Juvenile Swimming Crab Research Articles

The effects of water flow velocity and odor release frequency on the odor-tracking behavior of swimming crabs (Portunus trituberculatus)

Chemical-guided tracking is vital for critical activities such as foraging and mating in swimming crabs (Portunus trituberculatus), as it facilitates the detection and localization of odor sources. While water flow and odor release patterns are known to influence odor-tracking behavior, the specific mechanisms of their impact remain poorly understood. This study quantified the tracking behaviors of juvenile swimming crabs in response to varying water flow velocities (0.7 cm/s, 1.6 cm/s, and 2.5 cm/s) and odor release frequencies (0.5 Hz, 1.0 Hz, and 1.5 Hz). Results indicate that (1) at a flow velocity of 1.6 cm/s, crabs exhibited a higher proportion of tracking and success. Successful trackers at this flow velocity also demonstrated better tracking performance, including higher net-to-gross displacement ratios (NGDR), reduced gross displacements, shorter exit and track durations, and simpler tracking trajectories. (2) While odor release frequency did not significantly affect the proportion of tracking or success, the successful trackers at a higher frequency (1.5 Hz) had lower NGDR and higher gross displacements compared to those exposed to a lower frequency (0.5 Hz). (3) The successful trackers were not stastistically affected by the interaction between flow velocities and release frequencies. (4) In contrast to successful trackers, unsuccessful ones showed highly variable responses to flow velocity and odor release frequency, with generally inferior tracking performance. These findings provide valuable insights into the olfactory ecology of swimming crabs and can inform the optimized of aquaculture management and formulated feeds production.

Evaluation of cottonseed oil as a substitute for fish oil in the commercial diet for juvenile swimming crab (Portunus trituberculatus)

A six-week feeding trial was carried out to determine the feasibility of cottonseed oil (CSO) as a viable substitute for fish oil (FO) in the commercial diet of swimming crab. Ninety healthy swimming crabs (initial body weight 34.28 ± 0.59 g) were randomly assigned to 90 plastic baskets. Three isonitrogenous and isolipidic diets (450 g/kg protein and 120 g/kg lipid) were formulated replacing FO with CSO at 0%, 50% and 100% (CSO-0, CSO-50, and CSO-100), respectively. Each diet was randomly allocated to three replicates, each consisting of 10 crabs. Results indicated that crabs fed with CSO-100 diet had the lowest the percent weight gain (PWG), specific growth rate (SGR) and survival among all treatments (P < 0.05). Albumin (ALB), alkaline phosphatase (ALP), glucose (GLU), triglyceride (TAG), total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), non-esterified fatty acid (NEFA) contents and alanine amino transferase (ALT) activity in hemolymph were significantly affected by dietary substitution of FO with CSO (P < 0.05). The contents of total saturated fatty acids (SFA), total mono-unsaturated fatty acids (MUFA) and total long-chain polyunsaturated fatty acids (LC-PUFA) in the hepatopancreas and muscle were negatively correlated with the substitution level, whereas total n-6 polyunsaturated fatty acids (n-6 PUFA) and linoleic acid (18:2n-6) contents increased significantly with increasing levels of dietary substitution of FO with CSO (P < 0.05). Dietary substitution of FO with CSO resulted in changes in the composition of volatile substances in muscle, with 16 volatile substances in muscle significantly affected (P < 0.05). The relative expression of genes related to lipid synthesis such as fatty acid synthase (fas), acetyl-CoA carboxylase (acc) and glycerol-3-phosphate acyltransferase 1 (gpat1) in the hepatopancreas were significantly up-regulated in the CSO-50 group compared to other treatment groups (P < 0.05). The relative expression of fatty acid anabolism-related genes fatty acyl desaturase 2 (fads2) and elongase 4 (elovl4) were significantly down-regulated with the increase of dietary substitution of FO with CSO (P < 0.05). In conclusion, 50% substitution with CSO had no negative effects on growth performance, promoted lipid synthesis and metabolism, facilitated lipid accumulation. However, complete substitution of FO with CSO inhibited fatty acid synthesis and metabolism, resulting in a lower tissue LC-PUFA content and an altered composition of muscle volatiles.

Dietary Astaxanthin Can Promote the Growth and Motivate Lipid Metabolism by Improving Antioxidant Properties for Swimming Crab, Portunus trituberculatus.

