The P4' Peptide-Carrying Bacillus subtilis in Cottonseed Meal Improves the Chinese Mitten Crab Eriocheir sinensis Innate Immunity, Redox Defense, and Growth Performance.

This study developed a recombinant Bacillus subtilis to carry the LGSPDVIVIR peptide (cmP4) isolated from the hydrolyzed products of cottonseed meal with excellent antioxidant and immune-enhancing properties in vitro. It was carried as a tandem of five cmP4 peptides (cmP4') to be stably expressed on a large scale. Then, its effectiveness was evaluated in Chinese mitten crab (Eriocheir sinensis) based on growth performance, redox defense, and innate immunity. A total of 280 crabs (mean body weight: 41.40 ± 0.14) were randomly assigned to seven diets including a control one (without B. subtilis) and six experimental ones with different doses (107,108, and 109 CFU/kg) of unmodified and recombinant B. subtilis, respectively, for 12 weeks. Each diet was tested in four tanks of crabs (10/tank). In terms of bacterial dosages, the final weight (FW), weight gain (WG), hemolymph and hepatopancreatic activities of superoxide dismutase (SOD), catalase (CAT), lysosome (LZM), acid phosphatase (ACP) and alkaline phosphatase (AKP), and hepatopancreatic transcriptions of cat, mitochondrial manganese superoxide dismutase (mtmnsod), thioredoxin-1 (trx1), and prophenoloxidase (propo) all increased significantly with increasing B. subtilis dosages, while hemolymph and hepatopancreatic malondialdehyde (MDA) content and the transcriptions of toll like receptors (tlrs), NF-κB-like transcription factor (relish), and lipopolysaccharide-induced TNF-α factor (litaf) all decreased remarkably. In terms of bacterial species, the recombinant B. subtilis group obtained significantly high values of FW, WG, hemolymph, and hepatopancreatic activities of SOD, CAT, LZM, ACP, and AKP, and the transcriptions of mtmnsod, peroxiredoxin 6 (prx6), and propo compared with the unmodified B. subtilis, while opposite results were noted in hemolymph and hepatopancreatic MDA content and the transcriptions of tlrs, relish, and litaf. These results indicated that dietary supplementation with 109 CFU/kg of recombinant B. subtilis can improve the growth performance, redox defense, and nonspecific immunity of E. sinensis.

Effects of a high-fat and high-carbohydrate diet on appetite regulation and central AMPK in the hypothalamus of blunt snout bream (Megalobrama amblycephala).

Adenosine monophosphate-activated protein kinase (AMPK) is a sensor of cellular energy changes and controls food intake. This study investigates the effect of a high-calorie diet (high fat diet [HFD], high carbohydrate diet [HCD] and high energy diet [HED]) on appetite and central AMPK in blunt snout bream. In the present study, fish (average initial weight 45.84 ± 0.07 g) were fed the control, HFD, HCD and HED in four replicates for 12 weeks. At the end of the feeding trial, the result showed that body mass index, specific growth rate, feed efficiency ratio and feed intake were not affected (p > 0.05) by dietary treatment. However, fish fed the HFD obtained a significantly higher (p < 0.05) lipid productive value, lipid gain and lipid intake than those fed the control diet, but no significant difference was attributed to others. Also, a significantly higher (p < 0.05) energy intake content was found in fish-fed HFD, HCD and HED than those given the control diet. Long-term HFD and HCD feeding significantly increased (p < 0.05) plasma glucose, glycated serum protein, advanced glycation end product, insulin and leptin content levels than the control group. Moreover, a significantly lower (p < 0.05) complex 1, 2 and 3 content was found in fish-fed HFD and HCD than in the control, but no differences (p > 0.05) were attributed to those in HED. Fish-fed HED significantly upregulated (p < 0.05) hypothalamic ampα 1 and ampα 2 expression, whereas the opposite trend was observed in the hypothalamic mammalian target of rapamycin than those in HFD and HCD compared to the control. However, hypothalamic neuropeptide y, peroxisome proliferator-activated receptor α (pparα), acetyl-coa oxidase and carnitine palmitoyltransferase 1 were significantly upregulated (p < 0.05) in the HCD group, while the opposite was seen in cholecystokinin expression compared to those in the control group. Our findings indicated that the central AMPK signal pathway and appetite were modulated according to the diet's energy level to regulate nutritional status and maintain energy homoeostasis in fish.

The Benefits of Nanosized Magnesium Oxide in Fish Megalobrama amblycephala: Evidence in Growth Performance, Redox Defense, Glucose Metabolism, and Magnesium Homeostasis.

