BSA-ISCOMs对欧洲鳗鲡免疫效应及抗细菌性病原的效果研究

为证实免疫刺激复合物(ISCOMs)的非特异免疫增强作用,将牛血清白蛋白(BSA)制备成ISCOMs(BSA-ISCOMs).将欧洲鳗鲡(Anguilla anguilla)随机分为3组,分别用BSA-ISCOMs、BSA和磷酸盐缓冲液(PBS,对照组)腹腔注射免疫.二次免疫完成后第30d采血,测血清部分非特异性免疫指标,剩余鳗鲡分别用创伤弧菌(Vibrio vulnificus)和嗜水气单胞菌(Aeromona hydrophila)攻毒.结果显示,BSA-ISCOMs组的血清溶菌酶活力显著高于BSA组和对照组(p≤0.05);超氧化物歧化酶(SOD)和补体C3活力与对照组无显著差异(p0.05).攻毒结果显示,BSA-ISCOMs组的存活率显著高于BSA组和对照组(p≤0.05),对创伤弧菌和嗜水气单胞菌攻击的相对免疫保护率分别为43.3%和51.7%.表明BSA-ISCOMs能有效增强欧洲鳗鲡对主要病原创伤弧菌和嗜水气单胞菌的抗病能力.     

基于耳石中锶和钙元素分析破解日本六角川和筑后川河口水域刀鲚的生境履历

为了解日本有明海沿岸六角川和筑后川河口水域鳀科鲚属鱼类刀鲚（Coilia nasus）的洄游特征和生境履历,利用电子探针微区分析（EPMA）技术,对采于两水域刀鲚的矢耳石进行了锶（Sr）元素含量的X射线强度面分布及耳石最长轴锶钙元素比值（Sr:Ca）的定量线分析。结果发现10尾六角川刀鲚（LJC）中的7尾以及15尾筑后川刀鲚（ZHC）耳石核心区域的Sr:Ca比值≤ 3,显示出这些个体在淡水生境出生并早期生活于淡水的履历。紧邻核心区域随后的生长阶段,Sr面分布图谱显示出了多样对应于含量水平差异的不同颜色同心环轮变化样式,Sr:Ca比值的波动亦可相应地分成1-6个差异极其显著的阶段,显示出这些个体在随后生活史阶段中极富变化的生境转换状况。另外3尾六角川刀鲚耳石核心的Sr:Ca比值较高（3-6.7）,对应于半咸水生境;但在核心区域之外,Sr:Ca比值则高低变化不一,显示出这些六角川刀鲚个体在河口半咸水生境孵化的特征。本研究表明,二维Sr元素含量面分布图谱和基于耳石最长轴的Sr:Ca比值完全可以作为重建日本有明海沿岸各河口水域刀鲚生境履历的有效工具。     

Temporal patterns of glass eel migration (Anguilla anguilla L. 1758) in relation to environmental factors in the Western Greek inland waters

Glass eel migration of the European eel Anguilla anguilla (L., 1758) in the eastern Mediterranean is poorly known despite the increasing state of anxiety for the future of the stock. In the present study, glass eel migration was investigated from October 1999 to April 2000, using experimental fishery with fyke nets, in two coastal systems along the western Greece (Ionian Sea): at the Sagiada marsh in the delta area of Kalamas River and at the mouth of Alfios River. The main period of the glass eel entrance was from December to March and the migration pattern was similar to those observed along the Atlantic coast of southwestern Europe. The variance of daily glass eel catches between the two studied sites was significantly different. The univariate and bivariate time series spectral analysis showed that glass eel short-term freshwater migration: (1) consisted of waves with periods from 5 to 40 days and (2) was correlated with environmental factors such as water temperature, atmospheric pressure, rainfall and moonlight. The observed differences between the two sites reveal the importance of the inland ecosystem characteristics.     

