Influence of dietary zinc on growth, zinc bioaccumulation and expression of genes involved in antioxidant and innate immune in juvenile mud crabs (Scylla paramamosain)

The aim of present study was to investigate the effects of dietary Zn level on growth performance, Zn bioaccumulation, antioxidant capacity and innate immunity in juvenile mud crab (Scylla paramamosain). Six semi-purified diets were formulated to contain dietary Zn levels of 44.5, 56.9, 68.5, 97.3, 155.6 or 254.7 mg·kg-1, respectively. Dietary Zn level significantly influenced percent weight gain (PWG), with highest observed in crab fed the diet containing 97.3 mg·kg-1 Zn. Tissue Zn concentrations significantly increased as dietary Zn levels increased from 44.5 to 254.7 mg·kg-1. Retention of Zn in hepatopancreas increased with dietary Zn levels up to 68.5 mg·kg-1 and then significantly decreased. Moreover, inadequate dietary Zn (44.5 and 56.9 mg·kg-1) reduced anti-oxidation markers including total superoxide dismutase and copper/zinc superoxide dismutase activities and total anti-oxidant level. Crabs fed the diet with 44.5 mg·kg-1 Zn also showed significantly lower expression of genes involved in antioxidant status, such as Cu/Zn sod, glutathione peroxidase, catalase and thioredoxin than those fed diets containing 68.5 and 97.3 mg·kg-1 Zn. Highest activities of phenoloxidase and alkaline phosphatase were recorded in crab fed the diets containing 68.5 and 97.3 mg·kg-1 Zn. Expression levels of prophenoloxidase and toll-like receptor 2 were higher in crab fed the 97.3 mg·kg-1 Zn diet compared to crab fed the other diets. Based on PWG alone, the optimal dietary Zn level was estimated to be 82.9 mg·kg-1, with 68.5 to 97.3 mg·kg-1 recommended for maintaining optimal Zn bioaccumulation, oxidation resistance and innate immune response of juvenile mud crab

Effects of dietary lipid level on growth, fatty acid profiles, antioxidant capacity and expression of genes involved in lipid metabolism in juvenile swimming crab, Portunus trituberculatus

The regulation of lipogenesis and lipolysis mechanisms related to consumption of lipid has not been studied in swimming crab. The aims of present study were to evaluate the effects of dietary lipid levels on growth, enzymes activities, and expression of genes of lipid metabolism in hepatopancreas of juvenile swimming crab. Three isonitrogenous diets were formulated to contain crude lipid levels at 5.8 %, 9.9 % and 15.1 %, respectively. Crabs fed the diet containing 15.1 % lipid had significantly lower weight gain, specific growth rate and survival, and higher feed conversion ratio than those fed the 5.8 % and 9.9 % lipid diets. Crabs fed 5.8 % lipid had lower malondialdehyde concentrations in the hemolymph and hepatopancreas than those fed the other diets. Highest glutathione peroxidase in hemolymph and superoxide dismutase in hepatopancreas were observed in crabs fed 5.8 % lipid. The lowest fatty acid synthase and glucose 6-phosphate dehydrogenase activities in hepatopancreas were observed in crabs fed 15.1 % lipid, whereas crabs fed 5.8 % lipid had lower carnitine palmitoyltransferase-1 activity than those fed the other diets. Crabs fed 15.1 % lipid showed lower hepatopancreas expression of genes involved in LC-PUFA biosynthesis, lipoprotein clearance, fatty acid uptake, fatty acid oxidation, lipid anabolism and lipid catabolism than those fed the other diets, whereas expression of some genes of lipoprotein assembly and fatty acid oxidation were up-regulated compared with crabs fed 5.8 % lipid. Overall, high dietary lipid level can inhibit growth, reduce feed utilization and reduce antioxidant enzyme activities. Moreover, dietary lipid influenced enzyme activities and gene expression involved in lipid metabolism of juvenile swimming crab