This study evaluated the effects of dietary magnesium oxide nanoparticles (MgO NPs) on the growth, redox defense, glucose metabolism, and magnesium homeostasis in blunt snout bream. Fish (12.42 ± 0.33 g) were fed seven diets containing graded levels of MgO NPs (0, 60, 120, 240, 480, 960, and 1920 mg/kg) for 12 weeks. Whole-body Mg retention decreased significantly as the dietary Mg increased. As dietary MgO NPs levels reached 120 mg/kg, the growth performance and feed utilization remarkably improved. When added at 240 mg/kg, oxidative stress was significantly reduced evidenced by the increased Mn-sod transcription and the decreased CAT and GSH-Px activities and the MDA content. Meanwhile, it enhanced glucose transport, glycolysis, and glycogen synthesis, while inhibiting gluconeogenesis, as was characterized by the increased transcriptions of glut2, gk, and pk, and the decreased transcriptions of fbpase and g6pase. In addition, the supplementation of 120 mg/kg MgO NPs promoted Mg transport marked by a significant increase in the protein expressions of TRMP7, S41A3, and CNNM1. In conclusion, the moderate supplementation of MgO NPs improved the growth performance, reduced hepatic oxidative stress, and promoted glucose transport, glycolysis, glycogen synthesis, and magnesium homeostasis in fish while inhibiting glu.

Interactions between dietary carbohydrate and thiamine: implications on the growth performance and intestinal mitochondrial biogenesis and function of Megalobrama amblycephala.

A12-week experiment was conducted to evaluate the influences of thiamine ongrowth performance, and intestinal mitochondrial biogenesis and function of Megalobramaamblycephala fed a high-carbohydrate (HC) diet. Fish (24·73 (sem 0·45) g) were randomly assigned to one of four diets: two carbohydrate (CHO) levels (30 and 45 %) and two thiamine levels (0 and 1·5 mg/kg). HC diets significantly decreased DGC, GRMBW, FIMBW, intestinal activities of amylase, lipase, Na+, K+-ATPase, CK, complexes I, III and IV, intestinal ML, number of mitochondrial per field, ΔΨm, the P-AMPK: T-AMPK ratio, PGC-1ß protein expression as well as the transcriptions of AMPKα1, AMPKα2, PGC-1ß, mitochondrial transcription factor A, Opa-1, ND-1 and COX-1 and 2, while the opposite was true for ATP, AMP and reactive oxygen species, and the transcriptions of dynamin-related protein-1, fission-1 and mitochondrial fission factor. Dietarythiamine concentrations significantly increased DGC, GRMBW, intestinal activities of amylase, Na+, K+-ATPase, CK, complexes I and IV, intestinal ML, number of mitochondrial per field, ΔΨm, the P-AMPK:T-AMPK ratio, PGC-1ß protein expression as well as the transcriptions of AMPKα1, AMPKα2, PGC-1ß, Opa-1, ND-1, COX-1 and 2, SGLT-1 and GLUT-2. Furthermore, a significant interaction between dietary CHO and thiamine was observed in DGC, GRMBW, intestinal activities of amylase, CK, complexes I and IV, ΔΨm, the AMP:ATP ratio, the P-AMPK:T-AMPK ratio, PGC-1ß protein expression as well as the transcriptions of AMPKα1, AMPKα2, PGC-1ß, Opa-1, COX-1 and 2, SGLT-1 and GLUT-2. Overall, thiamine supplementation improved growth performance, and intestinal mitochondrial biogenesis and function of M. amblycephala fed HC diets.

Protective effects of dietary icariin on lipopolysaccharide-induced acute oxidative stress and hepatopancreas injury in Chinese mitten crab, Eriocheir sinensis.

To investigate the effects of dietary icariin (ICA) supplementation on acute oxidative stress and hepatopancreatic injury induced by lipopolysaccharide (LPS) injection in Eriocheir sinensis, an 8-week feeding trial of crabs was conducted using 4 diets with different supplementation levels of ICA (0, 50, 100, and 200 mg/kg diet weight, respectively), and then challenged with LPS of 400 µg/kg body weight for 6 h. Results showed that 100 mg/kg ICA supplementation increased the antioxidant capacity, reduced the stress-related indicators in haemolymph, strengthen the mitochondrial membrane potential, and reduce apoptosis compared to the single LPS-treated crabs. The expressions of apoptosis-related genes and proteins were also evaluated to further understand the effects of dietary ICA pretreatment on LPS-induced cell apoptosis. As a result, dietary 100 mg/kg diet weight ICA pre-addition significantly down-regulated the expression of HSP60, HSP70, Caspase 3c, Caspase 8, Caspase 3, Caspase 9, P38, and Bax (P < 0.05), and alleviated the suppressed expression of PI3K, AKT, MEK, and Bcl-2 (P < 0.05) in crabs challenged with LPS. Overall, this research reveals that ICA supplementation of 100 mg/kg diet weight could enhance the resistance to oxidative damage and apoptosis in E. sinensis facing LPS challenge.