我国瓯江口和珠江口凤鲚洄游模式和生境利用研究

Habitat use of the tapertail anchovy(Coilia mystus Linnaeus, 1758) from the Oujiang River Estuary and the Zhujiang(Pearl) River Estuary was studied by examining the environmental signatures of Sr and Ca in otoliths using electron probe microanalysis. Individuals from the Oujiang River had higher and varied Sr:Ca ratios(expressed as(Sr:Ca)×1 000, 3.83–13.0 average) in the otolith core regions, suggesting that they were born in brackish or sea waters, and that a freshwater habitat might not be necessary for egg hatching and larval growth.While, individuals from the Zhujiang River had lower Sr:Ca ratios(0.39–2.51 average) in the core regions,suggesting a freshwater origin. After hatching, anchovies from the Zhujiang River migrate downstream to the river estuary close to brackish water. Our results demonstrated varied habitat use for spawning during stages of early life history between the two populations, and suggested that such variations are promoting diversity of life history strategies of this species.     

3个花鳗鲡地理种群的AFLP分析

应用AFLP分子标记技术首次对澳大利亚、海南岛和菲律宾3个：仲群的共60尾花鳗鲡（Anguillamarmorata）幼鱼样品进行了遗传多样性分析。4对选择性扩增引物共扩增得到180个位点,平均每对引物扩增出45个位点。澳大利亚、海南岛和菲律宾3个种群的多态位点分别为78．33％、79．44％和84．44％,Nei’s...     

日本鳗鲡UNAG蛋白的克隆及其在非结合胆红素检测中的应用

血清中非结合胆红素含量升高与新生儿脑损伤的发展相关,但目前临床上没有方法可以直接测定非结合胆红素的血清水平.从日本鳗鲡(Anguilla japonica)中成功克隆了其UNAG基因,在原核细胞中成功表达UNAG重组蛋白,并通过Ni~(2+)-NTA进行纯化得到含量为1.8 mg/cm~3的UNAG目的蛋白,银染检测说明其纯度较高.利用该基因可以与非结合胆红素结合并发光的特异性,设计了一种新型的非结合胆红素检测方法,结果表明UNAG发光值与胆红素浓度呈良好的线性关系,回归方程为Y=17.96 X+4.333 3(R=0.994 9).该法可用于胆红素浓度的测定.     

Induction of cytochrome P4501A and biliary PAH metabolites in European eel Anguilla anguilla: Seasonal, dose- and time-response variability in field and laboratory conditions

Induction of cytochrome P4501A in fish is a well-known indicator of exposure to polycyclic aromatic hydrocarbons (PAHs) and determination of PAH metabolites in bile by fixed wavelength fluorescence (FF) or synchronous fluorescence spectroscopy (SFS), has become an useful method in monitoring programs. In this work the relationship between cytochrome P4501A (EROD activity) and levels of biliary PAH metabolites was measured in the European eel Anguilla anguilla, in both field and laboratory conditions: organisms were sampled on a seasonal basis from the Orbetello lagoon (Tuscany) to characterize the natural variability of these biological parameters, while in laboratory eels were intraperitoneally injected with benzo[a]pyrene to investigate temporal and dose-dependent induction patterns. Results showed that induction of cytochrome P450 and accumulation of PAHs metabolites in bile are not necessarily correlated either in field, or in laboratory investigations; different seasonal changes were measured in natural conditions and slight variations in dose and time response patterns were also obtained in laboratory exposures.     

复合酶制剂对欧洲鳗鲡(Anguilla anguilla)生长性能、消化酶及非特异性免疫的影响

选用540尾初重为(6.38±0.26)g的欧洲鳗鲡(Anguilla anguilla),随机分为6个处理(每个处理3个重复,每个重复30尾鱼),分别投喂添加了0.05%(Diet 1)、0.10%(Diet 2)、0.15%(Diet 3)、0.20%(Diet 4)、0.25%(Diet 5)的复合酶制剂饲料,另增设基础饲料为对照组(Diet 0),研究复合酶制剂对欧鳗生长性能、消化酶活性及非特异性免疫的影响。6周投喂结果表明:随着复合酶制剂添加量的递增欧鳗增重率和特定生长率呈先增后降趋势,其中Diet 3组增重率和特定生长率分别比对照组提高了34.22%和26.36%(P0.05),肝体指数及肥满度各组之间差异不显著(P0.05);复合酶制剂降低了鱼体粗灰分含量,对鱼体含水量、粗脂肪及粗蛋白含量影响较小,均无显著差异(P0.05);相比对照组各添加组鳗鱼肠道胰蛋白酶活性显著提高(P0.05),淀粉酶活性则随酶制剂浓度的增高呈先升后降趋势,脂肪酶活性受酶制剂影响较小,组间无显著差异(P0.05);各添加组鳗鱼血清中超氧化物歧化酶(SOD)、溶菌酶(LZM)、碱性磷酸酶(AKP)活性相比对照组均有不同程度的提高,所有添加组鳗鱼血清总蛋白含量(TP)均显著高于对照组,Diet 3组(0.15%)相比对照提高了38.86%。综上所述,饲料中添加适量的复合酶制剂具有促进鳗鲡生长、提高肠道消化酶活性和非特异性免疫的功能,本试验条件下酶制剂最适添加浓度为0.15%。     

