鳜UCP1,2基因结构、序列分析及组织表达

从分子水平上探讨了鳜解偶联蛋白1、2 (UCP1, 2)基因结构、组织表达水平及与产热、脂肪代谢等生理机能的关系。通过与脊椎动物UCP1、UCP2基因序列进行比对,设计简并引物与特异引物进行PCR和RACE扩增、测序、拼接序列,获得UCP1、UCP2的基因组序列和内含子/外显子结构。基因组步行法克隆鳜肝脏UCP1、UCP2 基因5′侧翼序列。应用半定量RT-PCR的方法,以β-肌动蛋白作为外参照,在其指数期增长的范围内得到鳜不同组织UCP1、UCP2的相对表达水平。结果表明,UCP1基因组全序列为3 146 bp, 5′侧翼调控区为1 333 bp,含有5个内含子和6个外显子,开放阅读框(ORF)长942 bp,编码一个大小为313个氨基酸的蛋白质。UCP2基因组全序列为2 890 bp, 5′侧翼调控区为1 800 bp,含有7个内含子和8个外显子,ORF长939 bp,编码一个大小为312个氨基酸的蛋白质。UCP1、UCP2间隔外显子的内含子皆符合“GT-AG”规则,内含子的数目与哺乳动物一致。系统进化分析表明,鳜UCP1、UCP2氨基酸序列分别与鱼类UCP1、UCP2氨基酸序列聚为一支,且与UCP3、UCP4、UCP5分支区分明显。鳜UCP1、UCP2基因不同组织表达水平的高低可能与鳜本身的生态习性及各器官在产热、脂质代谢中的作用相关,但明确的分子机制尚待进一步研究。     

鳜胃蛋白酶原基因cDNA全长的克隆与序列分析

利用RT–PCR和cDNA末端快速扩增法(RACE)克隆鳜(Siniperca chuatsi)胃蛋白酶原基因cDNA全长序列，并对该基因的结构特征和系统进化关系进行了分析。鳜胃蛋白酶原基因cDNA序列全长1367 bp，5′端非翻译区43 bp，3′端非翻译区187 bp，开放阅读框(ORF)1137 bp，共编码378个氨基酸。鳜胃蛋白酶原氨基末端存在信号肽和激活肽序列，序列中含有催化活性必需的2个天冬氨酸残基和构成二硫键的6个半胱氨酸残基。鳜胃蛋白酶原氨基酸序列与其他脊椎动物胃蛋白酶原氨基酸序列的同源性为59.9%～91.2%，表明胃蛋白酶原基因在脊椎动物的长期进化中比较保守。鳜胃蛋白酶原基因的成功克隆不仅为进一步研究该基因的时空表达奠定基础，而且为鱼类胃蛋白酶原的分子特征和进化提供了新的资料。     

大口黑鲈两种脂蛋白脂肪酶基因cDNA的克隆及表达特征分析

为促进肉食性鱼类人工配合饲料开发的理论基础研究,分析鱼类脂肪代谢的机制,实验克隆了大口黑鲈2个脂蛋白脂肪酶基因LPLtype1和LPLtype2的cDNA。序列分析表明,LPLtype1基因cDNA序列全长2 156 bp,编码516个氨基酸;LPLtype2基因cDNA序列全长1 710 bp,编码346个氨基酸。大口黑鲈LPLtype2与LPLtype1氨基酸序列之间的同源性为43.5%。系统进化分析表明,大口黑鲈LPLtype1和鳜LPL聚为一支,大口黑鲈LPLtype2和大麻哈鱼LPLtype2紧密聚为一支。预测分析发现,大口黑鲈LPLtype1和LPLtype2基因编码蛋白的活性中心位点、N-糖基化位点、二聚体形成的保守疏水残基位点、肝素结合域等主要功能域与硬骨鱼类和其他脊椎动物对比都比较保守。运用实时定量PCR方法检测脂蛋白脂肪酶mRNA的组织分布,发现LPLtype1和LPLtype2都在肝脏中表达量最高,推测这与肝脏是最主要的营养诱导性储脂部位有关。     

