. Paralarvae could be cultured at high survival rates of 77.1 ± 5.1% to reach benthic juveniles at 23 days after hatching. In conclusion, supplying P. trituberculatus zoeae augmented with Artemia under an upwelling culture system has great potential for juvenile octopus production.     

Effects of dietary DHA/EPA ratios on growth performance,survival and fatty acid composition of juvenile swimming crab (Portunus trituberculatus)

This study was conducted to investigate the effects of dietary docosahexaenoic to eicosapentaenoic acid (DHA/EPA) ratios on growth performance, survival and fatty acid composition of juvenile swimming crab (Portunus trituberculatus). Four isonitrogenous and isoenergetic experimental diets were formulated to contain different DHA/EPA ratios (0.70, 0.84, 1.06 and 1.25). There were three replicates (15 crabs per replicate) for each diet treatment. The crabs were fed (about 6–8% body weight) twice daily for 8 weeks. A good growth performance and feed utilization were observed in swimming crabs fed the diets with DHA/EPA ratios of 0.70 and 0.84. Crabs fed diet with 0.70 DHA/EPA ratio showed a significantly higher weight gain (WG) compared with the crabs fed the diet with DHA/EPA ratio of 1.25 (P < 0.05). The result of this study showed that the survival increased with decreasing the ratio of DHA/EPA. The DHA/EPA ratios in polar lipid from tissues were influenced by the dietary DHA/EPA ratios. Results of this study indicated that the growth performance and survival of juvenile swimming crab are correlated to the DHA/EPA ratio in the diets, and the best growth performance and survival were achieved with the ratios of 0.70–0.84.     

Effects of γ‐aminobutyric acid supplementation on the growth performance,serum biochemical indices and antioxidant status of pharaoh cuttlefish,Sepia pharaonis

This study evaluated the effects of dietary γ‐aminobutyric acid (GABA) on the growth performance, serum biochemical indices and antioxidant status of pharaoh cuttlefish, Sepia pharaonis. Cuttlefish were cultured in open‐culturing cement pool systems for 8 weeks. Six practical diets supplemented with graded levels of GABA (0, 20, 40, 60, 80 and 100 mg/kg) were formulated. Each diet was randomly assigned to triplicate groups of 60 cuttlefish (mean weight: 10.33 g), the cuttlefish were fed two times per day to apparent satiation. The results showed that the specific growth rate (SGR), weight gain (WG) and feed efficiency (FE) significantly increased with dietary GABA supplementation (p < .05). The survival rate (SR) and protein content in muscle significantly increased when 58.9 mg/kg GABA supplied. Moreover, the nitric oxide (NO) content and acid phosphatase (ACP) activity in serum were significantly increased with dietary GABA supplementation (p < .05), while the activity of aspartate aminotransferase (AST) in serum decreased significantly when supplied with GABA at 58.9 mg/kg (p < .05). In addition, dietary GABA improved antioxidation activity by significantly increasing the activities of superoxide dismutase (SOD) and catalase (CAT) but decreasing malondialdehyde (MDA) levels in the liver and gill (p < .05). On the basis of the quadratic regression analysis of FE, the optimum content of dietary GABA in S. pharaonis was estimated to be 55.3 mg/kg. The findings of this study demonstrated that dietary GABA had a positive effect on the growth performance, serum biochemical indices and antioxidant status of S. pharaonis.     

Influence of variation in water temperature on survival,growth and yield of Pacific white shrimp Litopenaeus vannamei in inland ponds for low‐salinity culture

There is considerable interest in the culture of whiteleg shrimp (Litopenaeus vannamei) in inland low‐salinity water in Alabama and other states in the Sunbelt region of the US. However, the growing season is truncated as compared with tropical or subtropical areas where this species is typically cultured, and temperature is thought to be a major factor influencing shrimp production in the US. This study, conducted at Greene Prairie Aquafarm located in west‐central Alabama, considered water temperature patterns on a shrimp farm in different ponds and different years; and sought possible effects of bottom water temperature in ponds on variation in shrimp survival, growth and production. Water temperature at 1.2 m depth in 22 ponds and air temperature were monitored at 1‐hr intervals during the 2012, 2013, 2014 and 2015 growing seasons. Records of stocking rates, survival rates and production were provided by the farm owner. Correlation analysis and linear mixed model analysis of variance were used. Results showed that hourly water temperatures differed among ponds. The range of water temperature in each pond explained 41% of the variance in average final weight of shrimp harvested from each pond. In conclusion, the results suggest that variation in water temperature patterns has considerable influence on shrimp growth and survival in ponds.     