Habitat selection and quality for multiple cohorts of young-of-the-year bluefish (Pomatomus saltatrix): Comparisons between estuarine and ocean beaches in southern New Jersey

In this study, seasonal and annual variability in the use of estuarine and ocean beaches by young-of-the-year bluefish, Pomatomus saltatrix, was evaluated by indices of abundance in coastal areas of southern New Jersey (1998–2000). Biological and physical factors measured at specific sites were correlated with bluefish abundance to determine the mechanisms underlying habitat selection. In addition, integrative and discrete indicators of bluefish growth were used to examine spatio-temporal dynamics in habitat quality and its effect on habitat selection by multiple cohorts of bluefish. Intra-annual recruitment to coastal areas of southern New Jersey was episodic, and resulted from the ingress of spring-spawned bluefish (hatch-date April) to estuarine beaches in late May to early June, followed by the recruitment of summer-spawned fish (hatch-date early July) to ocean beaches from July to October. Bluefish utilized estuarine and ocean beaches in a facultative manner that was responsive to dynamics in prey composition and temperature conditions. The recruitment and residency of bluefish in the estuary (1998–1999) and ocean beaches (1998), for example, was coincidental with the presence of the Atlantic silverside Menidia menidia and bay anchovy Anchoa mitchilli, the principal prey species for bluefish occupying these respective habitat-types. Bluefish abundance in the estuary (2000) and ocean beaches (1999–2000) was also correlated with water temperature, with the greatest catches of juveniles coinciding with their optimal growth temperature (24 °C). Bluefish growth, estimated as the slope of age–length relationships and daily specific growth rates, equaled 1.27–2.63 mm fork length (FL) d⁻¹ and 3.8–8.7% body length increase d⁻¹, respectively. The growth of sagittal otoliths was also used as a proxy for changes in bluefish size during and shortly before their time of capture. Accordingly, otolith growth rates of summer-spawned bluefish were greater at ocean beaches relative to the estuary and were explained by the more suitable temperature conditions found at ocean beaches during the mid- to late summer. Notwithstanding the fast growth of oceanic summer-spawned bluefish, individuals spawned in the spring were still larger in absolute body size at the end of the summer growing season (240 and 50–200 mm FL for spring- and summer-spawned bluefish, respectively). The size discrepancy between spring- and summer-spawned bluefish at the onset of autumn migrations and during overwintering periods may account for the differential recruitment success of the respective cohorts.     

饲料添加多维对欧洲鳗鲡(Anguilla anguilla)生长性能、消化酶活性和免疫的影响

采用单因子试验设计方法,研究了饲料中添加多维对欧洲鳗鲡(Anguilla anguilla)生长性能、消化酶活性和免疫的影响。试验用鳗(5.93±0.02)g,分6组(每组3重复,每个重复30尾鱼),分别投喂添加不同浓度(0%,0.25%,0.50%,0.75%,1.00%,1.25%)的多维饲料42d。结果表明,随着多维添加水平的上升,各组鳗鲡增重率和特定生长率呈先升高后降低趋势,且差异显著(P0.05),当多维浓度为0.75%时增重率达最高峰值。鳗鲡的肝体指数和肥满度受多维影响较小,在各组间均无显著差异(P0.05)。饲料中添加0.75%浓度的多维显著降低了鱼体水分含量,使鱼体粗蛋白和灰分含量得到显著提高(P0.05),但各试验组鳗鲡粗脂肪含量并无显著差异(P0.05)。相比对照组,各添加组鳗鲡肠道胰蛋白酶、淀粉酶、脂肪酶活性及血清超氧化物歧化酶、溶菌酶和碱性磷酸酶活性均得到显著提高(P0.05),且0.75%组鳗鲡血清总蛋白浓度达到最大(P0.05)。综合本研究结果可知多维在饲料中的适宜添加水平为0.75%。     