斑节对虾亲环素A （cyclophilinA） 基因的克隆及启动子序列分析

通过同源克隆和RACE技术获得斑节对虾(Penaeus monodon)亲环素A(cyclophilin A,CYPA)的cDNA序列.根据已克隆的cDNA序列设计引物,利用染色体步移技术(Cenomic DNA walking)从斑节对虾卵巢组织中克隆了CYPA基因的启动子和基因组DNA序列.测序结果表明,斑节对虾亲环素A(PmCYPA)cDNA全长834 bp,其中开放阅读框(open reading frame,ORF)为495 bp,可编码164个氨基酸;5'非编码区为31 bp,3'非编码区为308 bp.从斑节对虾基因组文库中扩增的CYPA基因组DNA全长3 181 bp,其中包括启动子区域1 173 bp,1个内含子426 bp,2个外显子序列:分别为101 bp和399bp.在5'UTR上游区域有明显的启动子序列,包含一个GC盒和2个CAAT盒,同时还包含AP1、CRE等调控元件,符合真核生物典型的启动子特征.分析基因的结构表明所克隆的PmCYPA符合PPlase家族成员特征,属于此基因家族成员.     

虾夷扇贝肌动蛋白基因cDNA序列克隆与分析

采用RT-PCR和RACE法从虾夷扇贝闭壳肌中分离和克隆了肌动蛋白基因cDNA全长序列.肌动蛋白基因cDNA全长1775 bp(不包括poly A),5′端非编码区94 bp,3′端非翻译区551 bp,阅读框1131 bp,编码376个氨基酸.在基因组DNA中,该基因被一个内含子分为两段,内含子位于第42和第43个氨基酸之间,长度为2041 bp.系统发育分析显示该肌动蛋白属于α类型.     

半滑舌鳎颗粒酶B基因的克隆和表达分析

颗粒酶(granzyme, Gzm) B是免疫炎症反应必不可少的介质,可激活半胱天冬酶3,进而诱导靶细胞的凋亡。本研究通过PCR扩增和RACE技术获得了半滑舌鳎(Cynoglossus semilaevis)颗粒酶B基因(CsGzmBl)的全长cDNA序列,并对其序列特征和表达水平进行了分析。结果显示,CsGzmBl cDNA全长为923 bp,开放阅读框长度为780 bp,编码259个氨基酸(前19个氨基酸残基为信号肽序列),5′非编码区为49 bp,3′非编码区为94 bp。CsGzmBl的基因组结构比较保守,由5个外显子和4个内含子组成。CsGzmBl蛋白包含2个N端糖基化位点、1个催化三联体“His63Asp112Ser207”、1个“PHSRPYMA”结构域及6个保守的半胱氨酸。荧光定量PCR结果显示,CsGzmBl在半滑舌鳎健康成鱼不同组织中均有表达,其中,在脾脏中表达量最高,头肾、中肾、肝脏和鳃中表达量次之,在肌肉中表达量最低。与对照组0 h相比,CsGzmBl在哈维氏弧菌(Vibrio harveyi)感染后的不同时间点的脾脏、肠、肝脏、皮肤、鳃和肾脏中的表达水平均有不同程度的上调。研究表明,CsGzmBl基因在半滑舌鳎抵御哈维氏弧菌感染过程中发挥重要作用。     

