Molecular cloning,characterization and expression analysis of glucose transporters from Rachycentron canadum

It is very necessary to clarify the mechanism of carbohydrate metabolism in fish due to their poor ability to utilize carbohydrates. Glucose transporters (GLUTs) are important carriers involved in glucose transport across the plasma membrane, which is the first rate‐limiting step of carbohydrate metabolism. In this study, five glucose transporters (RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9) were obtained from Rachycentron canadum. The complete cDNAs open reading frames (ORF) of RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9 were 1,476 bp, 1,530 bp, 1,551 bp, 1,539 bp and 1,578 bp (GenBank accession no. MK560257 , MK560258 , MH184460 , MH184461 and MH184462 ), encoding 491, 509, 516, 512 and 525 amino acid (aa) residues respectively. All five RcGLUTs contained a Sugar_tr domain, which is an important feature of the GLUT gene family. The multiple sequence alignment and phylogenetic relationship analyses showed that RcGLUTs were highly conserved and homologous from fish to mammals. Examination of tissue distribution showed that RcGLUTs were expressed constitutively in R. canadum. RcGLUT1‐5 and RcGLUT9 had the highest expression in the foregut, kidneys, haemocytes, muscles, liver and heart respectively. All six RcGLUTs first increased to peak levels of expression and then reduced both in the liver and muscle after fasting. The exception is that the expression levels of RcGLUT4 and RcGLUT5 in muscle were significantly lower than the control. In response to hypoxia, only RcGLUT2 in the liver and muscle and RcGLUT9 in muscle were expressed at a significantly lower level relative to the control. In sum, all RcGLUTs in the liver and muscles showed significant changes in response to fasting and hypoxia, suggesting that GLUT genes may play a role in the response to common physiological stresses.     

饲料n-3/n-6脂肪酸比值对军曹鱼生长及鱼体组织脂肪酸组成的影响

以精制鲽鱼油和豆油分别作为n-3及n-6系列脂肪酸源,配制5种不同n-3/n-6脂肪酸比值的饲料,进行连续8周饲喂军曹鱼幼鱼试验。结果表明,当饲料中的n-3/n-6脂肪酸比值为2.57时,军曹鱼的增重率和饲料效率最高;饲料中不同n-3/n-6脂肪酸比值对军曹鱼血清5项生理指标的影响不显著,但随着n-3/n-6脂肪酸比值的降低,血清中的谷丙转氨酶、谷草转氨酶、胆固醇含量表现出下降趋势;饲料中n-3、n-6多不饱和脂肪酸含量,与军曹鱼肌肉和肝脏的n-3、n-6多不饱和脂肪酸含量呈正相关。统计分析表明,军曹鱼饲料中适宜的n-3、n-6脂肪酸比例为2.98∶1。     

军曹鱼池塘人工育苗技术的初步研究

在池塘进行军曹鱼Rachycentron canadum人工育苗研究,通过解剖消化肠道以确定仔鱼开口的生物饵料种类。结果显示,个体较大的桡足类无节幼体比轮虫更适合作为军曹鱼仔鱼的开口饵料;对体长和体重进行测定,军曹鱼在鱼塘进行人工育苗,鱼苗生长迅速,7日龄的仔鱼体长达1cm,30d的鱼苗可长至15cm。     

军曹鱼幼鱼日粮锌最适添加量的研究

在以卵白蛋白为蛋白源的基础饲料（Zn含量为5．65mg／kg）中添加10、20、30、40、50mg／kg以ZnSO4为Zn源的Zn，使饲料中总Zn含量为15．97、25．66、33．90、45．85、55．88mg／kg。投喂军曹鱼幼鱼8周。试验结果表明，基础饲料组鱼出现生长不良、厌食、皮肤糜烂和白内障等症状，其成活率、增重率、血清碱性磷酸酶活性（AKP）显著低于添加Zn组（P〈0．05），骨骼、肌肉、血清和肝脏Zn含量也表现出相同趋势。10mg／kg和20mg／kgZn添加量组鱼仍出现缺Zn症状，各项测定指标随Zn添加量的增加而增大。当饲料中Zn添加量达到30mg／kg时，各项指标趋向稳定，与添加量40mg／kg和50mg／kg组无显著差异。以增重率和骨骼Zn含量为主要评价指标，通过回归分析表明，以ZnSO4为Zn源时，军曹鱼幼鱼日粮Zn的最适添加量为42．86mg／kg。     