刀鲚、凤鲚和湖鲚矢耳石的形态学比较研究

选取凤鲚(Coilia mystus)、湖鲚(Coilia nasus taihuensis)以及采自崇明、黄海和瓯江口3个水域的刀鲚(Coilia hasps)的矢耳石为对象,描述了这些鱼类耳石的形态特征,并进一步使用基于12组形态数据的框架测量进行了种间和种内比较和判别分析.结果表明耳石形态的差异种间较种内更为明显...     

海南岛附近海域肉芝软珊瑚的鉴定

肉芝软珊瑚属(Sarcophyton)是珊瑚礁生态系统中常见的软珊瑚,具有极强的药用价值.我国肉芝软珊瑚种类资源十分丰富,但目前国内对此种珊瑚的分类研究处于停滞状态,肉芝软珊瑚外部形态差异较小,难以直接进行种类鉴定.本研究采集海南省三亚市西瑁岛、万宁市甘蔗岛和大洲岛珊瑚礁区的26个肉芝软珊瑚样品,根据骨针形态学初步鉴定,并分析线粒体msh1基因和COI基因序列进行种类鉴定.结果显示:所有采集的26个样品,鉴定为5个种,分别为:Sarcophyton cherbonnieri、Sarcophyton crassum、Sarcophyton trocheliophorum、Sarcophyton glaucum和Sarcophyton ehrenbergi.其中S. cherbonnieri和S. crassum为我国新纪录种.本研究为海南岛附近海域肉芝软珊瑚的鉴定提供参考,并为我国软珊瑚种类鉴定和生物多样性的研究提供基础数据.     

龙须菜(Gracilariopsis)/江蓠(Gracilaria)的生长和种质特性分析

龙须菜(Gracilariopsis lemaneiformis)是具有重要经济价值和生态效益的大型红藻, 主要用作琼胶提取原料和鲍的饵料。本研究利用生理生化、液相色谱-质谱联用和氨基酸分析等方法, 比较了龙须菜(Gp. lemaneiformis) (wt、981、Gl-1、Gl-s、Gl-g)、异枝龙须菜(Gp. heterocla, Gh)和细基江蓠繁枝变种(Gracilaria tenuistipitata var. liu, Gt)的生长及藻胆蛋白、琼胶、红藻糖苷和氨基酸等的差异, 以期为龙须菜/江蓠栽培中的种质区分及选择提供参考。结果表明Gt在23 ℃和30 ℃条件下生长均最快, 其相对生长速率分别为野生型龙须菜(wt)的2.19倍和2.49倍。龙须菜Gl-s中藻胆蛋白浓度最高, 为wt的1.91倍。除了Gt之外, 其余6种藻中琼胶产率较高(16.22%~18.91%)。Gt中红藻糖苷和海藻糖积累最多, 分别为wt的3.50倍和1.81倍。Gl-g、Gl-s、Gt和Gh多糖丰富, 在36.89%~40.23%; 龙须菜981、Gl-1、wt和Gl-s总氨基酸浓度较高, 在152.35~161.32 mg·g^–1干质量之间, 并且981、Gl-1、Gl-s氨基酸评分较优。综合以上结果, 龙须菜981、Gl-1和Gl-s的藻胆蛋白、琼胶和氨基酸等均显著优于其他藻, 并且生长较快, 可用于琼胶、藻胆蛋白及多糖的提取或鲍的饵料; 而细基江蓠繁枝变种生长快、红藻糖苷和海藻糖丰富, 可大规模栽培用作鲍的饵料。该研究为丰富及开发利用中国大型海藻种质资源提供了重要的资料。     