This study aimed to assess the influence of varying dietary levels of astaxanthin (AST) on the growth, antioxidant capacity and lipid metabolism of juvenile swimming crabs. Six diets were formulated to contain different AST levels, and the analyzed concentration of AST in experimental diets were 0, 24.2, 45.8, 72.4, 94.2 and 195.0 mg kg-1, respectively. Juvenile swimming crabs (initial weight 8.20 ± 0.01 g) were fed these experimental diets for 56 days. The findings indicated that the color of the live crab shells and the cooked crab shells gradually became red with the increase of dietary AST levels. Dietary 24.2 mg kg-1 astaxanthin significantly improved the growth performance of swimming crab. the lowest activities of glutathione (GSH), total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and peroxidase (POD) were found in crabs fed without AST supplementation diet. Crabs fed diet without AST supplementation showed lower lipid content and the activity of fatty acid synthetase (FAS) in hepatopancreas than those fed diets with AST supplementation, however, lipid content in muscle and the activity of carnitine palmitoyl transferase (CPT) in hepatopancreas were not significantly affected by dietary AST levels. And it can be found in oil red O staining that dietary 24.2 and 45.8 mg kg-1 astaxanthin significantly promoted the lipid accumulation of hepatopancreas. Crabs fed diet with 195.0 mg kg-1 AST exhibited lower expression of ampk, foxo, pi3k, akt and nadph in hepatopancreas than those fed the other diets, however, the expression of genes related to antioxidant such as cMn-sod, gsh-px, cat, trx and gst in hepatopancreas significantly down-regulated with the increase of dietary AST levels. In conclusion, dietary 24.2 and 45.8 mg kg-1 astaxanthin significantly promoted the lipid accumulation of hepatopancreas and im-proved the antioxidant and immune capacity of hemolymph.

Evaluation of spray-dried blood meal for application in commercial-like feed for juvenile swimming crab (Portunus trituberculatus)

The objective of the present study was to evaluate the effects of replacing fish meal (FM) with spray-dried blood meal (SDBM) on growth performance, antioxidant capacity, digestive enzyme activity, tissue Fe content, gene expression related to inflammation, and the TOR pathway in juvenile swimming crab (Portunus trituberculatus). Four isonitrogenous (47.0% protein) and isolipidic (9.0% lipid) diets were formulated to contain different levels of SDBM, with FM replaced by SDBM at 0, 10%, 20%, and 30%, respectively. The results showed that crabs fed diet SDBM30 exhibited the lowest survival among all treatments, and crabs fed the SDBM20 diet had the highest final weight, percent weight gain, specific growth rate and feed efficiency among all treatments. In addition, the protein content of the crab hepatopancreas decreased significantly and the lipid content increased significantly as SDBM replaced FM in the diets. Compared to other diets, crabs fed the SDBM30 diet had the lowest activities of antioxidant enzymes and the highest concentration of malondialdehyde in the hepatopancreas and hemolymph. The Fe contents in hepatopancreas and gill increased significantly, and the color of the hepatopancreas got darker as replacement of FM with SDBM increased from 0% to 30%. The expression of genes related to TOR (tor, akt, s6, and s6k1) and antioxidant (gpx) pathways in the hepatopancreas were down-regulated significantly with increased replacement of dietary FM with SDBM. In contrast, expression levels of genes related to protein translation (4ebp1, eif4e1a, eif4e2, and eif4e3) and inflammation pathways (myd88, traf6, relish, and irak4) were up-regulated significantly in crabs fed diets with SDBM supplementation. In summary, replacing FM with less than 20% of SDBM can promote the growth, but can darken the hepatopancreas, negatively affect antioxidant capacity, and induce an inflammatory response.

Effects of dietary histidine level on growth, antioxidant capacity and TOR signaling pathway in juvenile swimming crabs, Portunus trituberculatus

An eight-week feeding trial was conducted to determine the optimal dietary histidine requirement of juvenile swimming crab (Portunus trituberculatus). Six isonitrogenous and isolipidic diets (45.0% crude protein and 8.0% crude lipid) were formulated to contain 0.43%, 0.68%, 0.97%, 1.24%, 1.45% and 1.79% histidine, respectively. Each diet was randomly assigned to triplicate groups of 30 juvenile crabs (approximately initial weight 17.63 ± 0.08 g). The results indicated that the highest percent weight gain (PWG) and specific growth rate (SGR) was presented in crabs fed the diet with 1.24% histidine (P < 0.05). Crabs fed the diet with 1.45% histidine showed remarkably higher activities of alanine transaminase (ALT) and aspartate transaminase (AST) in hemolymph than those fed the other diets (P < 0.05). Meanwhile, the concentrations of malondialdehyde (MDA) in hemolymph and hepatopancreas remarkably decreased with dietary histidine levels increasing from 0.43% to 1.79%, and activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC) and hydroxyl radical scavenging activity in hemolymph and hepatopancreas exhibited the significant trend of first increasing to 1.17% histidine and then decreasing (P < 0.05). The concentrations of carnosine significantly increased with the increase of dietary histidine levels (P < 0.05). Moreover, dietary histidine levels could significantly influence the mRNA levels of genes related to TOR pathway such as tor, akt, s6k1, s6, eif4e1a, eif4e2 and eif4e3, and antioxidant such as cat, cMnsod, gpx, trx and prx (P < 0.05). Two slope broken-line model analysis of PWG against the dietary histidine levels indicated that the optimal dietary histidine requirement was estimated to be 1.17% of the dry matter (2.60% of the dietary protein) for juvenile swimming crab. In conclusion, dietary 1.24% histidine can significantly increase the carnosine contents in muscle, hepatopancreas and hemolymph, improve antioxidant performance and activate the TOR signaling pathway.