军曹鱼对营养物质的需求

军曹鱼(Rachycentron canadum)俗称海鲡,又称海龙鱼、海竺鱼、竹五、海于草、锡腊白等,属鲈形目、鲈亚目、军曹鱼科、军曹鱼属.军曹鱼是近年来发展迅速的一种近岸网箱养殖鱼类,广泛分布于除东太平洋以外的热带亚热带等水温较高的海域[1].我国沿海亦有分布,但产量较低.     

Effects of dietary manganese sources and levels on growth performance,relative manganese bioavailability,antioxidant activities and tissue mineral content of juvenile cobia (Rachycentron canadum L)

This study was conducted to compare the effects of manganese sulphate (Mn‐S), glycine manganese(Mn‐Gly) and manganese 2‐hydroxy‐4‐(methylthio)butyrate (Mn‐HMB) on juvenile cobia, Rachycentron canadum L. Treatments consisted of 0, 2, 4, 8, 16 or 32 mg supplemental Mn kg^?1 from Mn‐S, Mn‐Gly or Mn‐MHB. Growth performance, manganese status, antioxidant activities and tissue mineral content were analysed after a 70‐day feeding period. Specific growth rate (SGR) increased with feeding 6.29 to 12.65 mg Mn kg^?1 diet from the Mn‐S or 6.86 to 12.39 mg Mn kg^?1 from the Mn‐Gly or 6.50 to 8.33 mg Mn kg^?1 from the Mn‐HMB and then plateaued above these levels. Feed conversion ratio (FCR) show decreasing first and then increased trend. Survival rate (SR) were not affected by the dietary treatments (P > 0.05). Fish fed diets supplemented with manganese at levels of 4–32 mg Mn kg^?1 had obviously higher hepatic Mn‐SOD activity (P < 0.05); on the contrary, hepatic has lower malondialdehyde (MDA) content (P < 0.05) than fish fed the basal diet. The manganese concentrations of whole body and vertebrae increased with increasing dietary Mn levels from 2–32 mg Mn kg^?1 (independent on manganese sources). Dietary Mn supplementation did not significantly influence the copper concentrations of whole body and vertebrae, the zinc concentrations of whole body and liver. Analysis by the broken‐line regression of SGR indicated that the optimal dietary Mn requirements in juvenile cobia were 15.42, 11.22 and 10.50 mg Mn kg^?1 diet from Mn‐S, Mn‐Gly or Mn‐HMB respectively.     

Resistance of Cobia,Rachycentron canadum,Juveniles to Low Salinity,Low Temperature,and High Environmental Nitrite Concentrations

Abstract

Resistance of juvenile cobia, Rachycentron canadum, to low salinity, low temperature and high nitrite concentrations was examined under laboratory conditions. After acclimating juveniles to a salinity of 20 g/L (27.3°C), salinity was decreased by 2 g/L/day. The first fish died at a salinity of 8 g/L and 80% offish were dead within 24 hours of exposure to 2 g/L. Acclimation offish to 22.6°C (21 g/L salinity) followed by a temperature reduction of 0.53°C/day resulted in initial mortality at 12.9°C. The median-lethal temperature was 12.1°C and all fish were dead by the time the temperature reached 10.4°C. Fish exposed for 96 hours to nominal ? 32 mg/L nitrite-N survived. Results of this study indicate that cobia juveniles require a salinity and temperature of > 8.0 g/L and > 12.9°C, respectively, and that environmental nitrite should not be deleterious at concentrations normally found in aquaculture systems.     