盐度对花鳗鲡(Anguilla marmorata)和太平洋双色鳗鲡(A. bicolor pacifica)幼鳗鳃丝 及肾脏Na+/K+-ATP酶活力的影响

采用酶学分析的方法,以淡水组为对照,进行了盐度变化对花鳗鲡(Anguilla marmorata)[体质量(10.70±0.92)g]和太平洋双色鳗鲡(A. bicolor pacifica)幼鳗[体质量(12.11±0.79)g]鳃丝及肾脏Na+/K+-ATP酶活力影响的研究。结果表明,经不同的盐度(5、10、18)处理,各处理组的花鳗鲡和太平洋双色鳗鲡幼鳗鳃丝Na+/K+-ATP酶活力均呈现先降低后升高最后再降低并趋于稳定的趋势。前者24h时达到最低值,48h时盐度10处理组上升至与对照组无显著差异(P>0.05),而盐度18处理组显著高于对照组(P<0.05)。后者6h时达到最低值,12h时盐度5和18处理组达到最高值,24h时盐度10处理组达到最高值。盐度变化对花鳗鲡和太平洋双色鳗鲡幼鳗肾脏Na+/K+-ATP酶活力影响不明显。太平洋双色鳗鲡幼鳗鳃丝Na+/K+-ATP酶酶活提升速度及提升幅度均强于花鳗鲡幼鳗,故认为在盐度范围0—18时,太平洋双色鳗鲡幼鳗比花鳗鲡幼鳗对盐度变化具有更强的适应能力。     

太平洋双色鳗鲡(Anguilla bicolor pacifica)致病性类志贺邻单胞菌的鉴定和中药敏感性研究

从发病死亡的养殖太平洋双色鳗鲡(Anguilla bicolor pacifica)肝脏中分离到一株优势菌AMSH1,为研究其种属、毒力及中药敏感性,结合BIOLOG微生物鉴定系统和16S r DNA序列分析对该菌株进行鉴定,并通过人工回归感染试验确定其致病性。此外,还研究了10味中药单用、10种双联用和4种三联用对该菌株的体外抑菌作用。结果显示该菌株为类志贺邻单胞菌(Plesiomonas shigelloides)。经人工腹腔注射感染健康太平洋双色鳗鲡,证实该菌具有致病性,半致死量LD50为3.2×105CFU/g。药敏试验结果表明,10味中药均有较好的抑菌效果,其中五倍子(Galla chinensis)、丁香(Eugenia caryophllata)和生地榆(Sanguisorba officinalis)的效果最好,7种双联用复方和1种三联用复方均具有协同抑菌作用。本次引起太平洋双色鳗鲡发病的病原为类志贺邻单胞菌,该菌对太平洋双色鳗鲡的致病性及其中药药敏的研究属首次报道。建议选用含五倍子的中药双联用复方或三联用复方进行防治,皆绿色环保。     

巴基斯坦国日本金线鱼（Nemipterus japonicus）种群动力学的研究

Japanese threadfin bream Nemipterus japonicus(Bloch, 1791) is among the most abundant and commercially important species in Pakistan. From the coast of Pakistan, four demersal trawl surveys in October–November 2009 and May–June, August, October and November in 2010 were carried out. The purpose of this study is to estimate the population dynamics and status of the stock of the N. japonicus from Pakistani waters based on the research trawl surveys from the research area. The data consist of n=784 length-weight pairs and n=7 530 length frequency with the maximum length and weight of 29 cm and 358 g respectively. The length frequency data were analyzed using ELEFAN method in FiSAT computer package. The parameters of length and weight relationship were b=2.778, a=0.032 and R2=0.973. The estimated von Bertalanffy growth function parameters were L∞=30.45 cm, K=0.270 year-1. Based on length-converted catch curve analysis the total mortality(Z) during this study was estimated at 0.960 year-1. The natural mortality coefficient(M) was 0.74 year-1 using Pauly's equation(the annual average sea surface temperature was 27°C), therefore, the fishing mortality coefficients(F) were 0.22 year-1. The yield per recruit analysis indicated that when tc was 2, Fmax was estimated at 1.2 and F0.1 at 1.1. When tc was 1, Fmax was estimated at 0.95 and F0.1 at 0.8. Because current age at first capture is about 1 year and Fcurrent was 0.22, Fcurrent is smaller than F0.1 and Fmax, which indicated that the fishery is about in a safe condition. When using Gulland(1971) biological reference point, Fopt was equals to M(0.74). The current fishing mortality rate of 0.22 was smaller than the target biological reference point.     