圆斑星鲽促性腺激素释放激素基因克隆及表达特性

采用RACE技术,在圆斑星鲽(Verasper variegatus)脑中克隆到3种GnRH基因的cDNA序列: cGnRH-Ⅱ、sGnRH和sbGnRH.每种GnRH都包括1个信号肽、1个Gly-Lys-Arg连接序列和1个GnRH相关肽.其中, cGnRH-Ⅱ的cDNA全长为568 bp,编码85个氨基酸, ORF为255 bp,5′UTR为141 bp,3′UTR为169 bp.sGnRH的cDNA全长为457 bp,编码90个氨基酸, ORF为270 bp,5′UTR为41 bp,3′UTR为143 bp.sbGnRH的cDNA全长为381 bp,编码98个氨基酸, ORF为294 bp,5′UTR为48 bp,3′UTR为36 bp.分析了3种GnRH基因的编码氨基酸序列与其他脊椎动物的同源性,圆斑星鲽 GnRH 与鲽形目鱼类氨基酸同源性最高,其次为鲈形目鱼类.对3种 GnRH 基因的系统进化分析表明,圆斑星鲽 GnRH 基因与其他鲽形目鱼类亲缘关系最近,其次为鲈形目、鲑形目和鳗鲡目鱼类.荧光定量PCR分析表明,3种GnRH基因都在脑中表现出最高表达水平且具有性别特异性表达模式:雌性中不同组织的 GnRH mRNA 表达水平都相应高于雄性.组织表达分析表明, cGnRH-Ⅱ的 mRNA 仅在脑中表达,而sbGnRH在各个组织都有表达, sGnRH仅在脑、垂体和性腺中表达.脑中sbGnRH mRNA的表达水平在卵巢成熟过程中变化显著(P<0.05),而其他两种GnRH的mRNA表达水平变化不显著(P>0.05).本研究首次在圆斑星鲽脑中克隆到3种GnRH基因,其组织和季节表达水平变化表明sbGnRH可能是圆斑星鲽生殖调控的关键GnRH类型,本结果可为圆斑星鲽生殖调控机制和人工繁育技术研究提供理论支撑.     

04斜带石斑鱼干扰素调节因子 1 基因的克隆及其原核表达

对斜带石斑鱼(Epinephelus coioides)的干扰素调节因子1(Interferon regulatory factor 1,IRF-1)基因进行了克隆、测序及原核表达.以PBS做对照,利用polvI:C刺激斜带石斑鱼,然后取其肝脏、头肾、脾脏提取总RNA,随机引物反转录获得cDNA.根据相近物种IRF-1的保守序列设计引物,通过PCR得到了保守片段,通过RACE-PCR获得了斜带石斑鱼IRF-1基因的全长cDNA.基因全长为1 730 hp,完整开放阅读框(ORF)906 bp,编码302个氨基酸,5'非编码区153 bp,3'非编码区671 bp.将斜带石斑鱼ORF与其他物种进行比对发现,与海鳜(Siniperca chuatsi)同源性最高,为85%.将根据斜带石斑鱼IRF-1 ORF推测的氨基酸序列与其他物种进行比对,发现斜带石斑鱼与海鳜同源性为90%,与金头鲷(Sparus aurat)为88%,与乌鳢(Channa argus)为86%,与大菱鲆(Scophthalmus maximus)为82%.利用ORF序列,构建原核重组表达质粒.重组质粒经PCR、酶切鉴定及测序验证后转化大肠杆菌BL21(DE3),经IPTG诱导,重组蛋白成功表达,表达蛋白的分子量约55 kD.诱导温度为30℃时,重组蛋白以可溶性蛋白和包涵体2种形式存在.     