Dietary DHA/EPA ratio affects growth, tissue fatty acid profiles and expression of genes involved in lipid metabolism in mud crab Scylla paramamosain supplied with appropriate n-3 LC-PUFA at two lipid levels

An 8-week feeding trial was conducted to determine the optimal dietary docosahexaenoic acid/eisosapentaenoic acid (DHA/EPA) ratio of mud crab (Scylla paramamosain) supplied with optimal n-3 LC-PUFA at two dietary lipid levels. Eight isonitrogenous diets were formulated to contain 7% and 12% crude lipid, each with DHA/EPA ratios of 0.6, 1.2, 2.3 and 3.2, respectively. Each diet was randomly assigned to triplicate groups of 30 juvenile mud crabs (initial weight 20.9 ± 0.6g) that were stocked in single crab cells. In crabs fed 7% lipid, the diet with a DHA/EPA ratio of 2.3 showed significantly higher weight gain than crabs fed the other ratios while in crabs fed 12% lipid, lower weight gain and specific growth rate were observed in crabs fed the diet with a DHA/EPA ratio of 0.6 than crabs fed the other ratios. Lipid content in hepatopancreas significantly increased as dietary DHA/EPA ratio increased from 1.2 to 2.3 in crabs fed 7% lipid, while no differences were observed among crabs fed the diets with DHA/EPA ratios higher than 0.6 when fed 12% lipid. Total fatty acid and DHA contents and DHA/EPA ratio showed increasing, and EPA decreasing, trends in muscle and hepatopancreas with increased dietary DHA/EPA ratio, at both dietary lipid levels. The hemolymph triacylglycerol and total cholesterol contents were higher in crabs fed dietary DHA/PA ratios of 1.2 and 2.3 than those fed ratios of 0.6 and 3.2 at 7% dietary lipid, and lowest low and high-density lipoprotein cholesterol contents were observed in crabs fed DHA/EPA dietary ratios of 0.6 and 3.2 at 7% and 12% lipid, respectively. The expression levels of fas, aco3 and fatp4 were significantly up-regulated, and cptⅠ, hsl and ldlr were down-regulated, with increased dietary DHA/EPA ratio in crabs fed 7% lipid. In crabs fed 12% lipid, the expression levels of g6pd, 6pgd, srebp-1, aco1 and fatp4 were down-regulated, and fabp-1 was up-regulated, with increased dietary DHA/EPA ratio. The expression levels of elovl4 and Δ6 fad initially increased and then decreased as dietary DHA/EPA ratio increased from 0.6 to 3.2 in crabs fed both 7% and 12% lipid. Based on analysis of weight gain versus dietary DHA/EPA ratio, the optimal dietary DHA/EPA ratios of mud crab S. paramamosa were estimated to be 2.2 and 1.2 when supplied with optimal n-3 LC-PUFA at 7% and 12% lipid, respectively

Dietary Betaine Mitigates Hepatic Steatosis and Inflammation Induced by a High-Fat-Diet by Modulating the Sirt1/Srebp-1/Pparɑ Pathway in Juvenile Black Seabream (Acanthopagrus schlegelii)

The present study aimed to elucidate the mechanism of dietary betaine, as a lipid-lowering substance, on the regulation of lipid metabolism and inflammation in juvenile black seabream (Acanthopagrus schlegelii) fed a high fat diet. An 8-week feeding trial was conducted in black seabream with an initial weight of 8.39 ± 0.01g fed four isonitrogenous diets including Control, medium-fat diet (11%); HFD, high-fat diet (17%); and HFD supplemented with two levels (10 and 20 g/kg) of betaine, HFD+B1 and HFD+B2, respectively. SGR and FE in fish fed HFD+B2 were significantly higher than in fish fed HFD. Liver histology revealed that vacuolar fat droplets were smaller and fewer in bream fed HFD supplemented with betaine compared to fish fed HFD. Betaine promoted the mRNA and protein expression levels of silent information regulator 1 (Sirt1), up-regulated mRNA expression and protein content of lipid peroxisome proliferator-activated receptor alpha (pparα), and down-regulated mRNA expression and protein content of sterol regulatory element-binding protein-1(srebp-1). Furthermore, the mRNA expression levels of anti-inflammatory cytokines in liver and intestine were up-regulated, while nuclear factor kB (nf-kb) and pro-inflammatory cytokines were down-regulated by dietary betaine supplementation. Likewise, in fish that received lipopolysaccharide (LPS) to stimulate inflammatory responses, the expression levels of mRNAs of anti-inflammatory cytokines in liver, intestine and kidney were up-regulated in fish fed HFD supplemented with betaine compared with fish fed HFD, while nf-kb and pro-inflammatory cytokines were down-regulated. This is the first report to suggest that dietary betaine could be an effective feed additive to alleviate hepatic steatosis and attenuate inflammatory responses in black seabream fed a high fat diet by modulating the Sirt1/Srebp-1/Pparɑ pathway