Application prospect of replacement of fish meal with spray-dried egg meal in diets for swimming crab (Portunus trituberculatus)

An 8-week feeding trial was conducted to evaluate the effects of replacing fish meal with spray-dried egg meal (SEM) on growth performance, anti-oxidative capacity, mTOR pathway, and cholesterol metabolism in juvenile Portunus trituberculatus. The results showed that the highest percent weight gain was observed in crabs fed diet with 10% replacement of fish meal with SEM. Essential amino acids in the muscle and hepatopancreas significantly decreased with the increase of dietary replacement levels of fish meal with SEM. The lowest content of MDA in hepatopancreas was occurred at those fed diet with 40% replacement of fish meal with SEM, the activities of T-AOC, GSH-PX and content of GSH in hepatopancreas significantly increased with dietary SEM replacement fish meal increasing from 10% to 50%. The expression of akt, eif4e1a, eif4e2, eif4e3, and 4ebp1 significantly up-regulated, however, the expression of tor was showed significantly linear down-regulated, and the highest expression of akt, eif4e2 and 4ebp1 were found in crabs fed diet with 20% replacement of fish meal with SEM, the highest expression of eif4e1a was observed in those fed diet with 10% replacement of fish meal with SEM, the highest expression of eif4e3 was occurred in crabs fed diet with 30% replacement of fish meal with SEM. In cholesterol metabolism, the expression of abcg1 and lrp2 significantly down-regulated and the expression of npc1 significantly up-regulated with increase of replacement level of fish meal with SEM. The content of T-CHO in hemolymph and hepatopancreas significantly linear increased. In conclusion, based on quadratic broken-line regression analysis of PWG against the dietary replacement of fish meal with SEM, the optimal replacement level of fish meal with SEM was estimated to be 15.94% for juvenile swimming crab.

Effects of dietary isoleucine level on growth and expression of genes related to nutritional and physiological metabolism of swimming crab (Portunus trituberculatus)

A six-week feeding trial was conducted to evaluate the effects of dietary isoleucine level on growth, myofiber histology, and mRNA levels of genes related to amino acid, glucose and lipid metabolism in swimming crab (Portunus trituberculatus). Six isonitrogenous and isolipidic diets (45% protein and 8% lipid) were formulated to contain graded levels of isoleucine (Ile), with the analyzed levels of isoleucine being 0.69%, 1.06%, 1.32%, 1.78%, 2.03% and 2.38%, respectively. A total of 180 crabs (initial weight 16.30 ± 0.01 g) were housed individually in 180 plastic baskets with 30 crabs assigned to each diet, divided randomly into three replicates of 10 crabs. Each crab was fed twice at 08:00 and 18:00 with a daily feeding ration of 6–8% of crab body weight. Eight crabs from each replicate group were sampled and dissected after the feeding trial to provide hemolymph for biochemical analysis, hepatopancreas for proximate composition, amino acid profile enzyme activity, and gene expression, and muscle for proximate composition, amino acid profile and histology. The results indicated that final weight, percent weight gain and specific growth rate increased significantly as dietary isoleucine level increased from 0.69% to 1.78%, and then decreased with further increased dietary isoleucine. Crabs fed the diets with 1.78%, 2.03% and 2.38% isoleucine showed significantly higher muscle protein contents (17.80%, 17.99% and 17.65% of wet mass) than crabs fed lower levels of isoleucine. The highest concentrations of IGF1R and CHH in hemolymph (187.95 and 529.42 pg ml−1, respectively) and hepatopancreas (24.18 and 38.69 pg ml−1, respectively) were observed in crabs fed the diet with 1.32% isoleucine. Hepatopancreas expression levels of genes related to amino acid metabolism, including s6k1, bcat, alt and ast, were up-regulated in crabs fed 1.32% isoleucine compared to the other groups, whereas the expression level of tor decreased as dietary isoleucine increased. Expression levels of genes related to glucose intake, such as pi3k, igf1r and glut1, were significantly higher in crabs fed 1.32% isoleucine compared to crabs fed 0.69% isoleucine. Expression levels of genes related to gluconeogenesis (pepck and g6pase) were significantly higher in crabs fed 0.69% isoleucine than in crabs fed the diets with 1.32% and 1.78% isoleucine. In contrast, expression levels of genes related to glycolysis, including hk, gk and pk, were up-regulated in crabs fed 1.32% isoleucine compared to crabs fed 0.69% isoleucine. Gene expression levels of regulatory factors related to lipid metabolism (foxo, sirt1 and sirt3) were significantly higher in crabs fed 1.32% isoleucine compared to crabs fed the other treatments, while expression levels of genes related to lipolysis (hsl and cpt2) were significantly up-regulated in crabs fed the diets with 1.32% and 1.78% isoleucine. Expression levels of key lipogenesis genes (fas and acc) were down-regulated in crabs fed the diets with 1.32% and 1.78% isoleucine compared to those fed 0.69% isoleucine. Muscle histology indicated that crabs fed the diets with 1.32% and 1.78% isoleucine had significantly lower densities of myofibers (21.67 and 17.33 N/ mm2) compared to crabs fed 0.69% isoleucine. In conclusion, based on percent weight gain, the optimal isoleucine requirement of juvenile swimming crab was calculated to be 1.57% of diet (3.49% dietary protein) by quadratic curve model. Thus, appropriate dietary isoleucine levels (1.32–1.78%) promoted amino acid metabolism and protein synthesis, increased protein deposition, and altered myofiber structure of muscle and, in addition, promoted lipolysis and glycolysis, and reduced lipogenesis and gluconeogenesis in juvenile swimming crab.