Optimal Niacin Requirement of Oriental River Prawn Macrobrachium nipponense as Determined by Growth, Energy Sensing, and Glycolipid Metabolism.

Niacin is indispensable for the growth and development of aquatic animals. However, the correlations between dietary niacin supplementations and the intermediary metabolism of crustaceans are still poorly elucidated. This study explored the effects of different dietary niacin levels on the growth, feed utilization, energy sensing, and glycolipid metabolism of oriental river prawn Macrobrachium nipponense. Prawns were fed with different experimental diets containing graded niacin levels (15.75, 37.62, 56.62, 97.78, 176.32, and 339.28 mg/kg, respectively) for 8 weeks. Weight gain, protein efficiency, feed intake, and hepatopancreas niacin contents all maximized in the 176.32 mg/kg group with significance noted with the control group (P <0.05), whereas the opposite was true for feed conversion ratio. Hepatopancreas niacin concentrations increased significantly (P < 0.05) as dietary niacin levels increased, and peaked at the 339.28 mg/kg group. Hemolymph glucose, total cholesterol, and triglyceride concentrations all maximized in the 37.62 mg/kg group, while total protein concentration reached the highest value in the 176.32 mg/kg group. The hepatopancreas mRNA expression of AMP-activated protein kinase α and sirtuin 1 peaked at the 97.78 and 56.62 mg/kg group, respectively, and then both decreased as dietary niacin levels increased furtherly (P < 0.05). Hepatopancreas transcriptions of the genes related to glucose transportation, glycolysis, glycogenesis, and lipogenesis all increased with increasing niacin levels up to 176.32 mg/kg, but decreased significantly (P < 0.05) as dietary niacin levels increased furtherly. However, the transcriptions of the genes related to gluconeogenesis and fatty acid ß-oxidation all decreased significantly (P < 0.05) as dietary niacin levels increased. Collectively, the optimum dietary niacin demand of oriental river prawn is 168.01-169.08 mg/kg. In addition, appropriate doses of niacin promoted the energy-sensing capability and glycolipid metabolism of this species.

Effects of dietary xylooligosaccharide prebiotic supplementation on growth, antioxidant and intestinal immune-related genes expression in common carp Cyprinus carpio fed a high-fat diet.

This study investigated the effects of xylooligosaccharide (XOS) supplementation on growth, intestinal enzyme, antioxidant and immune-related genes in common carp Cyprinus carpio fed a high-fat diet (HFD). One hundred and ninety two fish with an initial weight of 19.61 ± 0.96 g were allocated into 24 tanks (eight fish per tank in four replicate) and were fed the control diet, HFD, HFD with 0.5%, 1%, 2% and 3% XOS supplementation. From the result, fish offered HFD with 1% XOS supplementation significantly obtained a higher body mass index and feed efficiency ratio, whereas condition factor was higher in fish fed HFD supplemented with 2% XOS but no difference was attributed to other supplemented group compared to control group. Also, fish fed HFD supplemented with 1%-2% XOS significantly improved protease, lipase, creatine kinase and sodium/potassium ATPase activities compared to other groups. Fish offered HFD were significantly lower in superoxide dismutase (SOD), catalase, glutathione peroxidase (GPX), myeloperoxidase, acid phosphatase, lysozyme activities and immunoglobulin content, but the opposite result was found for aspartate transaminase, alanine transaminase activities, malondialdehyde, protein carbonyl and cortisol content as compared with the control. However, this effect was reversed with HFD supplemented with XOS. Also, interleukin 1ß, interleukin 8, tumour necrosis factors, interferons, caspase-3 and caspase-9 in the intestine were all up-regulated in the HFD group, while the reverse pattern was found in SOD, GPX, lysozyme-C, complement 3 and mucin 5b (muc5b), than the control group. These effects were all enhanced by feeding the XOS diet, especially those fed 1%-3% supplementation. In conclusion, XOS inclusion can improve the growth, digestive enzymes, antioxidants and immune response of common carp fed HFD.

ß-Glucan Extracted from Highland Barley Alleviates Dextran Sulfate Sodium-Induced Ulcerative Colitis in C57BL/6J Mice.

Inflammatory bowel disease (IBD), which significantly affects human health, has two primary presentations: Crohn's disease and ulcerative colitis (UC). Highland barley is the most common food crop for Tibetans and contains much more ß-glucan than any other crop. Highland barley ß-glucan (HBBG) can relieve the gastrointestinal dysfunction and promote intestines health. This study aimed to evaluate whether HBBG can relieve UC in mice. A mouse model of UC was established by adding 2% dextran sulfate sodium (DSS) to drinking water for 1 week. UC was alleviated after the introduction of the HBBG diet, as indicated by reductions in the disease activity index (DAI) score, histopathological damage, and the concentration of colonic myeloperoxidase (MPO), along with an improvement in colonic atrophy. Furthermore, we found that HBBG can increase the relative transcriptional levels of genes encoding ZO-1, claudin-1, occludin, and mucin2 (MUC2), thereby reducing intestinal permeability. Additionally, HBBG maintained the balance of proinflammatory and anti-inflammatory cytokines and modulated the structure of the intestinal flora.

