太平洋鳕神经坏死病毒衣壳蛋白(CP)的原核表达及条件优化

为深入研究太平洋鳕Gadus macrocephalus神经坏死病毒(Pacific cod nervous necrosis virus,PCNNV)衣壳蛋白(capsid protein,CP)的功能,将PCNNV cp基因连接到p ET-32a原核表达载体中,构建原核表达重组载体p EVCP,将其转化至大肠杆菌E.coli BL21(DE3)表达系统中进行融合表达,对诱导剂IPTG浓度、诱导时间和温度等诱导表达条件进行优化,采用SDS-PAGE及质谱技术鉴定表达产物。结果表明:本研究中成功构建了重组载体p EVCP,表达的目的蛋白相对分子质量约为55 000;目的蛋白经质谱鉴定,有6个肽段的氨基酸序列与预期一致;对重组菌株p EVCP/BL21的最佳诱导条件为IPTG浓度0.1 mmol/L、最佳诱导时间3 h、温度30℃。本研究结果可为后续冷水鱼类病毒疫苗的研制及病毒快速检测试剂盒的研发奠定基础。     

Effect of dietary supplemental l‐carnitine on growth performance,body composition and antioxidant status in juvenile black sea bream,Sparus macrocephalus

Effects of dietary l ‐carnitine were studied in juvenile black sea bream (Sparus macrocephalus). The semipurified basal diet [crude protein 450 g kg^?1 dry matter (DM); crude lipid 126 g kg^?1 DM] was formulated to choose white fishmeal as the protein source and fish oil plus corn oil (1 : 1) as the lipid source. Six diets (control + diets 1–5) containing 0.1, 0.12, 0.16, 0.24, 0.39 and 1.1 g of l ‐carnitine kg^?1 diet were fed to triplicate groups of black sea bream (initial weight 13.10 ± 0.05 g) for 8 weeks. At the end of the feeding trial, growth performance, body composition and antioxidant status were determined. The results showed that relative growth rate (RGR) was significantly improved by the elevation of dietary l ‐carnitine level from 0.1 to 0.24 g kg^?1, but decreased with further increment (P < 0.05). Lipid content decreased significantly (P < 0.05) in the dorsal muscle whereas increased (P < 0.05) in the liver with the addition of dietary l ‐carnitine. Dietary l ‐carnitine supplements elevated enzymatic antioxidants (superoxide dismutase, SOD; catalase, CAT; glutathione‐S‐transferase, GST) activities (P < 0.05) yet decreased the content of non‐enzymatic factor, total sulphydryl groups (TSH) (P < 0.05). In summary, the optimum dietary l ‐carnitine level was 0.284 g kg^?1 diet by second‐polynomial regression analysis based on RGR (y = ?647.4x² +367.97x + 234.55; R² = 0.977, x = dietary l ‐carnitine levels, y = RGR), and dietary l ‐carnitine addition within the levels adopted in our study could depress lipid peroxidation in tissues of juvenile black sea bream.     

Australian sperm whales from different whaling stocks belong to the same population

1. Understanding the factors driving population structure in marine mammals is needed to evaluate the impacts of previous exploitation, current anthropogenic threats, conservation status, and success of population recovery efforts.
2. Sperm whales are characterized by a worldwide distribution, low genetic diversity, complex patterns of social and genetic structure that differ significantly within and between ocean basins, and a long history of being commercially whaled. In Australia, sperm whales from the (International Whaling Commission assigned) southern hemisphere ‘Division 5’ stock were very heavily exploited by whaling.
3. The present study assessed the potential effects of whaling on the genetic diversity of sperm whales in Australia and the population genetic structure of these whales within a global context. A combination of historical and contemporary sperm whale samples (n = 157) were analysed across six regions, from south-eastern Australia (‘Division 6’ stock in the Pacific Ocean) to south-western Australia (‘Division 5’ stock in the Indian Ocean).
4. Sperm whales sampled from the ‘Division 5’ and ‘Division 6’ stocks belong to the same population based on nuclear and mitochondrial DNA (mtDNA) analyses. Four novel sperm whale mtDNA haplotypes were identified in animals from Australian waters. Levels of genetic diversity were low in Australian sperm whales but were similar to those previously reported for populations in the Indian and Pacific Oceans.
5. Given the genetic distinctiveness of sperm whales in Australian waters from other regions in the Pacific and Indian Oceans, and the lack of recovery in population numbers, further scientific studies are needed to increase our understanding of population dynamics and the effectiveness of threat management strategies in this species.

     

Expression of four muscle proteins at different growth stages of Günther's walking catfish Clarias macrocephalus

The complete cDNA sequences of four contractile muscle genes of walking catfish Clarias macrocephalus were characterized by assembling partial EST sequences from a skeletal muscle cDNA library. The four genes were parvalbumin 4 (PV4) (670 bp), troponin C (TnC) (1065 bp), troponin I (TnI) (843 bp) and myosin light chain 3 (MLC3) (953 bp), leading to deduced amino acid sequences of 109, 160, 176 and 150 residues respectively. During the larval stage, TnC mRNA showed the highest levels of expression with a 1.4‐fold increase from day 1 to day 30 post hatch. At 90 days, the relative expression levels of PV4, TnC and MLC3 were the highest, with similar proportions in the skeletal muscle, corresponding to the highest relative growth rate of walking catfish. Expression of the three calcium‐binding proteins remained high in 6‐month‐old fish, with higher levels of PV4. The different proportions of muscle proteins expressed suggested the significance of their contributions to fish growth and appeared to be correlated with the functional properties of muscle cells, which were observed from changes in the swimming activity of the fish.     