圆斑星鲽MHC IIB基因结构、多态性及组织表达分析

通过表达序列标签法和cDNA末端快速扩增(RACE)技术,分离和克隆了圆斑星鲽(Verasper variegatus)主要组织相容性复合体(MHC)IIB的全长cDNA序列,该cDNA全长为1144bp,5'UTR(untranslated region)为7bp,3'UTR为450bp,开放阅读框(ORF)长度为687bp,可编码228个氨基酸,包含信号肽、抗原结合域(β1)、IGC区(β2)、跨膜区和胞质区5个结构域。同源分析表明,圆斑星鲽MHCIIB氨基酸序列与其他硬骨鱼具有49%～79%的同源性,与鼠、人、红原鸡和护士鲨的相似性较低,分别为34%、33%、31%和30%。圆斑星鲽MHC ⅡB基因含有5个内含子,与其他硬骨鱼不同,其β2结构域编码区内存在1个109bp的内含子。根据获得的MHC ⅡB基因组序列设计特异性引物,在10尾野生圆斑星鲽中扩增了包括完整内含子1和外显子2的长度约388bp的DNA片段,PCR产物直接测序后发现在270bp的抗原结合域中共有23个位点发生变异,密码子第1位和第2位的变异明显高于第3位。利用荧光定量PCR分析组织表达发现,MHCIIB基因在健康圆斑星鲽9种组织中均有...     

剑尾鱼卵黄蛋白原C(Vg C)全长cDNA的克隆及表达

利用逆转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)等方法,克隆获得剑尾鱼卵黄蛋白原C(Vg C)基因的全长cDNA序列。剑尾鱼Vg C基因cDNA序列全长4 011 bp,其5′非编码区包含12 bp和3′非编码区包含246 bp;含有一个3 753 bp的开放阅读框(ORF),编码1 250个氨基酸,推测其编码氨基酸分子量大小为141.7 ku,编码氨基酸序列与其他鱼类卵黄白原C编码氨基酸序列相似性在44%～85%。荧光定量PCR结果显示,Vg C在剑尾鱼肝脏中表达量最高,脾、肾、卵巢中有微量表达,脑、肌肉、鳃中几乎没有检测到表达;对不同时间暴露在雌激素中剑尾鱼肝脏进行实时荧光定量PCR表达分析的结果表明,Vg C在剑尾鱼肝脏中第5天表达量最高,随后降低,第9天后维持相对较低的表达量。研究首次克隆了剑尾鱼Vg C基因全长cDNA序列,并对Vg C在剑尾鱼体内表达组织器官分布及雌激素诱导后不同时间表达谱进行了初步研究,为剑尾鱼生殖生理及环境污染物监测应用等不同领域研究打下重要基础。     

Neutral lipase and phospholipase activities in scallop juveniles (Pecten maximus) and dietary algae

Activities of the digestive enzymes neutral lipase and phospholipase A₂ were present in 2‐mm scallop juveniles, indicating an endogenous ability to digest lipids. When determined in the dietary microalgae, lipolytic activities of both enzymes differed significantly, being highest in Isochrysis galbana and Tetraselmis sp. and lowest in Chaetoceros mülleri. Compared with the activities found in scallop juveniles, enzyme activities from the microalgae were rather low and did not seem to contribute considerably to their digestive process. However, ingested algae seemed to have a regulating effect on the phospholipase activity in scallops, as juveniles fed I. galbana had a significantly lower activity than juveniles of the other treatments. No such effect was evident for neutral lipase, indicating different regulation mechanisms for both enzymes. Dietary effects on digestive enzymes in scallop juveniles should be considered when choosing new potential algal species as feed or for future formulation of artificial diets.     

Cloning of lipoprotein lipase (LPL) and the effects of dietary lipid levels on LPL expression in GIFT tilapia (Oreochromis niloticus)

Genetically improved farmed tilapia (GIFT) (Oreochromis niloticus) is an important aquaculture species. Lipoprotein lipase (LPL) is considered as a key enzyme in lipid metabolism and deposition. The present study was conducted to investigate the nutritional regulation of LPL in GIFT. We cloned and characterized the LPL gene from GIFT. Finally, we determined the effects of dietary lipid levels and refeeding on hepatic LPL gene expression in GIFT. The LPL gene of GIFT (Oreochromis niloticus) (O.nLPL) was 2,298 bp in length and encoded 515 amino acids. Sequence analysis showed that O.nLPL shared 57.3–87.9 % identity with LPLs from other piscine species. To study LPL expression patterns, juveniles GIFT were fed diets containing 3.7, 7.7 or 16.6 % crude lipid for 90 days and the expression of hepatic O.nLPL was examined using real-time PCR. The abundance of hepatic LPL mRNA increased with increasing dietary lipid. The expression of O.nLPL mRNA in the 16.6 % dietary lipid group was significantly higher than that of the 3.7 % lipid group (P < 0.05). The expression of O.nLPL was increased in GIFT following a 48-h fast and decreased 12 h after refeeding. Hepatic LPL mRNA returned to fasting levels 48 h after refeeding. In summary, high dietary lipid induced expression of liver O.nLPL, and expression of liver LPL is regulated by fasting and refeeding.     

