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The study mainly aimed at the effects of dietary Senecio scandens buch-ham extract (SSBE) on the growth performance, body composition, plasma biochemical index, intestinal and liver histology and the expression of antioxidant, apoptosis and inflammatory related genes in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀). Basal diets supplemented with SSBE (10:1) 0%, 0.05%, 0.1%, 0.2% and 0.4% were fed hybrid grouper for 8 weeks. The results showed that WGR and SGR were significantly increased in the week 2 and week 4 in Diet 0.05% group (P < 0.05). The total protein, globulin and albumin significantly increased whereas alanine aminotransferase, triglyceride and alkaline phosphate in the plasma were significantly decreased in Diet 0.1% group (P < 0.05). The villi length, width, muscle thickness and the cross-sectional area of intestine were improved in Diet 0.05% and Diet 0.1% group. The expression levels of PPAR-α and CPT-1 in the liver of hybrid grouper were significantly increased following the supplementation of SSBE (P < 0.05). The expression levels of antioxidant related genes (CAT, GPX, GR and Keap1) and anti-inflammatory factor (IL-10) in liver, head kidney and spleen of hybrid grouper decreased significantly (P < 0.05). In addition, diets supplemented with 0.05%–0.1% SSBE had a good liver-protecting effect, but it would have a detrimental effect on hepatocytes when the content exceeds 0.2%. The above results indicated that the suitable additive amount of SSBE in hybrid grouper feed was 0.05%–0.1%. 1. Senecio scandens buch-ham extract (SSBE) could improve the WGR and SGR of hybrid grouper in the week 2 and week 4 at the group of Diet 0.05%. 2. SSBE could improve the morphology of liver and the hind gut of hybrid grouper when adding 0.05%–0.1% in the diet. 3. SSBE could decreased the expression of antioxidant related genes in liver, head kidney and spleen of hybrid grouper. 4. The appropriate dosage of SSBE in hybrid grouper feed is 0.05%–0.1%. [ABSTRACT FROM AUTHOR]

For thousands of years, leaves from the Ginkgo biloba tree have been a common treatment in Chinese medicine. The present study was conducted to investigate the effects of dietary ginkgo biloba leaf extract (GBE) supplementation on growth performance, plasma biochemical parameters, fish composition, immune responses, liver histology, and immune and apoptosis-related genes expression of hybrid grouper ( Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) fed high lipid diets. A basal diet supplemented with GBE at 0, 0.50, 1.00, 2.00, 4.00 and 10.00 g kg −1 was fed to hybrid grouper for 8 weeks. The study indicated that dietary GBE did not improve growth performance and feed utilization but it reduced intraperitoneal fat rate. There were no significant differences in condition factor, viscerosomatic index, hepatosomatic index, spleen index, relative gut length, food intake, protein deposit rate and survival among all groups ( P > 0.05). Dietary supplementation with 0.50–4.00 g GBE kg −1 diets effectively increased plasma HDL content and decreased plasma GLU, LDL and TG content in fish. Furthermore, dietary GBE had a significant effect on moisture, crude protein and lipid in the liver, and protein in the whole body of fish ( P < 0.05). Dietary supplementation with 0.50–1.00 g GBE kg −1 diets effectively decreased occurrence rates of the hepatocyte swelling, hepatocyte vacuolization, and nuclei shifting to the cellular periphery cytoplasmic vacuolization, meanwhile hepatic antioxidant enzymes (SOD, CAT and T-AOC) activities significantly increased whereas MDA content significantly decreased in fish fed diets supplemented with GBE ( P < 0.05). Moreover, dietary GBE up-regulated the expression of antioxidant genes (CAT, GPx and GR), immune-related genes (MHC2 and TLR3) and anti-inflammatory cytokines (IL-10 and TGF-β1), while dietary supplementation with 0.50–4.00 g GBE kg −1 diets down-regulated apoptosis-related genes (p53, caspase-9, caspase-8 and caspase-3) expression in the head kidney of hybrid grouper. These results indicated that hybrid grouper fed diets supplemented with GBE did not improve growth performance and feed utilization but it had hypolipidemic effects, improved hepatic antioxidant status, maintained normal liver histology and preserved liver function, increased immune-related genes expression and decreased apoptosis-related genes expression in the head kidney of hybrid grouper. [ABSTRACT FROM AUTHOR]