Quantitative dietary leucine requirement of juvenile Pacific white shrimp,Litopenaeus vannamei (Boone) reared in low‐salinity water

The experiment was conducted to determine the leucine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low‐salinity water (0.50–1.20 g L^?1). Six diets were formulated to contain 410 g kg^?1 crude protein with fish meal, peanut meal and precoated crystalline amino acids with different concentration of l ‐leucine (16.72, 19.60, 22.06, 24.79, 27.28 and 30.16 g kg^?1 dry diet). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.38 ± 0.002 g), and the feed trial lasted for 8 weeks. The results indicated that the maximum weight gain was observed at 24.95 g kg^?1 dietary leucine group, whereas the diets containing higher leucine concentration conversely reduced the growth performance (P < 0.05). Moreover, the highest body protein content and body protein deposition and the lowest haemolymph AST and ALT activities were also found at 24.95 g kg^?1 dietary leucine group. With the increase in leucine in diets, a dose‐dependent increase was found in body lipid content and haemolymph urea concentration. The polynomial regression calculated using weight gain, feed efficiency and body protein deposition indicated that the optimal dietary leucine requirement for L. vannamei reared in low‐salinity water was 23.73 g kg^?1 leucine of dry diet, correspondingly 57.88 g kg^?1 of dietary protein.     

Biotic and abiotic regulation of a low‐density stream‐dwelling brown trout (Salmo trutta L.) population: effects on juvenile survival and growth

The effects of biotic (density‐dependent) and environmental (flow and temperature) factors on the apparent survival, mean length and size variation of a low‐density brown trout population in the juvenile stage, that is, from their first summer (0+) to the end of the second year (1+), were determined. Apparent survival was negatively related to the age class density during the three periods (first summer, first winter and second summer). A significant interaction between the mean flow and 0+ density highlighted a gradient towards strong density dependence acting on fish loss (i.e., mortality or migration) with decreasing summer flow. Conversely, no density dependence was reported at higher mean flows. The mean length was determined by density‐dependent and density‐independent (temperature and flow) factors throughout the study period. The negative relationship between fish length and intracohort density was highly significant during the three periods. The yearling (1+) density was negatively related to 0+ fish length measured after the first summer, suggesting intercohort effects. A positive effect of temperature on fish length was observed. Mean length after the summer seasons (0+ and 1+ fish) was also positively related to mean flow. Fish size variation around the mean measured with the coefficient of variation (CV) increased with increasing 0+ densities, both at the end of the first summer and the first winter. Results suggested that density‐dependent and density‐independent factors acted jointly on apparent survival and growth with a predominance of biotic processes. We discussed the potential implications of density‐dependent regulations on growth and survival for population resilience after catastrophic events.     

Dietary branched‐chain amino acid supplementation affects growth and hepatic insulin‐like growth factor gene expression in yellowtail,Seriola quinqueradiata

We investigated the effect of dietary supplementation with a branched‐chain amino acid (BCAA) mixture (valine, leucine and isoleucine) on juvenile yellowtail (Seriola quinqueradiata) growth and hepatic insulin‐like growth factor (IGF) gene expression. Total dietary BCAA content was 8.1, 8.5, 9.0 and 9.8 g   kg^?1 for the control, 0.4%, 1.0% and 2.0% BCAA experimental groups, respectively. Body weight was measured on day 77, after which fish were fasted for 3 days and refed; the livers were then removed 3 or 9 h later for analysis of hepatic gene expression. Body weight significantly increased in the BCAA‐supplemented groups. Hepatic expression of IGF1 and IGF2 significantly increased in the 2% BCAA group at 3 and 9 h after refeeding. In the 1% BCAA group, hepatic IGF1 expression tended to be higher at 9 h than in the control group after refeeding. Also, hepatic IGF2 significantly increased at only 9 h after refeeding in the 1% BCAA group. In conclusion, dietary supplementation with crystalline BCAAs increased growth and hepatic expression of IGF1 and IGF2. These results suggest that dietary crystalline BCAA supplementation would be a valuable addition to yellowtail aquaculture practices.