Dietary lipid and n-3 long-chain PUFA levels impact growth performance and lipid metabolism of juvenile mud crab, Scylla paramamosain

An 8 weeks feeding trial was conducted to evaluate the effects of dietary n-3 LC-PUFA levels on growth performance, tissue fatty acid profiles and relative expression of genes involved in lipid metabolism of mud crab (Scylla paramamosain). Ten isonitrogenous diets were formulated to contain five n-3 LC-PUFA levels at 7 % and 12 % dietary lipid levels. Highest weight gain and specific growth rate were observed in crabs fed the diets with 19.8 and 13.2 mg g-1 n-3 LC-PUFA at 7 % and 12 % lipid, respectively. Moisture and lipid contents in hepatopancreas and muscle were significantly influenced by dietary n-3 LC-PUFA at the two lipid levels. The DHA, EPA, n-3 LC-PUFA contents and n-3/n-6 PUFA ratio in hepatopancreas and muscle significantly increased as dietary n-3 LC-PUFA levels increased at both lipid levels. The expression levels of Δ6 FAD and ACO in hepatopancreas increased significantly, and expression levels of FAS, CPTⅠ and HSL were down-regulated, with increased dietary n-3 LC-PUFA regardless of lipid level. Based on weight gain, the n-3 LC-PUFA requirements of S. paramamosain were estimated to be 20.1 mg g-1 and 12.7 mg g-1 of diet at 7 % and 12 % dietary lipid, respectively. Over all, dietary lipid level influenced lipid metabolism, and purified, high-lipid diets rich in palmitic acid reduced the n-3 LC-PUFA requirement of juvenile mud crab

Dietary organic zinc promotes growth, immune response and antioxidant capacity by modulating zinc signaling in juvenile Pacific white shrimp (Litopenaeus vannamei)

An 8-week feeding trial was conducted to evaluate effects of dietary organic zinc (zinc amino acid chelate) on growth performance, mineral bioaccumulation in whole body, hepatopancreas and carapace, innate immune response and antioxidant capacity of juvenile Pacific white shrimp Litopenaeus vannamei. Five isonitrogenous and isolipidic diets were formulated to contain different zinc levels of 46.4 (basal diet), 65.5, 85.9, 108.4 and 130.6 mg kg−1. Dietary zinc level significantly influenced growth and feed utilization, with the lowest weight gain and highest feed conversion ratio observed in shrimp fed the basal diet. The optimal dietary zinc requirement was estimated to be 104.8 mg kg-1 for juvenile Pacific white shrimp. Shrimp fed the diet containing 130.6 mg kg−1 Zn had the highest zinc concentration in hepatopancreas and carapace, but there were no significant differences in calcium or phosphorus concentration in tissues. Dietary Zn increased the activities of lysozyme, alkaline phosphatase and polyphenol oxidase in hepatopancreas. Shrimp fed the diets supplemented with zinc had significantly higher activity of Cu/Zn SOD and lower content of malondialdehyde in hepatopancreas. The expression levels of toll, imd, lzm, proPO and alp involved in immunity and Cu/Zn sod related to oxidation resistance were up-regulated. Zinc also promoted the expression levels of mt and mtf-1, and up-regulated the expression of SLC39 family genes (zip3, zip9, zip11, zip14) in hepatopancreas. These data provided novel insights in the potential mechanism of organic zinc-induced enhancement of immunity and antioxidant capacity in Pacific white shrimp