Dietary berberine can ameliorate glucose metabolism disorder of Megalobrama amblycephala exposed to a high-carbohydrate diet.

Blunt snout bream (Megalobrama amblycephala) were randomly assigned into three diets: normal-carbohydrate diet (NCD, 30% carbohydrate, w/w), high-carbohydrate diet (HCD, 43% carbohydrate), and HCB (HCD supplemented with 50 mg/kg berberine (BBR)). After 10 weeks' feeding trial, the results showed that higher levels of plasma glucose, triglyceride, and total cholesterol were observed in HCD-fed fish than in NCD-fed fish, while HCB feeding significantly ameliorated this effect. Moreover, HCB feeding remarkably reversed HCD-induced hepatic glycogen and lipid contents. In insulin signaling, BBR inclusion restored HCD-induced suppression of insulin receptor substrate mRNA expression and elevation of forkhead transcription factor 1 mRNA expression. In glucose metabolism, upregulated glucose transporter 2 and glycogen synthase mRNA expressions in the HCD group were observed compared to the NCD group. However, BBR adding reduced the mRNA expressions of glycogen synthase, phosphoenolpyruvate carboxykinase, and glucose-6-phosphatase and increased the transcriptional levels of glucose transporter 2 and pyruvate kinase. In lipid metabolism, BBR supplementation could reverse downregulated hepatic carnitine palmitoyl transferase I mRNA expression and upregulated hepatic acetyl-CoA carboxylase and fatty acid synthetase mRNA expressions in the HCD group. Taken together, it demonstrates that BBR could improve glucose metabolism of this species via enhancing liver's glycolysis and insulin signaling, while inhibiting liver's glycogen synthesis and gluconeogenesis. It also indicates that BBR could reduce the metabolic burden of the liver by inhibiting fat synthesis and promoting lipid decomposition, and then enhance fat uptake in peripheral tissues.

Dual protein ligand-modified gold nanoclusters for selective detection of serum sodium copper chlorophyllin.

Precisely tuning the emission wavelength of dual ligands co-functionalized gold nanoclusters (AuNCs) presents a bright prospect for highly selective drug detection. In this study, gold nanoclusters (AuNCs) with strong red fluorescence at 633 nm were synthesized using bovine serum albumin (BSA) and bromelain (Bro) as dual ligands for highly selective detection of sodium copper chlorophyllin (SCC). Interestingly, the auto-fluorescence interference of the dual ligands was effectively avoided due to the fluorescence emission red-shifts of the prepared BSA/Bro@AuNCs; this may be mainly due to the rigid structure of the proteins. The fluorescence of BSA/Bro@AuNCs was quenched after addition of SCC, which performed well and provided a good linear calibration curve in the 5.0-25.0 µM range (R2 = 0.999), and a detection limit as low as 0.5 µM. The proposed assay was further applied to the measurement of SCC in rat serums after an intravenous injection. The dual protein ligand-capped AuNCs showed great potential for drug analysis in actual biological samples.

Effect of γ-aminobutyric acid-rich yogurt on insulin sensitivity in a mouse model of type 2 diabetes mellitus.

This study aimed to investigate the effect of a γ-aminobutyric acid (GABA)-rich yogurt fermented with Streptococcus thermophilus fmb-5 on insulin sensitivity in high-fat and low-dose streptozotocin-induced type 2 diabetes mellitus mice. To study the ability of the yogurt to enhance insulin sensitivity, diabetic mice were treated with 0.5, 1, or 2 g/L of GABA yogurt once a day from wk 1 to 12. Compared with results in untreated diabetic mice, treatment with different dosages of GABA yogurt was associated with increased serum insulin and fat coefficient (fat weight relative to body weight) levels, decreased blood urea nitrogen, kidney coefficient (kidney weight relative to body weight), glucose area under the curve levels, and insulin sensitivity index, but did not alter blood glucose level or body weight. The highest dosage of GABA yogurt had a greater beneficial effect with respect to insulin resistance than the lower dosages. In particular, dietary supplementation of the high dosage of GABA yogurt favorably regulated HOMA-ß (homeostasis model assessment of ß-cell function), total cholesterol, high-density lipoprotein cholesterol, fat coefficient, and improved islet cells morphology. These results demonstrated that 2 g/L GABA yogurt could ameliorate insulin sensitivity. The GABA-rich yogurts appeared to be responsible for health-beneficial effects in this mouse model of diabetes.