不同光照强度对太平洋鳕仔鱼摄食的影响

研究不同光照强度对太平洋鳕仔鱼摄食的影响。在0～1 000 lx光照范围内,对不同光照强度、摄食不同时间太平洋鳕仔鱼摄食量进行了测定,结果表明:仔鱼500 lx时摄食强度最大,其次为700、300 lx,摄食率在500 lx时最高,并在摄食开始后10 min时达到最大,并随着时间的延长而降低。     

真鲷与黑鲷杂交繁育技术研究

通过对真鲷黑鲷杂交繁育,培育出一种能结合父母本性状,具有生长速度快、抗逆性强的杂交子一代。2006年3月从海区选购真鲷（Pagrosomusmajor）、黑鲷（Spraus macrocephalus）亲鱼,通过强化培育,性腺促熟,于4月进行干法人工授精。通过对真鲷♀&#215;黑鲷8及黑鲷♀&#215;真鲷♂获得杂交受精卵,结果真鲷♀&#215;黑鲷♂受精率为87．4％,孵化率为85％,出苗率14．28％;黑鲷♀&#215;真鲷♂受精率为92．1％,孵化率为90％,出苗率7．2％,该批次共获得正交反交杂交鱼苗32000尾。从杂交实际结果看,不管正交还是反交都比自交的结果差,说明它们之间存在生殖隔离,杂交繁育难度相当大,要达到规模生产。需要继续研究、试验。     

人工鱼礁竖板不同方形孔径对黑鲷幼鱼诱集效果研究

建设人工鱼礁是修复海洋生境、养护渔业资源的重要措施.钢筋混凝土鱼礁是常用的人工鱼礁类型,鱼礁竖板(侧板、内部间板)方形开孔是钢筋混凝土鱼礁结构设计中主要设计参数,竖板方形孔径大小是影响鱼类诱集效果的重要因素.以人工鱼礁区常见的礁栖鱼类黑鲷(Sparus macrocephalus)幼鱼为研究对象,设计制作具有不同方形孔...     

摄食水平和饵料种类对3种海洋鱼类生长和生长效率的影响

采用室内流水模拟法测定了摄食水平与铒料生物种类对真鲷、黑鲷、黑Ｊｕｅ特定生长率和生长效率的影响。３种实验鱼类的物壹生长率均随摄食水平增大呈减速增长趋势。二者间的定量关系和对数曲线描述;而其生长效率则均随摄食水平增大呈倒“Ｕ”形变化趋势。二者间的定量关系可用二次曲线描述,曲线相关检验结果表明,其特定生长率和生长效率与摄食水平之间均呈显著相关关系。依据上述定量关系,分别求得３种鱼的维持摄食量和最佳摄食     

太平洋鳕性腺发育及营养来源的初步研究

通过对大连海域太平洋鳕Gadus macrocephalus的繁殖力、卵径分布、性腺指数(GSI)和肝指数(HSI)进行周年调查,确定出太平洋鳕的繁殖期,分别测定了不同发育期太平洋鳕性腺、肝脏和肌肉的生化组成、能量密度变化,并观察了不同发育期太平洋鳕性腺和肝脏的组织学变化.结果表明:大连海域太平洋鳕的卵巢从12月开始进入第Ⅳ期,至翌年2月份排放;精巢处于Ⅳ、Ⅴ期的时间较长,11月份就有达到成熟的个体;从太平洋鳕肝脏组织学的变化来看,脂滴的体积变化很大,说明在繁殖过程中,肝脏经历了一个脂肪先积累又被大量消耗的过程;太平洋鳕在繁殖过程中,各个组织的能量密度变化较小,但粗蛋白、粗脂肪、粗灰分等成分变化较大.研究表明,太平洋鳕性腺发育的脂肪主要可能来源于肝脏,而肌肉蛋白的下降可能充当了性腺发育中蛋白质的主要来源.     

Effects of ontogeny, temperature, and light on vertical movements of larval Pacific cod (Gadus macrocephalus)

The role of behavior, especially vertical migration, is recognized as a critical component of realistic models of larval fish dispersion. Unfortunately, our understanding of these behaviors lags well behind our ability to construct three-dimensional flow-field models. Previous field studies of vertical behavior of larval Pacific cod ( Gadus macrocephalus ) were limited to small, preflexion stages (≤11 mm SL) in a narrow range of thermal conditions. To develop a more complete picture of larval behavior, we examined the effects of ontogeny, temperature, and light on vertical responses of larval Pacific cod in experimental columns. While eggs were strictly demersal, yolk-sac larvae displayed a strong surface orientation as early as 1 day post hatch (∼ 5 mm SL). Consistent with field observations, small preflexion larvae (<10 mm SL) showed no response to varying light levels. However, there was a direct effect of temperature on larval behavior: Pacific cod larvae exhibited a stronger surface orientation at 4°C than at 8°C. The behavior of larger, postflexion larvae (>15 mm SL) in experimental columns was consistent with a diel vertical migration and independent of water temperature: fish were more widely distributed in the column, and median positions were consistently deeper at higher light levels. These laboratory observations are combined with observations from discrete-depth (MOCNESS) sampling in the Gulf of Alaska to characterize the vertical distribution of larval Pacific cod and contrast ontogenetic patterns with walleye pollock ( Theragra chalcogramma ). The vertical movements of larval Pacific cod described here will be applied in the development of dispersal projections from Gulf of Alaska spawning grounds.