温度和体质量对军曹鱼生长及氮收支的影响

为调查军曹鱼生长和氮收支与温度和体质量的关系,采用鱼类生物能量学的原理和方法,开展了不同温度(21、27和33 ℃)和体质量(10、20、50和100 g)对军曹鱼生长和氮收支的影响研究,并建立了生长——温度/体质量关系及不同温度和体质量条件下的氮收支方程。结果显示:(1)温度和体质量对军曹鱼生长影响显著,且两者之间存在交互作用。军曹鱼在实验的最高水温(33 ℃)及最小体质量(10 g)时生长最快,在实验的最低水温(21 ℃)及最大体质量(100 g)时生长最慢。(2)实验的体质量范围内,军曹鱼特定生长率均随水温升高呈显著增长趋势;在实验的温度范围内,军曹鱼特定生长率随体质量增加呈减速下降趋势,两者间的关系采用幂函数定量描述。多元回归分析表明,模型LnSGR=a+bLnW+cT+dT²+eTLnW能较好地拟合军曹鱼特定生长率、温度及体质量间的关系。(3)军曹鱼摄食氮、排粪氮、排泄氮和生长氮受温度和体质量影响显著,随温度升高而增加,随体质量增加趋于减少。(4)不同温度和体质量组军曹鱼氮收支方程存在差异,且温度对氮收支方程的影响大于体质量。对同一体质量组,水温27和33 ℃时生长氮占食物氮的比例明显高于水温21 ℃时,而水温27和33 ℃时代谢氮占食物氮的比例则明显低于水温21 ℃时;对同一温度组,军曹鱼氮收支方程变幅较小。(5)在所有氮收支方程中,摄食氮用于排泄的比例大于68%。研究表明,在实验的温度和体质量范围内,环境温度偏低不利于军曹鱼生长;排泄是军曹鱼氮支出的最主要途径;水温27~33 ℃时军曹鱼氮收支方程相对恒定。     

军曹鱼全基因组微卫星特征分析与多态性标记的筛选及应用

为更好地开发军曹鱼(Rachycentron canadum)高多态性微卫星分子标记,本研究基于军曹鱼全基因组测序结果,利用MISA1.0软件对其简单重复序列(simple sequence repeat, SSR)的位点信息进行检索及分析。结果显示,基因组中共筛选出1~6个核苷酸为重复单元的SSR位点344 820个,其中,以单核苷酸重复序列最多(174 146个),占SSR总数的50.50%;其次为二核苷酸重复和三核苷酸重复序列,分别占SSR总数的30.23%和14.02%。在SSR包含的重复单元中,单核苷酸重复以A/T类型为主,AC/GT是二核苷酸的优势重复单元类型。军曹鱼基因组SSR核心序列重复次数在4~275次范围内波动,单核苷酸SSR重复数为10的最多,二核苷酸SSR重复次数为6的最多。本研究设置长度≥12 bp为筛选高多态性SSR位点的标准,共获得361 684个位点。在上述位点中随机选取100个候选位点进行基因分型检测,利用从中筛选到多态性较高的10个SSR标记分别对北海、陵水、硇洲、徐闻和三亚5个养殖群体进行遗传多样性分析,145尾个体中共检测到69个等位基因,观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)平均值分别为0.628、0.706和0.653。相关研究表明,军曹鱼基因组中SSR位点类型较为丰富、多态性潜能较高,从中筛选获得的多态性SSR标记可为军曹鱼分子标记辅助育种、群体遗传多样性评价等研究提供有力支持。     

Growth and intestinal morphology in cobia (Rachycentron canadum) fed extruded diets with two types of soybean meal partly replacing fish meal

Juvenile cobias, Rachycentron canadum, were fed extruded diets containing toasted defatted soybean meal (SBM) or untoasted defatted SBM [white flakes (WF)] to study growth and feed conversion, and to study if SBM induces morphological changes in the gastrointestinal (GI) tract. Three diets were produced: a fish meal‐based control diet (FM diet) with 558 g FM kg^?1, and two diets with 335 g FM and either 285 g SBM kg^?1 (SBM diet) or 285 g WF kg^?1 (WF diet). The diets were extruded at approximately 120°C with 280 g kg^?1 moisture. Triplicate groups of cobias (mean weight: 25.9 g) were fed the diets during 6 weeks. Feed intake of the FM and SBM diets were not significantly different, whereas the cumulative feed intake of cobias fed the WF diet was lower (P < 0.05) than that of cobias fed the FM and SBM diets after the first 21‐day period. Specific growth rate and feed conversion ratio were not significantly different between cobias fed the FM and SBM diets, but significantly poorer results were obtained in cobias fed the WF diet. No morphological differences in the GI tract could be attributed to the diets, and cobias fed soy did not develop enteritis in the distal intestine.