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1.
The intention of the study was to investigate the effect of ultrafiltered fish protein hydrolysate (UF) level on growth, feed utilization, apparent digestibility coefficients and proximal intestine peptide transporter 1 (PepT1) mRNA level for juvenile turbot (Scophthalmus maximus L.). Experimental diets (UF‐0, UF‐5, UF‐10, UF‐15 and UF‐20) were prepared containing about 68% plant protein, and fish meal protein was, respectively, replaced by 0%, 5%, 10%, 15% and 20% UF of dietary protein. Diet PP contained about 78% plant protein, and diet CAA contained about 10% crystalline amino acid mixture. All diets were fed to seven triplicate groups of turbot (initial weight 16.05 ± 0.03 g) for 68 days. Fish fed diet UF‐10 had an increasing tendency in growth compared with diets contained UF, while dietary UF level was not significantly correlated with specific growth rate and feed intake. Feed efficiency, protein efficiency ratio and protein productive value significantly correlated with dietary UF level, and fish fed diets contained low‐level UF had higher digestibility than that diets UF‐0, PP and CAA. There was a decreasing tendency in PepT1 expression level with dietary UF level. The results indicated that low‐level UF showed a positive effect on growth and feed utilization in juvenile turbot.  相似文献   

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Turbot aquaculture is a very important industry in China. However, it is hampered because of viral reddish body syndrome (VRBS) and high mortality caused by piscine turbot reddish body iridovirus (TRBIV). TRBIV virus is an icosahedron‐like and cytoplasmic DNA virus, belonging to Iridoviridae, Megalocytivirus. In previous studies, we have identified two antigen mimotopes using bioinformatics and constructed prokaryotic expression vectors. In this study, a fragment of major capsid protein (MCP) gene with the two antigenic epitopes was cloned into eukaryotic expression vector pVAX1, to generate a recombinant plasmid pVAX1‐TRBIV‐MCP. The plasmid DNA was transferred into turbot cell line TK using liposome, and transient expression was detected using RT‐PCR. After injection into turbot (Scophthalmus maximus), the expression of the antigen gene was analysed using RT‐PCR and was shown to express in all tested tissues in vaccinated fish 2 and 7 days post‐vaccination. The cumulative mortalities in the vaccinated and unvaccinated control fish were 30% and 88% respectively. Immune responses and upregulation of the expression of chemokine receptor, tumour necrosis factor, interferon and interferon‐induced antiviral molecules were observed in the vaccinated fish 60 h post‐vaccination. These results demonstrate that the vaccinated turbots had higher survival rate and produced specific serum antibodies following the TRBIV challenge. More studies are needed to develop and apply the promising DNA vaccine for virus control in turbot.  相似文献   

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A 10‐week feeding experiment in indoor flow‐through seawater system was conducted to investigate the effects of dietary krill hydrolysate on the expression of growth‐related genes in juvenile turbot (Scophthalmus maximus L.; initial body weight 9.45 ± 0.01 g). Three isonitrogenous and isolipidic experimental diets containing high plant protein were formulated to contain 0 (control), 50 g/kg (LKH) and 100 g/kg (HKH) krill protein hydrolysate (KH) to replace fishmeal, respectively. Triplicate groups of 30 fish were fed for 10 weeks to apparent satiation twice daily. At the end of the feeding trial, the mRNA expressions of insulin‐like growth factor (IGF‐1) gene in liver, peptide transporters (PepT1) gene in pyloric caeca and proximal intestine and neuropeptide Y (NPY) gene in brain in all groups were determined. IGF‐1, PepT1 and NPY expression levels in HKH group were significantly increased compared with those of LKH and control (< 0.05), which was consistent with the SGR, feed efficiency, PER and PPV. These results indicated that dietary 100 g/kg krill hydrolysate could improve growth performance and upregulate the mRNA expression of IGF‐1, PepT1 and NPY genes in juvenile turbot.  相似文献   