日本鳗鲡补充群体的日龄、全长、出生时间及耳石生长的变化

在中国大陆沿岸6个取样床搜集了日本鳗鲡白仔鳗,191尾用于长度测定,112尾鉴定了日龄及测定了耳石半径,近中心核的标记轮直径、耳石生长轮的宽度.补充群体长度平均值从(53.4±2.2)mm到(58.1±1.7)mm,长度与补充群体的地理分布之间无内在联系.补充群体日龄从(138.2±6.7)d至(155.2±9.0)d,长江口和温州地区补充群的日龄最低,以此为中心,向高纬度和低纬度走向分布,补充群的日龄都逐渐增大,补充群出生时间从1993年8月15日至1994年1月10日.低纬度的群体补充的时间早,其出生时间也早.纬度越高,补充的时间越迟,出生时间也晚.补充群耳石半径平均值从(142.5±8.4)μm至(155.8±14.5)μm,具有显着性差异(p<0.01).标记轮直径从(10.36±0.85)μm至(10.81±1.26)μm,其差异不具显著性(p>1.01).耳石生长轮平均宽度为(0.996-1.097) μm,其差异具有显著性(p<0.01),生长轮平均宽度以温州和长江口群体最大,以此为中心,向高纬度和低纬度分布的补充群的平均值分别逐渐减小.补充群耳石从中心至边缘生长轮宽度变化的共同点是在第90轮处平均宽度值最高.     

中国东南海域拟菱形藻属的新记录种和产毒种

拟菱形藻属（Pseudo-nitzschia）物种是一类备受关注的海洋有毒有害硅藻,我国相关研究仍然不足,已报道的物种多样性偏低,产毒物种也较少。为了解我国海域拟菱形藻属的物种多样性,阐述产毒物种信息,在我国东南海域建立了8个拟菱形藻单克隆培养株系,结合形态学和分子系统学数据,鉴定到2个物种:环孔拟菱形藻（Ps.circumpora C.H.Lim,C.P.Leaw&P.T.Lim）和多列拟菱形藻[Ps.multiseries（Hasle） Hasle],其中环孔拟菱形藻是我国的新记录种。利用高效液相色谱-质谱联用法检测了藻株的多莫酸（domoic acid,DA）特征,在5个多列拟菱形藻株系中检测到DA,单细胞产毒水平为1.01~96.11 fg。此外,利用卤虫（Artemia salina L.）和强额拟哲水蚤（Paracalanus crassirostris Dahl）对多列拟菱形藻的产毒株系（MC4177）进行诱导,发现能够提高其单细胞产毒水平,提高幅度在26~55倍。     

基于线粒体COI基因序列的4个海域口虾蛄群体的遗传多样性研究

为比较中国不同海域口虾蛄(Oratosqilla oratoria)群体的遗传特性, 作者对采自皮口、绥中、青岛和广州4 个海域的口虾蛄群体的线粒体COI基因序列进行扩增和测序, 比较并分析了其遗传多样性和系统进化关系。获得585bp 的口虾蛄线粒体COI基因序列, 发现变异位点54 个, 占总位点数的9.2%。COI基因序列A+T 含量(64.8%)显著高于G+T 含量(35.2%), 符合节肢动物线粒体DNA 碱基组成的特点。转换与颠换的平均比值是7.84, 碱基替换未达到饱和。100 个样本的线粒体COI基因序列共定义35 个单倍型, 单倍型多样性(Hd)为0.9162, 平均核苷酸多样性(π)为0.0203。4 个群体均具有高的单倍型多样性和低的核苷酸多样性的特点, 说明4 个海域口虾蛄的遗传多样性均处于中等水平, 但广州海域的遗传多样性水平最高。AMOVA 分析表明, 来自于群体间的遗传差异(84.53%)明显高于来自群体内的差异(15.47%)。遗传分化系数(F_st)表明皮口、绥中、青岛3 个海域间几乎没有发生分化(F_st<0), 而广州海域口虾蛄遗传分化较大。从GenBank 上下载了19 条粤东海域口虾蛄的同源序列与本实验获得序列共同构建NJ 系统发育树, 结果显示皮口、绥中、青岛聚为一支, 广州和粤东(深圳和汕尾)聚为另一支。单倍型系统发育树和单倍型进化网络关系图均显示出广州海域口虾蛄群体较大的遗传分化。