Influences of glycyrrhetinic acid (GA) dietary supplementation on growth, feed utilization, and expression of lipid metabolism genes in channel catfish (Ictalurus punctatus) fed a high-fat diet.

An 8-week feeding trial was performed to test the effects of glycyrrhetinic acid (GA) on growth and some gene expression of hepatic lipid metabolism in channel catfish (initial body weight, 3.5 ± 0.02 g) fed high-fat diets. Fish were fed the control diet, high-fat diet (HFD), and HFD supplemented with 0.4, 0.8, and 1.2 mg/kg GA in 15 tanks at a stocking density of 21 fish/tank. Fish fed HFD were significantly lower in body weight gain and specific growth rate but higher in feed intake and feed conversion ratio in comparison to the control. Supplement of GA at 1.2 mg/kg remarkably improved these parameters as compared to the control diet. High levels of cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and low levels of high-density lipoprotein (HDL) in plasma were observed in fish fed HFD; the opposite was observed for fish fed HFD supplemented with GA. The transcription of fatty acid synthase (FAS), sterol regulatory element-binding protein-1 (SREBP1), liver X receptor alpha (LXRα), and hormone-sensitive lipase (HSL) was upregulated, while that of carnitine palmitoyltransferase I (CPT1), peroxisome proliferator-activated receptors alpha (PPARα), acyl-CoA oxidase (ACO), and microsomal triglyceride transfer protein (MTTP) mRNA expression were downregulated in fish fed HFD. The opposite was observed in fish fed HFD supplemented with GA as well as the control group. In conclusion, supplementing the HFD with GA at 1.2 mg/kg could improve the growth performance and lipid metabolism of channel catfish consuming HFD.

Dietary supplementation of Streptococcus faecalis benefits the feed utilization, antioxidant capability, innate immunity, and disease resistance of blunt snout bream (Megalobrama amblycephala).

This study aimed to investigate the effects of Streptococcus faecalis on the growth performance, intestinal histology, antioxidant capability, innate immunity, and disease resistance of blunt snout bream Megalobrama amblycephala. Fish were fed five experimental diets containing 0 (SF0, control), 1 × 105 (SF1), 1 × 106 (SF2), 1 × 107 (SF3), and 1 × 108 cfu/g (SF4) of Streptococcus faecalis, respectively, for 10 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila with the cumulative mortality recorded during a period of 96 h. The results showed that WG and FI of fish both showed no significant difference (P > 0.05) among all the treatments. However, the FCR was significantly (P < 0.05) affected by Streptococcus faecalis levels with the lowest value observed in the SF2 group, whereas the opposite was true for intestinal microvillus length (P < 0.05). Dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis significantly (P < 0.05) increased the hepatic activities of SOD, CAT, and GPx; plasma activities of LZM, MPO, ACP, and AKP; and the levels of C3, C4, and IgM of fish, compared with the control group. Similar results were also observed in the tissue expressions of Leap-I, Leap-II, muc2, and muc5b (P < 0.05), whereas the opposite was true for liver MDA contents and plasma NO levels (P < 0.05). At 96 h after challenge, the cumulative mortality of the control was significantly (P < 0.05) higher than that of the SF2 group, but it showed no statistical difference (P > 0.05) with that of the other treatments. These results indicated that dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis could not only improve the feed utilization of blunt snout bream but also enhance its antioxidant capability, innate immunity, and disease resistance.

Dietary fructooligosaccharide can mitigate the negative effects of immunity on Chinese mitten crab fed a high level of plant protein diet.

An 8-week feeding trial was carried out under controlled condition to evaluate the effect of dietary fructooligosaccharide (FOS) on growth performance, whole body composition, antioxidant status and immunity of crabs fed high levels of plant protein diets. Thus, six experimental diets were formulated (designated as F0P50, F0P60, F0P70, F0.2P50, F0.2P60 and F0.2P70), which contain two FOS levels (0 or 0.2%) and three plant protein levels (50, 60, or 70%) according to a 2 × 3 factorial design. The results showed that weight gain increased significantly as dietary plant protein level decreased from 70% to 50%. At 50% plant protein level, the addition of 0.2% FOS can significantly elevate weight gain (WG) (P < 0.05). The highest value in survival rate was observed in crabs fed F0.2P50 and F0.2P60 diet. Crabs fed F0.2P50 diet showed significantly higher crude protein content (P < 0.05) compared with those in other groups, but there were no significant differences in the contents of moisture, crude lipid and ash among all groups (P > 0.05). Catalase (CAT) activity in crabs fed F0.2P50 increased significantly (P < 0.05) compared with crabs fed F0P60, F0P70, F0.2P60 and F0.2P70, but malondialdehyde (MDA) concentrations decreased significantly (P < 0.05). Meanwhile, nitric oxide (NO) concentration, acid phosphatase (ACP) and alkaline phosphatase (AKP) activities of crabs fed 0.2% FOS diets increased significantly (P < 0.05) compared with crabs fed 0% FOS diets. The expressions of prophenoloxidase (propo) was significantly (P < 0.05) affected only by dietary plant protein levels with the highest values observed in 50% plant protein diet, whereas the opposite was true for Myeloid differentiation factor 88 (myd88). The mRNA expressions of mitochondrial manganese superoxide dismutase (mtmnsod), lipopolysaccharide-induced TNF-α factor (litaf) and toll like receptors (tlrs) were significantly affected (P < 0.05) by both FOS and plant protein levels. The cytosolic manganese superoxide dismutase (cytmnsod) mRNA expressions in F0.2P50 and F0.2P60 groups were significantly higher than those in F0P70 and F0.2P70 groups. The results in this study indicated that supplementation with 0.2% FOS can enhance growth performance in crabs fed lower plant protein diets and as well improve immunity in those fed with higher plant protein diets.