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The effects of fish protein hydrolysate (FPH) on growth, peptide and amino acid (AA) transporters, postprandial free AA and related gene expression of IGF‐1/AKT pathway were evaluated in turbot (Scophthalmus maximus). Three diets were formulated to contain the same low level of fishmeal; meanwhile 0, 45 and 180 g/kg FPH were, respectively, supplemented to the FF (FPH‐free), FL (FPH‐Low) and FH (FPH‐High) diets. Fish fed the FH diet improved the growth compared with the other groups. For peptide and AA transporters, PepT1, B0AT1, CAT1 and PAT1 mRNA levels in proximal or distal intestine decreased in fish fed the FH diet. The concentration of free total essential AAs in serum was higher in the FH treatment than that in the FF treatment at 2 and 6 hr after feeding. For IGF‐1/AKT pathway in muscle, IGF‐1, 4E‐BP1 and FoxO1 mRNA levels were the highest in the FH group, whereas IGF‐1R mRNA levels were the highest expression level in the FF group. In conclusion, down‐regulated AAs transport, alleviated the delayed postprandial peak of serum‐free AAs and increased muscle protein synthesis were observed to improve the growth when turbot was fed high FPH level diets containing a high plant protein.  相似文献   

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A 9‐week feeding trial was conducted using triplicate groups of turbot (6.50 ± 0.01 g) to explore the potential effects of silymarin. Three concentrations of silymarin (100, 200, and 400 mg/kg) were added to the plant protein‐based diet. Fish were randomly distributed into fiberglass tanks (30 fish per tank). The results showed that adding 100 mg/kg silymarin significantly improved the growth performance, with no effects on feed utilization. The antioxidant capacity in the liver was significantly improved in the 100 mg/kg and 200 mg/kg silymarin groups by not only inducing the activities of superoxide dismutase (SOD) and catalase but also increasing the messager RNA (mRNA) expression levels of SOD, glutathione peroxidase, and peroxiredoxin 6. Meanwhile, supplying 100 and 200 mg/kg of silymarin enhanced the heights of villi and enterocytes. Silymarin supplementation reduced the mRNA expression of interleukin‐8 and tumor necrosis factor‐α but induced the expression of transforming growth factor‐β (TGF‐β) in the intestine. These results indicated that silymarin was a potential nutraceutical that could enhance the growth performance and health status of turbot fed in a high plant protein diet. Adding 100 mg/kg silymarin to the plant protein diet achieved optimal performance in turbot.  相似文献   

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刘晋士  卫育良  徐后国  朱永祥  梁萌青 《水产学报》2023,47(4):049611-049611
为探究大菱鲆对不同来源水解蛋白的利用效率,实验选取太平洋狭鳕和牛蛙下脚料为蛋白来源,分别制备水解鱼蛋白和水解牛蛙蛋白,以初始体重为(8.00±0.01) g的大菱鲆为研究对象,进行为期56 d的养殖实验。实验设2个对照组,正对照组(PC)鱼粉含量为35.0%,负对照组(NC)鱼粉含量为26.5%;设2个实验组,水解鱼组(FPH)为26.5%的鱼粉和8.0%的水解鱼蛋白,水解牛蛙组(BPH)为26.5%的鱼粉和9.5%的水解牛蛙蛋白。结果显示,FPH组的终末体重、增重率和特定生长率显著高于BPH组和NC组,与PC组无显著差异。摄食6 h后,食糜必需氨基酸中赖氨酸、精氨酸、苏氨酸和缬氨酸在BPH组的含量显著高于PC组;多数非必需氨基酸在BPH组含量最高,但无显著差异。质子偶联氨基酸转运载体PAT1和小肽转运载体PepT1的mRNA表达量分别在FPH组和BPH组都显著高于PC组和NC组;碱性氨基酸转运载体CAT1和y+L型氨基酸转运载体y+LAT2的mRNA表达量在各处理组中无显著差异。研究表明,在饲料中添加鳕和牛蛙蛋白水解物均能提高大菱鲆的生长性...  相似文献   