The study explored on the effects of dietary 0.4% dandelion extract on the growth performance, serum biochemical parameters, liver histology and the expression levels of immune and apoptosis-related genes in the head kidney and spleen of hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) at different feeding period. The results showed that the weight gain rate (WGR) of the hybrid grouper were significantly increased at the second and fourth weeks (P < 0.05), but there was no significant difference in WGR at the eighth week (P > 0.05). Compared with the control group, dietary dandelion extracts supplementation improve lipid metabolism, reduce lipid accumulation in liver and maintain normal liver histology at the second and fourth weeks. At the end of the second week, the relative expression levels of antioxidant related genes (MnSOD, GPX and GR) in the head kidney of hybrid grouper fed with dandelion extract increased significantly; at the end of week 4 and week 8, the relative expression levels of antioxidant related genes other than MnSOD did not change significantly. However, in the spleen of hybrid grouper, the expression of these antioxidant genes showed the opposite trend. At the end of the eighth week, dietary dandelion extract supplementation significantly increased the expression of inflammatory response related genes in head kidney of hybrid grouper, but showed the opposite trend in spleen. In conclusion, the short-term (2 or 4 weeks) application of 0.4% dandelion extract in feed had the effects of growth improvement, liver protection and immune stimulation on hybrid grouper due to its antioxidant and anti-inflammatory activities. The beneficial effect of dandelion extract on hybrid grouper was time-dependent, and its action time on different immune organs of hybrid grouper was not synchronous. • Short-term application of 0.4% dandelion extract (DE) in feed could significantly improve the weight gain rate of hybrid grouper. • Dietary DE supplementation could protect liver and reduce the accumulation of liver fat. • The beneficial effect of DE on hybrid grouper was time-dependent, and its optimum use time was 2–4 weeks. [ABSTRACT FROM AUTHOR]

The study explored on the effect of dietary compound plant extract supplementation on the growth performance, serum biochemical indicators, liver and intestinal morphological and gene expression levels in the head kidney and spleen of the hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀). The compound plant extracts (BDG) was a mixture of Bupleurum edulis extract, dandelion extract and Ginkgo biloba extract in a ratio of 1:4:1. Basal diets supplemented with BDG at 0, 0.75, 1.5, 3 and 6 g/kg were fed hybrid grouper for 8 weeks. The results showed that dietary 0.75 and 1.5 g/kg BDG supplementation could significantly increase the WGR and SGR of hybrid grouper (P < 0.05). And dietary 0.75 g/kg BDG could also significantly decrease serum aspartate aminotransferase, glucose and lactate dehydrogenase in hybrid grouper (P < 0.05). Dietary BGD supplementation protected the integrity of liver and intestinal morphological structure, reduced the accumulation of liver fat. Dietary BDG supplementation might enhance the immunity of hybrid grouper by regulating the expression of antioxidant and inflammation-related genes in head kidney and spleen of hybrid grouper. Our study demonstrated that the growth promoting effect of Bupleurum extract, dandelion extract and Ginkgo biloba extract in the ratio of 1:4:1 as a compound feed additive was better than any of them as a feed additive alone, and the dosage was less. The optimal additive dosage of BDG was 0.75 g/kg in hybrid grouper diets. • Dietary 0.75g/kg-1.5 g/kg BGD (compound plant extracts) could significantly improve the weight gain rate of hybrid grouper. • Dietary BGD supplementation could protect the integrity of liver and reduce the accumulation of liver fat. • The optimal additive dosage of BGD was 0.75 g/kg. [ABSTRACT FROM AUTHOR]

Excessive lipid accumulation and chemical abuse can induce fatty liver diseases in fish, but the underlying mechanism and therapies are unknown. The present study aims to evaluate the effects of Xiaochaihu Decoction (XCHD) on the growth performance, lipid metabolism and antioxidant function of hybrid grouper in vitro and in vivo , and provide evidence as to whether it can be potentially used as a medicine for liver diseases in aquaculture. In vitro , steatosis model of hybrid grouper primary hepatocytes were incubated for 48 h in control or lipid emulsion (LE)-containing medium with or without 24 h post-treatment with XCHD. XCHD treatment reversed the LE-induced intracellular lipid accumulation, cell viability and hepatocytes morphological structure. In vivo , a total of 300 hybrid grouper with an average initial weight of 25.43 ± 0.18 g were fed diets containing five graded levels of XCHD at 150–1200 mg/kg diet for 8 weeks. After that, a challenge trial was conducted by injection of D-GalN/LPS to induce liver injury. As a result, dietary supplementation with 150–300 mg/kg XCHD diets can significant improve growth performance and feed utilization (P < 0.05). Dietary XCHD down-regulated the expression of lipogenic-related genes (G6PD, DGAT2 and ME1) and up-regulated lipolysis-related genes (ATGL, PPARα and LPL) expression in the liver of hybrid grouper. Livers challenged with D-GalN/LPS exhibited extensive areas of vacuolization with the disappearance of nuclei and the loss of hepatic architecture. These pathological alterations were ameliorated by XCHD treatment. XCHD significantly down-regulated the D-GalN/LPS induced apoptosis-related genes caspase-3, caspase-9 and p53 mRNA expression and up-regulated the antioxidant-related genes CAT and MnSOD mRNA expression in dose dependent manner, respectively. XCHD potently reduced hepatic lipid accumulation and enhanced antioxidant capability in hybrid grouper and may be a potential fish-feed additive to prevent fatty liver diseases onset and progression. • In vitro ,XCHD have significantly hepatoprotective effects in fish. • In vivo , dietary supplemented of XCHD can promote growth and reduce lipid excess storage in fish. • XCHD enhanced antioxidant response to protect the liver injury in fish. [ABSTRACT FROM AUTHOR]