Dietary soybean oil aggravates the adverse effects of low salinity on intestinal health in juvenile mud crab Scylla paramamosain

Salinity is one of the important factors affecting the physiological state of crustaceans in marine environments. Lipid plays major roles in energy supply and is main sources of essential fatty acids for membrane integrity, which is critical in adaptations to changes in salinity. Here we evaluated the effects of salinity (medium, 23 ppt and low, 4 ppt) and dietary lipid source (fish oil, FO and soybean oil, SO) on intestinal health of the marine crustacean mud crab Scylla paramamosain. The results indicated that low salinity and dietary SO (LSO group) significantly affected intestinal histomorphology, with a significant decrease of intestinal fold height and width as well as down-regulation of intestinal mRNA levels of tight junction genes compared to crab reared at medium salinity and fed FO diets (MFO group). Crabs reared at low salinity and fed SO showed an increased inflammatory response in intestine, which stimulated a physiological detoxification response together with apoptosis compared to crab in the MFO group. Low salinity and SO diets also could be responsible for multiply the pathogenic bacteria of Photobacterium and inhibit the beneficial bacteria of Firmicutes and Rhodobacteraceae in intestine, and act on a crucial impact on the development of intestinal microbial barrier disorders. The results of microbial function predictive analysis also support these inferences. The findings of the present study demonstrated that soybean oil as the main dietary lipid source could exacerbate the adverse effects of low salinity on intestinal health of mud crab, and provided evidence suggesting that dietary lipid source and fatty acid composition may play vital roles in intestinal health and the process of adaptation to environmental salinity in marine crustaceans

Litopenaeus vannamei BMAL1 Is a Critical Mediator Regulating the Expression of Glucose Transporters and Can Be Suppressed by Constant Darkness

Aryl hydrocarbon receptor nuclear translocator-like protein 1 (BMAL1) is a core circadian transcription factor that controls the 24-h cycle of physiological processes. In shrimp, the role of BMAL1 in the regulating glucose metabolism remains unclear. Firstly, we observed that the daily profile of BMAL1, GLUT1 and SGLT1 expression were synchronized in the intestine and the hepatopancreas of Litopenaeus vannamei. Then we examined the effects of BMAL1 on the gene expression of glucose transporter type 1 (SGLT1) and sodium-glucose cotransporter 1 (GLUT1) in vivo and in vitro. BMAL1 in L. vannamei shares 70.91–96.35% of sequence identities with other shrimp species and possesses the conserved helix-loop-helix domain and polyadenylation site domain. The in vitro dual-luciferase reporter assay and in vivo RNA interference experiment demonstrated that BMAL1 exerted a positive regulation effect on the expression of glucose transporters in L. vannamei. Moreover, we conducted an eight-week treatment to investigate whether light/dark cycle change would influence growth performance, and gene expression of BMAL1, GLUT1 and SGLT1 in L. vannamei. Our result showed that compared with natural light treatment, constant darkness (24-h darkness) significantly decreased (p < 0.05) serum glucose concentration, and suppressed (p < 0.05) the gene expression of BMAL1, GLUT1 and SGLT1 in the hepatopancreas and the intestine. Growth performance and survival rate were also decreased (p < 0.05) by constant darkness treatment. Our result identified BMAL1 as a critical mediator regulating the expression of glucose transporters, which could be suppressed by constant darkness in L. vannamei. It would be quite interesting to explore the mechanism of dark/light cycles on glucose transport and metabolism in L. vannamei, which might provide a feeding strategy for improving carbohydrate utilization in the future

Excessive dietary Cr6+ had no adverse effect on mud crab (Scylla paramamosain) but provoke consumers health risk