Survival, Energy Status, and Cellular Stress Responses of the Juvenile Swimming Crab Portunus trituberculatus under Acute Nitrite Stress

Nitrite is a common pollutant encountered in aquaculture systems. During intensive hatchery, accumulation of nitrite can cause massive mortality of juvenile crustaceans. However, the nitrite toxicity and cellular stress responses in juvenile crustaceans is not clearly understood. Here, we investigate the survival, energy metabolism, and cellular stress responses in juvenile P. trituberculatus, an important aquaculture species in China, under acute nitrite stress. The results revealed nitrite resulted in a significant decrease in survival rate of juvenile swimming crab. After nitrite exposure, the activity of catabolic enzymes, such as HK, PK, CS, and CPT-1, were initially enhanced, and then they showed significant decrease at the late stage of exposure, accompanied by reduction in ATP and adenylate energy charge (AEC). The impaired energy homeostasis was possibly associated with disturbed AMPK signaling and enhanced anaerobic metabolism, which was indicated by the high levels of LDH activity and HIF-1α expression. Furthermore, we found that nitrite stress can depress antioxidant systems and unfold protein responses, causing oxidative damage and endoplasmic reticulum (ER) stress, and this, in turn, can trigger autophagy and apoptosis through both caspase-dependent and caspase-independent pathways. The results of the present study improve our understanding regarding adverse effects of nitrite on P. trituberculatus and provide valuable information for hatchery management.

Ammonia Stress Disturbs Moult Signaling in Juvenile Swimming Crab Portunus trituberculatus

Ammonia is a significant concern during hatchery culture in brachyuran species, and its accumulation may lead to abortive moulting and large-scale deaths of the early juveniles. To date, the underlying mechanism for ammonia-induced alteration of the moulting process is still unknown. In this study, we aimed to investigate the effects of ammonia on the moulting as well as the potential mechanisms in early juveniles of the swimming crab Portunus trituberculatus, an important aquaculture species in China. We evaluated the survival rate and moulting rate of the juvenile crabs (C2) and analyzed the expression pattern of the genes in key components of molt signaling during a complete moulting cycle under different concentrations of ammonia nitrogen (the control group: <0.1 mg/L; the LA group: 5 mg/L; and the HA group: 20 mg/L). The results showed that: (1) the survival rate in the LA and HA groups was lower than that in the control group at the end of the experiment, and moulting death syndrome (MDS) was only observed in the HA group; (2) the moulting rate was higher in the LA group and lower in the HA group compared to the control group; (3) consistent with the results of the moulting experiment, MIH showed decreased expression, and genes related to ecdysteroid synthesis, ecdysteroid receptors, and responsive effectors exhibited increased expression in the LA group compared to the control group; and (4) although MIH expression was upregulated, increased expression of the genes associated with ecdysteroid synthesis, ecdysteroid receptors and downstream effectors still observed in the HA group. Our results indicated that low levels of ammonia can promote moulting in juvenile swimming crabs by inhibiting the expression of MIH and activating moult signaling, whereas high levels of ammonia inhibit moulting and lead to MDS through impairing moult signaling.

Application evaluation of linseed oil as a substitute for fish oil in the commercial diet for juvenile Portunus trituberculatus

Vegetable oil (VO) is increasingly replacing fish oil (FO) in the commercial diet of crustaceans, partly due to a global shortage of FO. But little is known about how different amounts of substitution affect the health and metabolism of crustaceans. A six-week feeding trial was conducted to evaluated the effects of different replacement levels of FO with linseed oil (LO) on growth performance, antioxidant and digestive capacity and fatty acid metabolism in swimming crab, Portunus trituberculatus. Five isonitrogenous and isolipidic diets (480 g kg−1 protein and 120 g kg−1 lipid) were formulated to contain graded levels of LO, and FO was replaced with LO at 0%, 25%, 50%, 75% and 100%, respectively. The results showed that 50% of FO could be replaced by LO without affecting the growth performance and feed utilization for juvenile swimming crab. In hemolymph, 100% dietary replacement of FO with LO, the activities of alanine aminotransferase (ALT) and aspartate transaminase (AST) exhibited significantly higher than those fed the other diets, the concentrations of triacylglycerol (TAG), total cholesterol (T-CHO) and high-density lipoprotein cholesterol (HDL-C) significantly increased with increase of dietary replacement of FO with LO, which may lead to hyperlipidemia in swimming crabs. The number of R cells and their lipid droplets in hepatopancreas gradually increased with increase of replacement of FO with LO, the results confirmed the occurrence of lipid accumulation in the hepatopancreas. The activities of glutathione (GSH), and glutathione peroxidase (GPX) in hepatopancreas significantly decreased with increase of replacement of FO with LO, and crabs fed diet with 50% replacement of FO with LO had higher activities of trypsin and lipase (LPS) than those fed the other diets. Long-chain polyunsaturated fatty acids (LC-PUFA), n-3/n-6 PUFA in hepatopancreas and muscle significantly decreased with the increase of substitution level of FO with LO. Moreover, the expression of genes related to LC-PUFA synthesis as well as fatty acid β-oxidation, such as elongase 4 (elovl4), hepatocyte nuclear factor 4-α (hnf4α), retinoid X receptor (rxr), carnitine palmitoyltransferase 2 (cpt2) and acyl-CoA oxidase (acox2), were down-regulated with increase of dietary replacement of FO with LO. In conclusion, it is suggested that the optimal substitution ratio of LO for FO should be less than 50% in the diet of Portunus trituberculatus.