Benfotiamine, a Lipid-Soluble Analog of Vitamin B1, Improves the Mitochondrial Biogenesis and Function in Blunt Snout Bream (Megalobrama amblycephala) Fed High-Carbohydrate Diets by Promoting the AMPK/PGC-1ß/NRF-1 Axis.

This study evaluated the effects of benfotiamine on the growth performance and mitochondrial biogenesis and function in Megalobrama amblycephala fed high-carbohydrate (HC) diets. The fish (45.25 ± 0.34 g) were randomly fed six diets: the control diet (30% carbohydrate, C), the HC diet (43% carbohydrate), and the HC diet supplemented with different benfotiamine levels (0.7125 (HCB1), 1.425 (HCB2), 2.85 (HCB3), and 5.7 (HCB4) mg/kg) for 12 weeks. High-carbohydrate levels remarkably decreased the weight gain rate (WGR), specific growth rate (SGR), relative feed intake (RFI), feed conversion ratio (FCR), p-adenosine monophosphate (AMP)-activated protein kinase (AMPK)α/t-AMPKα ratio, peroxisome proliferator-activated receptor-γ coactivator-1ß (PGC-1ß) and nuclear respiratory factor-1 (NRF-1) protein expression, complexes I, III, and IV activities, and hepatic transcriptions of cytochrome b (CYT-b) and cytochrome c oxidase-2 (COX-2), whereas the opposite was true for plasma glucose, glycated serum protein, advanced glycation end product and insulin levels, tissue glycogen and lipid contents, hepatic adenosine triphosphate (ATP) and AMP contents and ATP/AMP ratio, complexes V activities, and the expressions of AMPKα-2, PGC-1ß, NRF-1, mitochondrial transcription factor A (TFAM), mitofusin-1 (Mfn-1), optic atrophy-1 (Opa-1), dynamin-related protein-1 (Drp-1), fission-1 (Fis-1), mitochondrial fission factor (Mff), and ATP synthase-6 (ATP-6). As with benfotiamine supplementation, the HCB2 diet remarkably increased WGR, SGR, tissue glycogen and lipid contents, AMP content, p-AMPKα/t-AMPKα ratio, PGC-1ß and NRF-1 levels, complexes I, III, IV, and V activities, and hepatic transcriptions of AMPKα-2, PGC-1ß, NRF-1, TFAM, Mfn-1, Opa-1, CYT-b, COX-2, and ATP-6, while the opposite was true for the remaining indicators. Overall, 1.425 mg/kg benfotiamine improved the growth performance and mitochondrial biogenesis and function in fish fed HC diets by the activation of the AMPK/PGC-1ß/NRF-1 axis and the upregulation of the activities and transcriptions of mitochondrial complexes as well as the enhancement of mitochondrial fusion coupled with the depression of mitochondrial fission.

The mechanism of action of a fat regulator: Glycyrrhetinic acid (GA) stimulating fatty acid transmembrane and intracellular transport in blunt snout bream (Megalobrama amblycephala).