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本研究利用低分子水解鱼蛋白设计了4组等氮等能的高植物蛋白饲料,研究不同水平低分子水解鱼蛋白对大菱鲆(Scophthalmus maximus L.)幼鱼[(4.16±0.01)g]生长性能、鱼体组成及肝脏中类胰岛素生长因子Ⅰ受体(Insulin-like growth factor receptor,IGF-IR)表达的影响.水解鱼蛋白分别替代总蛋白的5%(UF-5)、10%(UF-10)、20%(UF-20)的鱼粉,无添加FPH组为对照组(UF-0),用这4种饲料饲喂大菱鲆幼鱼84 d,结果显示,UF-0、UF-5和UF-10组的增重率、特定生长率无显著差异(P>0.05),但显著高于UF-20组(P<0.05);UF-0、UF-5组的饲料效率、蛋白质效率和蛋白质沉积率无显著差异(P>0.05),而显著高于UF-10、UF-20组(P<0.05);UF-0、UF-5和UF-10三组鱼体粗蛋白、粗脂肪含量无显著差异(P>0.05),但显著高于UF-20组(P<0.05);UF-5组必需氨基酸含量及必需氨基酸与非必需氨基酸比值显著高于其他3组(P<0.05),其他3组间无显著差异(P>0.05);肝脏中IGF-IR mRNA的表达随着水解鱼蛋白替代水平的增加而升高,且UF-20组与其他3组差异显著(P<0.05).结果表明,适当添加低水平水解鱼蛋白(UF-5)可促进大菱鲆幼鱼的生长、提高饲料效率及促进肌肉必需氨基酸的积累;高水平添加低分子水解鱼蛋白(UF-20)会抑制其生长及饲料利用等;低分子水解鱼蛋白可提高大菱鲆肝脏中IGF-IR基因的表达量.  相似文献   

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The study aimed to investigate the effects of crystalline Lys and Leu, Lys‐Leu dipeptide and Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide, on growth, postprandial Lys and Leu concentrations, gene expression of peptide and amino acid (AA) transporters and related gene expression of protein synthesis pathway in turbot. Three diets (CAA, Di and Tri) contained Lys and Leu as free (Lys and Leu), dipeptide (Lys‐Leu) and tripeptide (Lys‐Leu‐Lys/Leu‐Lys‐Leu) forms, respectively. Improved growth was observed in the Di group compared with the CAA group. For peptide and AAs transporters, PepT1, B0AT1 and y+LAT2 mRNA levels were affected in proximal intestine by dietary treatments. Free Lys and Leu concentrations in the CAA group were significantly higher than that of the Di and Tri groups at 6 hr postfeeding in serum and at 2, 6 and 24 hr postfeeding in muscle. For protein synthesis pathway in muscle, Akt2, TOR and S6k1 gene expression were the highest in the Di group and the lowest in the CAA group, whereas MuRF1 relative expression was the highest in the Tri group. In conclusion, dietary Lys‐Leu dipeptide was utilized more efficiently than free Lys and Leu or Lys‐Leu‐Lys/Leu‐Lys‐Leu tripeptide for turbot by regulating AAs transport, postprandial AAs concentration and the synthesis of muscle proteins.  相似文献   

11.
MicroRNA(miRNA)是一类内源性的非编码RNA,参与调节病毒复制、细胞增殖或凋亡以及肿瘤发生等生物反应。本研究探讨了miR-130c-5p在乌鳢水泡病毒(snakehead vesiculovirus, SHVV)感染中潜在靶基因g的靶向关系以及对病毒复制的影响。本研究以斑点叉尾鮰卵巢(Channel catfish ovary, CCO)为实验材料,通过qRT-PCR和Western blot测定SHVV不同感染时间和感染剂量条件下,病毒基因水平和蛋白水平以及miR-130c-5p变化情况。此外,将SHVV的g基因上miR-130c-5p对应的靶序列克隆到质粒pmirGLO,构建质粒pmirGLO-G用于双荧光素酶报告实验进行靶基因验证。结果显示,随着SHVV感染时间及剂量的不断增加,miR-130c-5p和g基因的表达水平都显著上调。进一步实验证明,miR-130c-5p类似物和pmirGLO-G质粒共转染可显著抑制荧光素酶活性强度,而转染miR-130c-5p抑制剂则明显上调了pmirGLO-G报告载体的荧光信号。此外,miR-130c-5p的过表达显著降低了病毒G基因的mRNA及蛋白表达,而抑制miR-130c-5p的表达则上调了g基因的mRNA及蛋白的表达水平。研究结果表明,miR-130c-5p通过靶向SHVV的g基因,引起G蛋白的降解,从而抑制SHVV的增殖。本研究结果为理解microRNA调控SHVV的致病机制提供了重要基础,为抗SHVV疫苗等药物的研发提供了理论支持。  相似文献   