A 56-day feeding trial was conducted to evaluate the effects of replacing dietary fish meal (FM) with a complementary mixture of plant protein (PP) consisting of soybean meal (SBM), corn gluten meal (CGM) and cottonseed meal (CSM) on hepatic lipid metabolism and hepatocyte apoptosis in juvenile hybrid grouper, Epinephelus lanceolatus♂ × E. fuscoguttatus♀ (31.64 ± 0.82 g). A basal diet (FM60) with FM as the sole protein source was compared to diets progressively replacing 25% (FM45), 50% (FM30) and 75% (FM15) of FM protein. No significant differences were observed in growth performance and feed utilization when up to 75% of FM protein was replaced by PP sources. The hepatosomatic index (HSI) was markedly increased as dietary PP inclusion increased, but crude lipid content in the liver showed the opposite trend. Plasma cholesterol (CHO), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) of fish fed the PP containing diets were all significantly lower than the control. Liver histological analysis showed a higher levels of hepatocyte vacuolization and nuclear pycnosis occurred as dietary PP inclusion increased, but the amounts of hepatic lipid droplets showed a decreasing trend with increasing PP inclusion levels. Moreover, dietary PP inclusion down-regulated the mRNA levels of lipid metabolism-related genes including peroxisome proliferator-activated receptor alpha (PPARα), carnitine palmitoyltransferase 1 (CPT1), apolipoprotein AI (Apo-AI) and lipoprotein lipase (LPL). On the other hand, dietary PP sources also down-regulated the apoptosis-related genes including caspase-3, caspase-7, caspase-8 and p53. The present study provided new evidence for the PP sources-induced lipid metabolism in carnivorous fish, and provided new insight into the relationship between dietary PP sources and cell apoptosis. • This study provided new evidence for the plant protein sources-induced lipid metabolism in carnivorous fish species. • This study provided new insight into the relationship between dietary plant protein sources and cell apoptosis in fish. • This study showed that a combination of plant protein sources can replace up to 75% of fish meal protein in grouper feed. [ABSTRACT FROM AUTHOR]

Abstract In this study, two experiments were performed to explore the effect of Radix Bupleuri extracts (RBE) on growth, lipid deposition and metabolism and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) using in vitro and in vivo models. In vitro, we used 2 ml/L 20% lipid emulsion (LE)-induced steatosis in hybrid grouper primary hepatocytes, then RBE (200, 400 and 800 μg/ml) was added to the hepatocytes after (post-treatment) the incubation with 20% LE (2 ml/L) in the culture medium. We found that RBE markedly increased cell viability, which were consistent with hepatocytes morphological structure examination and lipid metabolism and immune related genes study. The above result suggested that RBE has a protective effect on this model of hepatocytes damage. In vivo, five graded levels of RBE at 0, 200, 400, 800 and 1600 mg/kg diet were supplemented to a basal diet with 15% lipid levels (high lipid), and fed to a total of 300 hybrid grouper with an average initial weight of 25.58 ± 0.05 g for 8 weeks. Growth performance, liver histology, plasma biochemical parameters, and expression of genes involved in lipid metabolism and immune-related were measured. The study indicated that dietary RBE significantly improved growth performance and feed utilization and reduced hepatosomatic index. Dietary supplementation with 200–800 mg/kg RBE diets effectively decreased serum ALP, ALT, AST and LDH contents in fish. Furthermore, adipogenesis relative mRNA levels of DGAT2, G6PD, ME1 and DGKα in fish fed 200–400 mg/kg RBE diets were lower (P < 0.05) than in those fed RBE0 diets, while dietary supplementation with 200–800 mg/kg RBE diets up-regulated lipolysis-related genes (CPT1, LPL and PPARα) expression in the liver of hybrid grouper. Moreover, dietary RBE down-regulated the expression of apoptosis-related genes (caspase-9), up-regulated the expression of antioxidant genes (CAT) and immune-related genes (MHC2, IKKα and TGF-β1). Thus, our data suggest that RBE suppressed lipid accumulation and enhanced immune capability in hybrid grouper both in vitro and in vivo. These results offer new insight into RBE as a hepatoprotective in fish. Highlights • In vitro and in vivo successfully established steatosis of hepatocytes or high-lipid dietsmodels in fish. • RBE at concentrations of 200–400 μg/ml has a protective effect on LE induced steatosis of hepatocytes. • Dietary administration of 400–800 mg/kg RBE can suppressed lipid accumulation and enhanced immune capability in fish. [ABSTRACT FROM AUTHOR]