High-fat diets are associated with fatty liver and aberrant hepatic lipid metabolism, and glycyrrhetinic acid (GA) has been shown to exert a beneficial effect on lipolysis and fat deposition in fish. In the present study, we evaluated the effect of GA on the growth performance and expression of hepatic lipid transport related genes in blunt snout bream (Megalobrama amblycephala) fed a high fat diet. Two hundred and sixteen fish (average body weight: 45.57 g ±â€¯0.98 g) were fed three experimental diets (6% fat/L6 group, control, 11% fat/L11 group, and 11% fat with 0.3 mg kg-1 GA/L11GA group) for 8 weeks. Compared to the control group, the weight gain and specific growth rate of high-fat fed group at the end of the trialwere significantly improved (P < .05).However, GA showed no effect on animals' growth performance(P > .05). Dietary supplementation with 0.3 mg kg-1 GA significantly decreased the hepatosomatic index, viscera/body ratio, and intraperitoneal fat ratio (P < .05), and up-regulated the expression levels of fatty acids transport protein (FATP), fatty acids binding protein (FABP), fatty acid translocase (CD36), carnitine palmitoyl transferase I (CPT1) and peroxisome proliferators-activated receptors α (PPARα) compared to both the L6 group and L11 group (P < .05). However, no significant difference was observed in fatty acid synthetase (FAS), acetyl-CoA carboxylase α (ACCα), or lipoprotein lipase (LPL) between groups (P > .05). In conclusion, GA significantly rescued high-fat diet induced hepatic lipid accumulation and metabolism dysfunction in M. amblycephalaby stimulating hepatic fatty acid transport and ß-oxidation. Dietary GA may be used as a promising supplement to alleviate high-fat diet induced side effects on M. amblycephala.

Lysine supplement benefits the growth performance, protein synthesis, and muscle development of Megalobrama amblycephala fed diets with fish meal replaced by rice protein concentrate.

This study aimed to investigate the effects of lysine supplement on the growth performance of blunt snout bream Megalobrama amblycephala fed diets with fish meal (FM) replaced by rice protein concentrate (RPC) with the potential mechanisms characterized. Fish were fed three diets, including the FM diet (containing FM), the RPC diet (FM replaced by RPC), and the MRPC diet (the RPC diet supplemented with lysine) for 8 weeks. Weight gain, protein efficiency ratio, and nitrogen and energy utilization of fish fed the FM diet were all significantly higher than those of the RPC treatment, but they showed no statistical difference with those of the MRPC group. Fish fed the RPC diet showed shorter villi length of the distal intestine than that of the other treatments. No significance was found in whole-body composition and intestinal and hepatic cell proliferation among all the treatments. However, fish fed the RPC diet obtained relatively low transcriptions of growth hormone (GH), GH receptor, insulin-like growth factor-I (IGF-I), target of rapamycin (TOR), ribosomal protein S6 kinase 1, myoblast determination protein, myogenic factor 5, and myostatin a (MSTNa) but high levels of eukaryotic translation initiation factor 4E-binding protein 2 (4E-BP2) than those of the other groups. Furthermore, little difference was found in the transcriptions of 4E-BP2, myogenin, muscle-specific regulatory 4, and MSTNb in muscle. Overall, these results showed that dietary supplement of lysine benefits the growth performance of blunt snout bream fed FM-free diets through the mediation of the GH-IGF-I axis, TOR signaling pathway, myogenic regulatory factors, and MSTN.

Effects of dietary glucose and starch levels on the growth, apparent digestibility, and skin-associated mucosal non-specific immune parameters in juvenile blunt snout bream (Megalobrama amblycephala).

A 10-week feeding trial was performed to evaluate the effects of different types and levels of carbohydrates in growth performance, apparent digestibility coefficients and skin-associated mucosal non-specific immune parameters in blunt snout bream (Megalobrama amblycephala). Fish were randomly fed four diets containing two carbohydrates (glucose and starch) diets and two carbohydrates levels (330 and 440 g kg-1). High carbohydrate levels remarkably increased the weight gain rate (WGR), apparent digestibility of dry matters, protein and carbohydrates, body crud protein content, plasma levels of aspartate transaminase (AST), and skin-associated mucosal levels of immunoglobulin M (IgM), HDL cholesterol, lysozyme (LZM), advanced the transcriptions of mucin 2 (Muc2), mucin 5b (Muc5b) and apolipoprotein A-I (apoA-I), whereas the opposite was true for feed conversion ratio (FCR), plasma levels of IgM, skin-associated mucosal levels of major histocompatibility complex (MHC) and ß-Defensins, and the transcriptions of heat shock protein 60 (Hsp60). In addition, carbohydrate types of glucose remarkably increased the survival rate, apparent digestibility of dry matters, protein and carbohydartes, body crud ash, plasma levels of total protein (TP), globulin (GLB), immunoglobulin M (IgM), complement C3 and complement C4 and the transcriptions of Muc5b. Whereas the carbohydrate types of starch remarkably increased viscerosomatic index (VSI), hepatosomatic index (HSI), condition factor (CF), abdominal fat percentage (AFP), apparent digestibility of liquid, advanced the transcriptions of Muc2, apoA-I and heat shock protein 70 (Hsp70). Significant interactions between different types and levels of dietary carbohydrates were also observed in WGR, apparent digestibility of dry matters, protein and liquid, body crud ash, plasma levels of TP, albumin (ALB) and AST, skin-associated mucosal levels of major histocompatibility complex (MHC) and ß-Defensins, and the transcriptions of Muc2 and Muc5b. Our results indicate that inclusion of high level of glucose in the diet of blunt snout bream could improve growth performance, nonspecific immunity, and increase the efficiency of protein, which is suggesting that high level of glucose could be used in feed production. However, the proportion of the specific formula of glucose using in feed needs further study.