Evaluation of Krill Meal in Commercial Diets for Juvenile Swimming Crab (Portunus trituberculatus).

An 8-week feeding trial was carried out to assess the effect of dietary krill meal on growth performance and expression of genes related to TOR pathway and antioxidation of swimming crab (Portunus trituberculatus). Four experimental diets (45% crude protein and 9% crude lipid) were formulated to obtain different replacements of fish meal (FM) with krill meal (KM); FM was replaced with KM at 0% (KM0), 10% (KM10), 20% (KM20), and 30% (KM30); fluorine concentration in diets were analyzed to be 27.16, 94.06, 153.81, and 265.30 mg kg-1, respectively. Each diet was randomly divided into 3 replicates; ten swimming crabs were stocked in each replicate (initial weight, 5.62 ± 0.19 g). The results indicated that crabs fed with the KM10 diet had the highest final weight, percent weight gain (PWG), and specific growth rate (SGR) among all treatments (P < 0.05). Crabs fed with the KM0 diet had the lowest activities of total antioxidant capacity (T-AOC), total superoxide dismutase (SOD), glutathione (GSH), and hydroxyl radical scavenging activity and had the highest concentration of malondialdehyde (MDA) in the hemolymph and the hepatopancreas (P < 0.05). In the hepatopancreas, the highest content of 20:5n-3 (EPA) and the lowest content of 22:6n-3 (DHA) were shown in crabs fed with the KM30 diet among all treatments (P < 0.05). With the substitution level of FM with KM gradually increasing from 0% to 30%, the color of the hepatopancreas changed from pale white to red. Expression of tor, akt, s6k1, and s6 in the hepatopancreas was significantly upregulated, while 4e-bp1, eif4e1a, eif4e2, and eif4e3 were downregulated with dietary replacement of FM with KM increasing from 0% to 30% (P < 0.05). Crabs fed with the KM20 diet had notably higher expression of cat, gpx, cMnsod, and prx than those fed with the KM0 diet (P < 0.05). Results demonstrated that 10% replacement of FM with KM can promote growth performance and antioxidant capacity and notably upregulate the mRNA levels of genes related to TOR pathway and antioxidant of swimming crab.

Dietary phenylalanine level could improve growth performance, glucose metabolism and insulin and mTOR signaling pathways of juvenile swimming crabs, Portunus trituberculatus

An 8-week feeding trial was conducted to determine the optimal dietary phenylalanine requirement of juvenile swimming crab (Portunus trituberculatus). Six experimental diets (45.0% crude protein and 8.0% crude lipid) were formulated to contain 0.89%, 1.15%, 1.41%, 1.64%, 1.90% and 2.18% phenylalanine, respectively. Each diet was randomly divided into triplicate groups with 30 juvenile swimming crabs (initial weight 22.87 ± 0.03 g). The highest percent weight gain (PWG) and feed efficiency (FE) were presented in crabs fed with 1.64% phenylalanine diet, and the lowest PWG and FE were observed in crabs fed diet with 0.89% phenylalanine (P < 0.05), while no significant difference was presented in survival among all treatments (P > 0.05). Crabs fed the diet containing 0.89% phenylalanine had the lowest content of crude lipid in hepatopancreas among all treatments (P < 0.05). Hematological parameters related to glucose and lipids metabolism and enzyme activities involved in glycolysis and gluconeogenesis were significantly affected by dietary phenylalanine levels (P < 0.05). The contents of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), n-6 polyunsaturated fatty acids (n-6 PUFAs) and n-3 polyunsaturated fatty acids (n-3 PUFAs) in the hepatopancreas were notably affected by the dietary phenylalanine levels (P < 0.05). The enzyme activities related to glycolysis, gluconeogenesis and glycogen content in hepatopancreas were significantly influenced by dietary phenylalanine levels (P < 0.05). The mRNA levels of genes related to glycolysis and gluconeogenesis in the hepatopancreas were significantly affected by dietary phenylalanine levels (P < 0.05). Moreover, the insulin and mammalian target of rapamycin (mTOR) signaling pathway were notably activated by dietary phenylalanine levels (P < 0.05). Based on two slope broken-line regression analysis of PWG against the dietary phenylalanine levels, the optimal dietary phenylalanine requirement was estimated to be 1.60% dry matter (3.55% dietary protein) for juvenile swimming crab.