The determination of lipase and phospholipase activities in gut contents of turbot (Scophthalmus maximus) by fluorescence-based assays

To study the potential of fluorescence based assays in the study of lipid digestion in fish, acyl esters of 4-methylumbelliferone and 1-acyl-2-[6 (7 nitro-1,3 benzoxadiazol-4-yl)amino]caproyl labeled phosphatidylcholine compounds (NBD-PC) were used as substrates for the assay of neutral lipase and phospholipase, respectively, in the gut contents of turbot. 4-Methylumbelliferyl hepatanoate (4-MUH) was hydrolysed at a higher rate than the butyrate or oleate esters whilst the hexanoic (C6) ester of NBD-PC was a more convenient substrate for the phospholipase assay than the dodecanoic (C12) ester. Neutral lipase activity was almost 10% higher when 50 mm potassium phosphate buffer pH 7.8 was used instead of 0.01 m citrate/sodium phosphate buffer pH 7.2. Both assays were very sensitive: neutral lipase and phospholipase activities were detectable at a minimum protein concentration in the digesta of 0.04 and 1.25 mg/ml, respectively. When the variations in lipolytic activities with gut segment and with size of fish were examined neutral lipase activity was always found to be higher in the hindgut and rectum segments than in the foregut. Although phospholipase activity was also found to be highest in the hindgut of the largest fish examined (av. wt. 182.3g), in fish of average weight 8g fish the activity was similar in all three segments. In the digesta from the whole gut of smaller fish (av. wt. 0.2, 0.6 and 1.43g) neutral lipase and phospholipase activities increased with increasing body mass when expressed as per ml of digesta. It is concluded that fluorescence-based assays are applicable to the study of lipid digestion in fish of different size.     

Cloning and characterization of lipoprotein lipase (LPL) and the effects of dietary lipid levels on the expression of LPL in the redlip mullet (Liza haematocheila)

The genetic improvement of redlip mullet Liza haematocheila through breeding programmes is of interest for this important aquaculture species. Lipoprotein lipase (LPL) is a key enzyme in lipid deposition and metabolism. The purpose of this study was to investigate the nutritional regulation of LPL in redlip mullet. We cloned and identified the LPL gene, determined LPL gene expression in various tissues, and examined the effect of dietary lipid level on hepatic LPL gene expression. The LPL gene of redlip mullet Liza haematocheila (L.hLPL) was 2,395 bp in length and encoded 516 amino acids. Sequence analysis showed that L.hLPL shared 61%–90.3% identity with LPLs in other species. Expression patterns of hepatic L.hLPL were studied in redlip mullet fed diets containing 2.0, 4.8, 7.5, 9.8, 12.0 or 14.6 g/kg, crude fat for 60 days by real‐time quantitative PCR. The abundance of LPL mRNA in hepatic tissue increased with the increase in dietary fat. The expression L.hLPL mRNA was significantly higher in the groups fed diets with 14.6 and 12.0 g/kg fat than in the other groups (p < .05). Gene expression was significantly higher in the abdominal fat of redlip mullet (p < .05) compared with other tissues. In conclusion, a high fat diet (9.79–14.59 g/kg) induces L.hLPL expression in abdominal fat.     