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An experiment was conducted to evaluate the effects of five attractants on feed intake, growth performance and appetite‐regulating genes expression for juvenile turbot (Scophthalmus maximus L.). Fish were fed with seven diets: fish‐meal‐based diet (FM), fermented soybean meal replacing 45% fish meal (FSBM) and other diets with 1% compound attractant A (CA‐A) consisting of amino acids, compound attractant B (CA‐B) consisting of quaternary ammonium bases, compound attractant C (CA‐C) consisting of nucleotides, compound attractant G (CA‐G) consisting of the combination of CA‐A, CA‐B and CA‐C and blue mussel protein hydrolysate (BMH) on the basis of FSBM diet respectively. Daily feed intake in CA‐A group was significantly higher, but that in CA‐B group was significantly lower than FSBM group. Similarly, specific growth rate was significantly enhanced in CA‐A and CA‐G groups and were significantly decreased in CA‐B group. Protein efficiency ratio and protein retention in CA‐B group were significantly lower than FSBM group. Furthermore, BMH supplement increased the mRNA expression of Ghrelin in the intestine of fish. These results showed that CA‐A and CA‐G supplement could enhance growth in plant‐based diets, but CA‐B supplement had the opposite effect, which might be associated with lower feed intake and lower feed utilization.  相似文献   

16.
This study investigates the susceptibilities of the SPB cell line to fish viruses including giant seaperch iridovirus (GSIV‐K1), red sea bream iridovirus (RSIV‐Ku), grouper nervous necrosis virus (GNNV‐K1), chum salmon reovirus (CSV) and eel herpesvirus (HVA). GSIV‐K1, RSIV‐Ku and CSV replicated well in SPB cells, with a significant cytopathic effect and virus production. However, the cells were HVA and GNNV refractory. To examine the ability of SPB cells to stably express foreign protein, expression vectors encoding GNNV B1 and B2 fused to enhanced green fluorescent protein (EGFP) and GSIV ORF35L fused to DsRed were constructed and introduced by transfection into SPB cells. Stable transfectants displayed different morphologies compared with SPB and with each other. EGFP‐B1 was predominantly localized in the nuclei, EFPF‐B2 was distributed throughout the cytoplasm and nucleus, and granular 35L‐DsRed was localized with secreted vesicles. The expression of EFPF‐B2 in SPB cells produced blebs on the surface, but the cells showing stable expression of EGFP, EGFP‐B1 or 35L‐DsRed showed normal morphologies. Results show the SPB cells and the transfected cells grow well at temperatures between 20 and 35 °C and with serum‐dependent growth. SPB cells are suitable for studies on foreign protein expression and virology.  相似文献   

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Intraperitoneal injection of turbot with Cd induced the synthesis of a low molecular weight hepatic Cd-binding protein and a 500bp mRNA, which hybridised to a plaice metallothionein (MT) cRNA probe. The Cd-binding protein displayed cross-reactivity in a competitive ELISA with antiserum raised against rainbow trout MT and had the characteristic amino acid composition, metal stoichiometry and spectral characteristics of a Class I MT. Only one isoform was apparent on ion exchange chromatography. Southern blot analysis of DNA cleaved with four restriction enzymes suggested that only a single MT gene is present in turbot.In an established turbot fibroblast cell line, Cd induced MT mRNA and MT levels in a dose and time-dependent manner. MT was also induced by Cu, Hg and Zn but not Pb exposure. Physiological concentrations of glucocorticoids and sex hormones did not induce MT synthesis, although at high concentrations a positive response to corticosterone, dexamethasone, hydrocortisone or progesterone was observedin vitro indicating the possible presence of a functional steroid regulatory element in the fish MT gene.Abbreviations used AMP adenosine monophosphate - CHSE chinook salmon embryo - DMSO dimethyl sulphoxide - HPLC high performance liquid chromatography - KB kenacid blue - MT metallothionein - Mr molecular weight (kDaltons) - NR neutral red - PEG polyethylene glycol - RTH rainbow trout hepatoma - SDS sodium dodecyl sulphate - SSC 0.15M NaCl, 0.015M sodium citrate - TF turbot fibroblast  相似文献   