Interactive effects of dietary cholesterol and phospholipids on growth and metabolism of juvenile swimming crab, Portunus trituberculatus

Cholesterol and phospholipid play an important role in growth, molt and reproduction for crustaceans, although dietary optimum requirement of cholesterol and phospholipids has been determined for Portunus trituberculatus, but it is still unclear whether there was an interaction on each other. Therefore, an eight-week feeding trial was conducted to evaluate the interactive effects of dietary cholesterol and phospholipid levels on growth performance, fatty acid composition in hepatopancreas and the expression of genes related to cholesterol and phospholipid metabolism of Portunus trituberculatus. Six isonitrogenous and isolipid diets were formulated to contain two cholesterol levels (0 % and 0.5 %) and three phospholipid levels (0 %, 2 % and 4 %). The results indicated that specific growth rate (SGR) significantly increased with the increase of dietary phospholipid levels, however, there were no interactive effects between dietary cholesterol and phospholipid on percent weight gain (PWG) and SGR (P > 0.05). The concentration of total-cholesterol (T-CHO) and triacylglycerol (TAG) in hemolymph significantly affected by the interaction between cholesterol and phospholipid (P < 0.05), crabs fed diets with 0.5 % cholesterol supplementation had higher low-density lipoprotein cholesterol (LDL-C) concentration in hemolymph than those fed diets without cholesterol supplementation regardless of phospholipid supplementation (P < 0.05). TAG and LDL-C concentration in hepatopancreas were significantly affected by the interaction between cholesterol and phospholipid (P < 0.05), crabs fed diets with 0.5 % cholesterol supplementation had significantly higher T-CHO and LDL-C concentration in hepatopancreas than those fed diets without cholesterol supplementation (P < 0.05). There was a significant interaction between dietary cholesterol and phospholipid on activity of catalase (CAT) and the concentration of glutathione (GSH) and malondialdehyde (MDA) in hepatopancreas (P < 0.05), and crabs fed diets with cholesterol supplementation had higher superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in hepatopancreas than those fed diets without cholesterol supplementation (P < 0.05). Moreover, EPA, LC-PUFA, n-3 PUFA, n-3/n-6 PUFA and DHA/EPA in hepatopancreas were significantly affected by the interaction between dietary cholesterol and phospholipid (P < 0.05). The expression of adenosine triphosphate binding cassette transporter G1 (abcg1), C-type 1 class Niemann-Pick protein (npc1), scavenger receptor B (srb) and low-density lipoprotein receptor (ldlr) were up-regulated by dietary cholesterol and phospholipid levels (P < 0.05). In conclusion, the results indicated that dietary appropriate cholesterol and phospholipid levels could significantly improve growth performance and promote cholesterol metabolism.

New Insight Into the Molting and Growth in Crustaceans: Regulation of Energy Homeostasis Through the Lipid Nutrition

Organisms can regulate mitochondrial metabolic adaptation, further ameliorate the energy homeostasis to control the ATP production for the energy expenditure during the growth process under different physiological and nutritional conditions. Based on this understanding, the objective of the present study was to investigate how different dietary n-3 PUFA (polyunsaturated fatty acid) lipid sources modify the mitochondrial metabolic adaptation, and further affect the energy homeostasis and the growth of swimming crab (Portunus trituberculatus). A total of 120 swimming crab juveniles with an average initial weight of 10.17 ± 0.12 g were fed one of three diets (4 replicates/treatment) containing either fish oil (control), krill oil or linseed oil as dietary lipid sources for 8 weeks, and the effects of dietary lipid sources on the growth and energy homeostasis via the regulation of mitochondrial metabolic adaptation were evaluated. The study revealed that, compared with linseed oil rich in 18:3n-3, fish oil and krill oil rich in 20:5n-3 and 22:6n-3 significantly promoted the molting and growth of juvenile swimming crab, increased the ATP level, mitochondrial membrane potential, NAD+ substrate level, NAD+/NADH ratio and the mitochondrial DNA copy number. Furthermore, crabs fed the diet supplemented with krill oil can up-regulate the expression levels of genes related to energy metabolism. In addition, dietary krill oil also specifically improved the ability for scavenging free radicals produced in the process of physiological metabolism, reduced the level of lipid peroxidation and the degree of DNA oxidative damage, and improved the health status of swimming crab. The present study revealed the adaptation of mitochondrial metabolism and the regulation of the energy homeostasis of swimming crab to different dietary n-3 PUFA lipid sources, and provided a new insight into the relationship between the growth as well as molting and the energy homeostasis, which provided a novel insight into the lipid nutrition and energy metabolism of crustacean species.

Investigation of the Light Intensity Effect on Growth, Molting, Hemolymph Lipid, and Antioxidant Capacity of Juvenile Swimming Crab Portunus trituberculatus