Fishmeal replacement by rice protein concentrate with xylooligosaccharides supplement benefits the growth performance, antioxidant capability and immune responses against Aeromonas hydrophila in blunt snout bream (Megalobrama amblycephala).

This study aimed to investigate the effects of fishmeal (FM) replacement by rice protein concentrate (RPC) with a xylooligosaccharides supplement on the growth performance, antioxidant capability and immune response against Aeromonas hydrophila in blunt snout bream (Megalobrama amblycephala). Fish (46.85 ±â€¯0.34 g) were randomly assigned to one of 6 diets, namely the control diet (containing FM), the RPC diet (FM replaced by RPC) and RPC diet supplemented with 0.5, 1.5, 2.3 and 3% XOS respectively, for 8 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila for 96 h with the blood and liver sample obtained at 48 and 96 h respectively. The results showed that the final weight, weight gain and protein efficiency ratio of fish fed RPC diet were significantly (P < 0.05) lower than that of the control group, whereas the opposite was true for FCR. However, the supplement of 1.5% XOS remarkably (P < 0.05) improved these parameters compared to the control diet. Plasma total iron binding capacity of fish fed the RPC diet showed little difference (P > 0.05) with that of the control group, but it enhanced significantly (P < 0.05) with the supplement of 1.5% XOS compared to the control group. After bacterial infection, plasma lysozyme (LYM), complement 3, complement 4, myeloperoxidase (MPO), acid phosphatase (ACP) and alkaline phosphatase activities, as well as immunoglobulin M, levels all increased significantly (P < 0.05) with the maximum value is attained at 48 h, then they decreased significantly (P < 0.05) with further increasing time at 96 h. Similar results were also observed in liver superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities as well as malondialdehyde (MDA) content. Regarding dietary treatment, these parameters of fish fed RPC diet showed little difference (P > 0.05) with those fed the control diet but were significantly enhanced (P < 0.05) when RPC diet was supplemented with 1.5-2.3% XOS, except for hepatic (MDA) content which showed an opposite trend compared to the control group. After 96 h of challenge, the relative percentage survival (RPS) of fish fed XOS was significantly higher (P < 0.05) than that of fish offered the control and RPC diet. In addition, significant (P < 0.05) interactions between dietary XOS and sampling time were also observed in plasma LYM, MPO, ACP, and hepatic SOD, CAT, GPX, MDA, as well as RPS. In conclusion, this study indicated that dietary supplementation of 1.5%XOS could significantly improve the growth performance, antioxidant capability, innate immunity and A. hydrophila resistance of blunt snout bream fed diets with FM replaced by RPC.

Resveratrol supplementation improves lipid and glucose metabolism in high-fat diet-fed blunt snout bream.

Here, we aimed to investigate whether resveratrol (RSV) can ameliorate high-fat diet (HFD)-induced metabolic disorder in fish. Blunt snout bream (Megalobrama amblycephala) with average weight 27.99 ± 0.56 g were fed a normal fat diet (NFD, 5% fat, w/w), a HFD (11% fat), or a HFD supplemented with 0.04, 0.36, or 1.08% RSV for 10 weeks. As expected, fish fed a HFD developed hepatic steatosis, as shown by elevated hepatic and plasma triglycerides, raised whole body fat, intraperitoneal fat ratio and hepatosomatic index, and increased plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST). RSV supplementation lessened increases in body mass, whole body fat, and intraperitoneal fat, and alleviated development of hepatic steatosis, elevations of plasma triglyceride and glucose, and abnormalities of ALT and AST in HFD-fed fish. RSV supplementation increased SIRT1 messenger RNA (mRNA) expression and consequently hepatic mRNA expression of adipose triglyceride lipase (ATGL), carnitine palmitoyltransferase (CPT1a), and microsomal triglyceride transfer protein (MTTP), implying upregulation of lipolysis, ß-oxidation, and lipid transport, respectively, in the liver. Conversely, hepatic lipoprotein lipase (LPL), sterol regulatory element-binding protein 1 (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), and ATP citrate lyase (ACLY) mRNA expression were decreased, implying suppression of fatty acid uptake, lipogenesis, and fatty acid synthesis. Additionally, RSV downregulated glucokinase (GCK) and sodium-dependent glucose cotransporter 1 (SGLT1) and upregulated glucose transporter 2 (GLUT2) mRNA expression, thus restoring normal glucose fluxes. Thus, RSV improves lipid and glucose metabolisms in blunt snout bream, which are potentially mediated by activation of SIRT1.