Abstract The study investigated the effects of dietary supplementation with dandelion extracts (DE) on growth performance, feed utilization, body composition, serum biochemical, liver histology, immune-related gene expression and CCl 4 resistance of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀). A basal diet supplemented with DE at 0% (diet 0%), 0.1% (diet 0.1%), 0.2% (diet 0.2%), 0.4% (diet 0.4%) and 0.8% (diet 0.8%) were fed to hybrid grouper for 8 weeks. The results revealed that dietary DE had not a significant impact on growth performance and feed utilization (P > 0.05), but it could decrease the percent of crude lipids in whole body and increase the percent of crude protein in muscle (P < 0.05). Dietary DE increased the mRNA levels of antioxidant enzymes (catalase, glutathione peroxidase and glutathione reductase) and reduced inflammatory factor in the spleen and head-kidney of fish (P < 0.05), but reduced the expression of the liver antioxidant gene except for glutathione reductase (P < 0.05). Dietary supplementation with 0.2%–0.4% DE could effectively improve liver health. After injection of CCL 4 by 72 h, fish fed Diet0.2% and Diet0.4% showed regular hepatocyte morphology while fish fed Diet 0%, Diet 0.1% and Diet 0.8% showed hepatocyte damage. Higher survival rate and total blood cell count was observed in fish fed 0.1%–0.4% dietary DE (P < 0.05). In conclusion, DE could be used as a functional feed additive to enhance liver function of farmed fish. The best level of it should be between 0.2% and 0.4%. Highlights • DE could improve the antioxidant capacity of head kidney and spleen in hybrid grouper. • DE could protect the morphological integrity of hybrid grouper liver. • DE could improve the ability of hybrid grouper to resist CCl 4 stress. [ABSTRACT FROM AUTHOR]

Abstract The present study is aiming at evaluating the hepatoprotective of Radix Bupleuri extracts (RBE) on the d -galactosamine/lipopolysaccharide (D-GalN/LPS) induced liver injury of hybrid grouper in vitro and in vivo. In vitro, RBE (0, 200, 400 and 800 μg/ml) was added to the hybrid grouper primary hepatocytes before (pretreatment) the incubation of the hepatocytes with D-GalN (20 mM) plus LPS (1 μg/ml) in the culture medium. RBE at concentrations of 200, 400 and 800 μg/ml significantly improved cell viability and inhibited the elevation of TNF-α, IL-1β and IL-6 and significantly down-regulated the caspase-3, caspase-9 and P53 mRNA levels. In vivo administration of RBE at the doses of 0, 200, 400, 800 and 1600 mg/kg in the diet for 8 weeks prior to D-GalN (500 mg/kg) and LPS (20 μg/kg) intoxication. The study indicated that the RBE not only ameliorated liver injury, as evidenced by well-preserved liver architecture, but also significantly increased hepatic antioxidant enzymes activities in the D-GalN/LPS-induced liver injury animal model. Further demonstrating the protective effects of the RBE, we found that pretreatment with the RBE up-regulated the expression of antioxidant genes (GPx and MnSOD), while down-regulated apoptosis-related genes (caspase-3, caspase-9 and P53), immune related genes (MHC2 and TLR3) and pro-inflammatory cytokines (TOR and IKKα) mRNA expression in the liver of hybrid grouper. In brief, the present study showed that RBE can protect hepatocyte injury induced by D-GalN/LPS through elevating antioxidant enzyme activity and suppressing apoptosis and immune inflammatory responses. The results support the use of RBE as a hepatoprotective in fish. Highlights • In vitro and in vivo successfully established D-GalN/LPS induced hepatocytes or liver injury experimental models in fish. • Radix Bupleuri extracts (RBE) significantly improved cell viability and apoptosis from hepatocytes induced by D-GalN/LPS. • Diets with 400–800 mg/kg RBE improved hepatic antioxidant ability and immune responses induced by D-GalN/LPS in fish. [ABSTRACT FROM AUTHOR]