An eight-week experiment was conducted to evaluate the effects of eight light intensities (0, 0.83, 1.61, 8.17, 15.89, 31.38, 63.69, and 124.31 μmol m-2 s-1) on the survival, growth, antioxidant, molting-related and apoptosis-related gene expression of juvenile swimming crab Portunus trituberculatus. The results showed that the survival rate of crabs was the highest under the light intensities of 8.17 and 15.89 μmol m-2 s-1. Under this light intensity, the crabs had the lowest hemolymph glucose (GLU) but the highest triglyceride (TG) content. The crabs in these treatments also had the highest total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT) activities, and the lowest malondialdehyde (MDA) content. The growth performance (including weight gain and specific growth rate) and molting frequency of crabs increased with increasing light intensity. The improved growth performance and molting of 8.17 and 15.89 μmol m-2 s-1 groups were also accompanied by up-regulated of the retinoid-X receptor (rxr), ecdysone receptor (ecr), nuclear receptor E75 (e75) gene expression, and down-regulated molt-inhibiting hormone (mih) gene expression. In addition, suboptimal light intensity (0, 0.83, 63.69, and 124.31 μmol m-2 s-1) significantly up-regulated the expression of apoptosis-related genes including c-Jun N-terminal kinase (jnk), tumor suppressor p53 (p53), and B-cell lymphoma-2 (bcl-2). Taken together, the suitable light intensity for the juvenile P. trituberculatus was estimated to be 9.96 μmol m-2 s-1.

An evaluation of the preferences of the juvenile swimming crab, Portunus trituberculatus, for different structural properties of shelters

Crustaceans select shelters non-randomly, and their preferences may be mediated by the structural properties of shelters. Providing shelters is an effective approach to improving the survival of pond-raised crabs, but the structural properties of shelters that would significantly benefit the survival rate remain unclear. Here, a laboratory experiment was conducted to examine the preference of swimming crabs (Portunus trituberculatus) for three types of shelter with different depths, heights, and widths using an animal behavioral observation system. The residence time of crabs in each shelter was calculated, the changes in their preference were described by the program PFunc, and the correlations between the carapace width and preference function traits were analyzed. The results showed that juvenile swimming crabs (carapace width = 5.08 ± 0.34 cm) preferred shallow, high, and wide shelters, and the strongest preferences observed were for depth and height. There was a correlation between crab carapace width and preference function traits, which suggests that preferences became stronger as the carapace width increased. Height and width are the two major dimensions that need to be controlled when designing a shelter, and their value should be determined by the carapace width of the crabs. Providing the crabs with shelter at their preferred dimensions in the pond can prolong the residence time, which may help to reduce the frequency of agonistic encounters and improve the survival of the crabs.

Effects of Dietary Carbohydrate Levels on the Growth and Glucose Metabolism of Juvenile Swimming Crab, Portunus trituberculatus

This experiment was conducted to evaluate the effects of different carbohydrate levels on the growth performance, glycogen accumulation in tissue, enzyme activities, and expression levels of genes related to hepatopancreatic glucose and lipid metabolism and insulin signaling pathway in swimming crab, Portunus trituberculatus for 8 weeks. The results indicated that crabs fed with the 60.0 g kg-1 carbohydrate diet exhibited the highest final body weight (FBW), percent weight gain (PWG), and specific growth rate (SGR) among all treatments. FBW, PWG, and SGR significantly decreased when dietary carbohydrate levels exceeded 240.0 g kg-1. Light microscopy results showed that dietary excessive carbohydrate levels will lead to vacuole, hepatopancreatic tubule lysis, and lipid droplets in hepatopancreas. Expression levels of genes involved into glucose transport, glucose and lipid metabolism, and insulin-like metabolism in the hepatopancreas were significantly influenced by dietary carbohydrate levels. In general, the results of present study demonstrated that dietary optimal carbohydrate levels could not only improve PWG and SGR but also influence the expression of genes involved in glucose, lipid, and insulin-like metabolism. The optimal actual carbohydrate level was estimated to be 87.8-98.4 g kg-1 based on broken-line and second-order polynomial regression analysis between PWG against dietary carbohydrate levels for juvenile Portunus trituberculatus.

Effect of dietary replacement of fish meal with low-gossypol cottonseed protein concentrate on growth performance and expressions of genes related to protein metabolism for swimming crab (Portunus trituberculatus)

An eight-week feeding trial was conducted to evaluate the effect of dietary replacement of fish meal with low-gossypol cottonseed protein concentrate (CPC) on growth performance, body composition, amino acid profiles in tissues, antioxidant capacity and the expression levels of genes related to protein metabolism and inflammation for juvenile swimming crab (Portunus trituberculatus). Six isonitrogenous and isolipidic diets (450 g kg−1 protein and 80 g kg−1 lipid) were formulated to contain graded levels of CPC, and fish meal was replaced with CPC at 0, 10%, 20%, 30%, 40% and 60% levels (CPC00, CPC10, CPC20, CPC30, CPC40 and CPC60), free gossypol concentrations in experimental diets were analyzed to be 0, 32.62, 66.10, 106.71, 133.13, 192.21 mg kg−1 respectively. Arginine, lysine and methionine were added to ensure consistency of these three essential amino acids in all experimental diets. Each diet was randomly assigned to triplicate groups of 30 juvenile swimming crabs (approximately initial weight 7.59 ± 0.06 g) that were stocked in rectangle plastic baskets. Results of present study indicated that crabs fed the CPC60 diet had the lower final weight (FW), percent weight gain (PWG), specific growth rate (SGR) and feed efficiency (FE) than those fed the CPC20 diet, and crabs fed the CPC20 and CPC30 diets had the highest survival among all treatments, and the lowest survival was occurred at crabs fed the CPC60 diet. The hepatopancreas color was blackened with the replacement levels of fish meal with CPC increasing from 0% to 60%. Free gossypol concentration in hepatopancreas significantly increased with increase of dietary replacement levels of fish meal with CPC. Crabs fed the CPC40 and CPC60 diets had lower lipid content in whole body than those fed the other diets. Arginine, lysine, methionine and isoleucine contents in muscle were significantly influenced by dietary replacement levels of fish meal with CPC. ALT, AST, TP, ALB, HDL and ALP in hemolymph, and T-AOC, ALT, AST, SOD, GSH, GSH-ST in hepatopancreas were significantly affected by dietary replacement of fish meal with CPC. Dietary replacement levels of fish meal with CPC could influence the expressions of genes related into protein metabolism such as tor, akt, s6, s6k1, 4ebp1, eif4e1a, eif4e2, eif4e3, antioxidant and immune such as gpx, cat, cMnsod and lysozyme, as well as growth such as tlr4, irak4, myd88, traf6, and inflammation such as relish. In conclusion, based on two slope broken-line and quadratic broken-line regression analysis of PWG against the dietary replacement of fish meal with CPC, the optimal replacement level of fish meal with CPC was estimated to be 19.07% and 22.73% for juvenile swimming crab, respectively. Results demonstrated that CPC could partially replace fish meal, but had no negative effects on growth performance, antioxidant activity and immunity of swimming crab.