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D. Xu  G. He  K. Mai  Q. Wang  M. Li  H. Zhou  W. Xu  F. Song 《Aquaculture Nutrition》2017,23(5):1169-1178
The objective of the study was to evaluate the effects of incorporating plant protein blend in juvenile turbot (Scophthalmus maximus L.) diet on free amino acid (AA) concentration and the expression of genes related to peptide and AA transporters, key enzymes of AA metabolism and AA response (AAR) pathway. Fish were fed diets with fish meal (FM), or 400 g/kg FM replacement by plant protein blend for 9 weeks. Compared with the FM diet, PP40 diet did not affect plasma essential amino acid (EAA) concentration or AA metabolic enzymes gene in intestine, while it significantly upregulated all the detected peptide and neutral AA transporters gene. Results in muscle indicated that PP40 diet led to a great reduction of EAA concentrations and mRNA abundance of two kinds of AA transporters (SNAT2 and b0,+AT), while it greatly increased the gene expression of L‐type and T‐type AA transporters (LAT2 and TAT1) and the enzymes of AA catabolism (BCKDH‐E2) and anabolism (asparagine synthetase). In addition, the expression of genes related to AAR pathway were all greatly stimulated by PP40 diet in muscle. Our results provide a molecular explanation for the change of tissues AA concentrations caused by plant protein in turbot, which maybe applicable for general carnivorous fish.  相似文献   

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为研究饲料中不同水平的花生四烯酸(arachidonic acid,ARA)对发育前期的大菱鲆亲鱼性类固醇激素合成量及合成过程的影响,配制3种等氮等脂(脂肪含量13%)的实验饲料,分别含有不同梯度水平的花生四烯酸:0.72%(不添加花生四烯酸精制油的对照组,C),5.63%(添加低水平ARA的处理组,ARA-L)及15.03%(添加高水平ARA的处理组,ARA-H)(各数值均为占总脂肪酸的比例)。每组饲料投喂3个实验桶,每桶放25尾3龄大菱鲆亲鱼(雌雄比例约为1:1)。养殖实验在室内流水系统内进行,每天饱食投喂2次,养殖周期为5个月。养殖结束后,分别取性腺发育前期的雌雄鱼测血清雌二醇和睾酮的含量并检测性腺中性类固醇激素合成相关蛋白质的基因表达量。结果显示:与对照组相比,饲料中高水平的花生四烯酸显著降低了雌鱼血清中雌二醇的含量,而雄鱼血清中睾酮的含量在低水平花生四烯酸处理组显著降低。在卵巢中,饲料中高水平的花生四烯酸显著降低了促卵泡激素受体mRNA的表达量,但是饲料中低水平的花生四烯酸显著提高了17α羟化酶的mRNA表达量。在精巢中,饲料中低水平的花生四烯酸显著降低了固醇合成急性调节蛋白以及17α羟化酶的mRNA表达量,但显著升高了芳香化酶的mRNA表达量。饲料中花生四烯酸提高了性腺、肝脏和肌肉组织中花生四烯酸的含量,但降低了二十碳五烯酸(EPA)的含量,卵巢中的花生四烯酸累积量高于精巢。综上所述,饲料中添加一定量的花生四烯酸抑制了发育前期大菱鲆亲鱼雌二醇和睾酮的合成,在卵巢中,这种抑制作用可能是通过抑制促卵泡激素受体的表达来实现,而在精巢中,可能是通过抑制固醇合成急性调节蛋白以及17α羟化酶来实现。  相似文献   

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