Hexavalent chromium (Cr6+) is a metallic contaminant that in organisms and poses a great threat to biological health and ecology due to its toxicity and biomagnification. However, the potential health risks to mud crabs and humans from dietary excessive Cr6+ were currently unknown. Therefore, a 9-week feeding trial was conducted to evaluate the toxic effects of dietary Cr6+ levels (1.4, 77.5 and 312.2 mg/kg) for mud crab (Scylla paramamosain). The results indicated that dietary Cr6+ levels had no significant effect on growth performance for mud crab. Cr concentrations in hepatopancreas and muscle significantly increased with dietary Cr6+ levels increasing from 1.4 to 312.2 mg/kg. The antioxidant capacity of mud crab was enhanced by increased the activities of superoxide dismutase (SOD) and scavenging capability for hydroxyl free radical (SCHR) to resist the Cr6+ toxicity. However, there was no statistical difference in the rate of apoptosis in hemocytes among all treatments. Expression levels of caspase 2 gene related to apoptosis in hemocytes significantly up-regulated with dietary Cr6+ level increasing from 1.4 to 312.2 mg/kg. For food safety, crabs fed diets with 77.5 and 312.2 mg/kg Cr6+ had target hazard quotient (THQ) values greater than 1 for the hepatopancreas, indicating a serious risk for human consumption. According to the target cancer risk (TCR) analysis, people who consumed muscle in mud crab fed diet with 312.2 mg/kg Cr6+ were at unacceptable cancer risk. Overall, the results of the present study indicated mud crab had a strong resistance to Cr6+ toxicity, and dietary excessive Cr6+ did not decline growth performance and survival, oxidative stress and apoptosis in mud crab. Excess Cr6+ were accumulated in tissues such as hepatopancreas and muscle, and consumption of crabs tissues fed diets with 77.5 and 312.2 mg/kg Cr6+ will provoke health and cancer risk in humans

Evaluation of cottonseed oil as a substitute for dietary fish oil of juvenile black seabream (Acanthopagrus schlegelii): Based on growth, lipid metabolism, antioxidant capacity and pi3k/akt pathway

An eight weeks feeding trial was conducted to evaluate the potential of cottonseed oil (CSO) as a substitute for fish oil (FO) on juvenile black seabream (Acanthopagrus schlegelii) (2.29 ± 0.01 g). Five diets were formulated with pure FO (FO) or partial CSO replacement of FO: 20% CSO (CSO20), 40% CSO (CSO40), 60% CSO (CSO60) and 80% CSO (CSO80), respectively. Results showed that fish fed CSO60 diet significantly increased growth performance compared to control group. Hepatic expression of genes related to long-chain polyunsaturated fatty acid biosynthesis were significantly up-regulated by dietary CSO60 compared to FO group. Hepatic paraffin section and serum biochemical indices showed that fish fed dietary CSO80 could increase vacuolar fat drops and cause liver damage. The expression levels of peroxisome proliferators-activated receptor alpha and its downstream lipolysis related genes were markedly up-regulated with the increasing of replacement levels of CSO, opposite results were recorded in sterol regulatory element-binding protein-1c and lipogenesis related genes. The malondialdehyde content in liver was decreased with the increasing replacement levels of CSO, whereas the antioxidant parameters were significantly higher in CSO60 group compared to FO group. Phosphatidylinositol-3 kinase and protein kinase B expression levels were significantly up-regulated by dietary CSO80, triggering inflammation by activating nuclear factor kappa b. Dietary CSO60 dramatically promote target of rapamycin expression level. With the increasing replacement levels of FO by CSO, the expression level of ribosomal protein S6 kinase 1 was significantly increased, whereas opposite result was found for the eukaryotic translation initiation factor 4E binding protein 2. Collectively, based on weight gain, double broken line model analysis showed that a maximum growth occurred at 59.13% replacement of dietary FO by CSO. These findings demonstrate that dietary CSO60 can improve growth and antioxidant capacity. However, dietary CSO80 can result in lipid deposition, liver injury and inflammation, hence, suppress growth performance