Dietary manganese levels influence growth, manganese bioaccumulation and expression of genes involved in antioxidant response of swimming crab ( Portunus trituberculatus )

An 8-week feeding trial was conducted to investigate the effects of dietary manganese (Mn) levels on growth performance, Mn bioaccumulation in tissues and antioxidant capacity in juvenile swimming crab (Portunus trituberculatus). Six semipurified experimental diets were formulated to contain graded levels of Mn (manganese sulphate monohydrate as Mn source); the analysed Mn concentrations were 9.8, 20.2, 29.7, 38.2, 65.6 and 123.6 mg/kg. The percent weight gain (PWG) and specific growth rate (SGR) were significantly affected by dietary Mn levels; the highest PWG and SGR were found in crabs fed with the diet containing 38.2 mg/kg Mn. Crabs fed with the diet containing 123.6 mg/kg Mn had higher Mn concentration in muscle and carapace than those fed with the other diets. However, the highest Mn concentration in hepatopancreas was recorded in crabs fed with the diet containing 65.6 mg/kg Mn. Moreover, the lowest activities of manganese superoxide dismutase (Mn-SOD), catalase (CAT) and total antioxidant capacity (T-AOC) occurred at crabs fed with the dietary 9.8 mg/kg Mn. Crabs fed with the 9.8 mg/kg Mn diet exhibited lower relative expression of genes involved into antioxidant ability such as cytosolic manganese superoxide dismutase (cMnsod), glutathione peroxidase (gpx), peroxiredoxin (prx) and catalase (cat) in hepatopancreas than those fed with the other diets, and the highest expression were recorded in crabs fed with the diet containing 38.2 mg/kg Mn. Based on two slope broken-line and quadratic regression analysis of PWG against dietary Mn levels, the optimum dietary Mn requirement was estimated to be 48.02 and 53.30 mg/kg for juvenile Portunus trituberculatus. The present study provided further insight into the function of dietary Mn in crustacean.

Effect of dietary carbohydrate sources on the growth, glucose metabolism and insulin pathway for swimming crab, Portunus trituberculatus

An eight-week feeding trial was conducted to investigate the effects of dietary carbohydrate sources on growth performance, glycogen content in tissue, insulin signaling pathway, hepatic glucose transport and metabolism in juvenile swimming crab (Portunus trituberculatus). Four isonitrogenous (approximately 45.4% crude protein) and isolipidic (approximately 9.0% crude lipid) experimental diets were formulated to contain corn starch, wheat starch, dextrin and potato starch. Each diet was randomly assigned to triplicate groups of 30 juvenile swimming crabs (approximately 5.6 ± 0.1 g) that were stocked in rectangle plastic baskets. The results indicated that crabs fed with corn starch, wheat starch and dextrin diets had significantly higher final body weight (FBW), weight gain (WG), specific growth rate (SGR) and feed efficiency (FE) than those fed with potato starch diet. However, crabs fed with potato starch diet had the highest feed intake (FI) among all treatments. Crabs fed with corn starch, wheat starch and dextrin diets exhibited significantly higher glycogen in tissue and ATP content in hepatopancreatic than those fed with potato starch diet. The lowest concentration of insulin-like and hyperglycemic hormone were observed in crabs fed with potato starch diet. The lowest relative expression levels of PK, FBpase, PEPCK, GLUT1, GLUT2, GK, G6PDH, 6PGDH, ACC, IGF1R, PI3K and FOXO were occurred at crabs fed with dextrin diet. In conclusion, swimming crab could make better use of corn starch, wheat starch and dextrin, however, potato starch is not suitable as carbohydrate source. The results demonstrated that the ratio of amylose/amylopectin in diets was positively correlated with expression levels of genes related to glycolipid metabolism, glucose transport and insulin pathway.