不同饲料对银鲳幼鱼增重率、肝脏脂酶及抗氧化酶活性的影响

利用营养学与生物化学的方法,研究分析了不同饲料对银鲳幼鱼增重率、饲料系数、肝脏脂酶及抗氧化酶活性的影响。试验共设4组不同饲料,依次为饲料1(新鲜鱼肉糜)、饲料2(新鲜鱼肉糜+饲料)、饲料3(新鲜鱼肉糜+饲料+蛏子肉糜)和饲料4(新鲜鱼肉糜+饲料+蛏子肉糜+桡足类)。试验用银鲳幼鱼的平均体重为(4.80±0.11)g,每组饲料设3重复,试验周期为9周。研究结果显示,不同饲料可显著影响银鲳的增重率,各饲料组中以饲料1组的增重率最低,并显著低于其它各饲料组;饲料4组的增重率最高,并显著高于饲料2、3组的增重率(P<0.05)。饲料系数以饲料1组最高,且显著高于其它3组饲料组;饲料2、3、4组饲料系数间并无显著性差异(P>0.05)。饲料1组肝脂酶与总脂酶活性均显著低于其它3组饲料组(P<0.05),但饲料2、3、4组间肝脏脂蛋白脂酶、肝脂酶和总脂酶活性均未有显著性差异(P>0.05)。各组间肝脏超氧化物歧化酶(SOD)与谷胱甘肽过氧化物酶(GSH-PX)活性均未有显著性差异(P>0.05),但均以饲料1组最低、饲料4组最高;饲料1、2、3组间肝脏过氧化氢酶(CAT)活性未有显著性差异(P>0.05),但饲料4组CAT活性显著高于饲料1组的CAT活性(P<0.05)。分析结果表明,单纯利用鱼肉糜作为饲料不利于银鲳的生长;饲料中添加桡足类不但有助于促进银鲳的生长,也可提高其抗氧化能力。     

Growth performance,nutrient digestibility and lipase activity in juvenile rainbow trout (Oncorhynchus mykiss) fed fat powder in diet containing emulsifiers (cholic acid and Tween‐80)

This research was conducted to investigate the effects of two dietary emulsifiers on nutrient digestibility and lipase activity in rainbow trout (Onchorhyncus mykiss). A basal rainbow trout diet containing fat powder supplemented with 10 and 5 g kg^?1 of cholic acid, and 20 and 40 g kg^?1 of Tween‐80. Control diet contained no emulsifiers with fat powder was replaced by fish oil. Each diet was randomly assigned to 1,500‐L tanks in triplicate. Juvenile rainbow trout with an initial weight of 27.32 ± 2.03 g were randomly distributed in the experimental tanks. The results showed that growth parameters did not change by the addition of the two emulsifiers (p > .05). Total triglyceride content was significantly higher in control fish fed diet containing fish oil (p < .05), while serum cholesterol content showed no significant differences among treatments (p > .05). Control diet resulted in a higher fat digestibility than those of other experimental diets. However, protein and ash digestibilities in diet containing emulsifier were higher than those of the control diet. Control group showed the lowest lipase activity, whereas 20 g kg^?1 Tween‐80 diet caused the highest lipase activity among treatments (p < .05). In conclusion, it seems that a higher lipase activity induced by the emulsifiers could not compensate for the negative impacts of fat powder on the experimental diets.     

Dietary n‐3 LC‐PUFAs affect lipoprotein lipase (LPL) and fatty acid synthase (FAS) activities and mRNA expression during vitellogenesis and ovarian fatty acid composition of female silver pomfret (Pampus argenteus) broodstock

We studied the effects of dietary n‐3 LC‐PUFAs on the activities and mRNA expression levels of tissue lipoprotein lipase (LPL) and fatty acid synthase (FAS) during vitellogenesis and ovarian fatty acid composition in female silver pomfret broodstock. Broodstock were fed one of four experimental diets for 185 days: FO (100% fish oil), FSO (70% fish oil + 30% soybean oil), SFO (30% fish oil + 70% soybean oil) or SO (100% soybean oil). The results revealed that hepatic LPL and FAS and ovarian FAS activities and mRNA expression levels significantly increased at vitellogenesis and postvitellogenesis relative to previtellogenesis, with no significant differences between these two stages, except for hepatic LPL mRNA expression. Dietary n‐3 LC‐PUFAs decreased tissue FAS and increased LPL activities and mRNA expression levels. The ovarian concentrations of 20:4n‐6 (ARA), 20:5n‐3 (EPA), 22:6n‐3 (DHA) and n‐3 LC‐PUFAs were directly influenced by n‐3 LC‐PUFA levels. Total n‐3 LC‐PUFA concentrations in SO were 57% lower than those in FO, while 18:2n‐6 concentrations in SO were 4.7 ×  higher than those in FO. These results revealed that high dietary n‐3 LC‐PUFAs levels significantly affected tissue lipid metabolism in female silver pomfret broodstock during vitellogenesis by upregulating LPL and downregulating FAS.     

脂肪酶对南方鲇生长性能、消化酶活性及血液生化指标的影响

在基础饲料中分别添加0、0.1、0.3 g/kg的脂肪酶,饲喂平均体重(57.6±0.02)g的南方鲇(Silurus meridionalis Chen)60 d,测定南方鲇的生长性能、饲料表观消化率、消化酶活性及血液生化指标。结果显示:添加0.1 g/kg和0.3 g/kg的脂肪酶,增重率比对照组分别提高了4.0%和5.8%,但差异不显著(P>0.05),饲料系数分别下降了5.0%和6.6%,差异显著(P<0.05),肥满度和粗脂肪消化率显著提高(P<0.05),对内脏指数、肝脏指数、肠脂指数、胰蛋白酶活力、肠蛋白酶活力、肠脂肪酶活力及血液生化指标均没有显著影响(P>0.05)。添加0.3 g/kg的脂肪酶显著提高胰脂肪酶活力(P<0.05),添加0.1 g/kg的脂肪酶对胰脂肪酶活力没有显著影响(P>0.05)。结果表明,在脂肪水平为10%的饲料中添加0.3 g/kg的脂肪酶可以提高胰脂肪酶活力,提高脂肪消化率,改善饲料利用率。     

Effect of dietary fish meal and fish oil replacement on lipogenic and lipoprotein lipase activities and plasma insulin in gilthead sea bream (Sparus aurata)

The effects of a double replacement of fish oil (FO) and fish meal (FM) by dietary vegetable ingredients in juvenile gilthead sea bream (Sparus aurata L. 1758) on some indices of lipid metabolism and plasma insulin levels were analysed. Four experimental diets with a replacement of 75% of FM by plant proteins (PP) were administered. Added oil was either FO (75PP/FO diet), or a vegetable oil mix (VO), replacing 33%, 66% or 100% of FO (75PP/33VO, 75PP/66VO, 75PP/100VO diets). Another diet with 50% of substitution of FM by PP and with 100% of VO was also tested (50PP/100VO diet). Final body weight was similar in all diet groups, except for the 75PP/100VO group, which presented lower values. Circulating insulin levels increased with feed administration in all groups and no differences between diets were observed, with the exception of the 75PP/FO group, which presented higher plasma insulin values. In adipose tissue, glucose‐6‐phosphate dehydrogenase and malic enzyme activities decreased with the inclusion of vegetable oil, especially 5 h after feeding. Diet had no significant effect on the hepatic activity of either enzyme. Lipoprotein lipase activity decreased in white muscle and adipose tissue with the replacement of fish oil in 75PP diets, 5 h after feeding. In conclusion, the use of a combined replacement of fish oil and fish meal by vegetable ingredients in gilthead sea bream permits satisfactory growth, with moderate changes in tissue lipogenesis and lipid uptake.