环境因子对硬壳蛤Mercenaria mercenaria稚贝成活率和生长率的影响

采用室内模拟的方法，对盐度、饵料、底质和温度对硬壳蛤稚贝成活率和生长率的影响进行了研究。结果表明，硬壳蛤稚贝存活和生长的适宜盐度范围为18－42，最适盐度范围为24－36。从稚贝成活率看，几种单胞藻的饲育效果顺序为：等鞭金藻＞美国扁藻＞混合藻＞亚心形扁藻＞大溪等鞭金藻＞三角褐指藻＞小球藻＞中肋骨条藻；但从稚贝日生长率看，几种单胞藻的饲育效果顺序为：混合藻＞等鞭金藻＝美国扁藻＝亚心形扁藻＞大溪等鞭金藻＞三角褐指藻＞小球藻＞中肌骨条藻。硬壳蛤对底质环境有一定的选择性，稚贝在纯砂底质中的成活率和日生长率均高于在纯海泥底质中，但砂的粒径大小对稚贝成活率和日生长率没有明显影响。硬壳蛤稚贝具有很强的耐高温能力，在室内控温条件下，能够耐受30－35℃的高温，但在室外自然温度条件下，当温度为21．8－35．4℃（平均为30．1℃）时，稚贝成活率却很低，无砂情况下只有32％，有砂情况下为零。     

长牡蛎两种engrailed同源基因的鉴定及其与贝壳发育相关性的研究

engrailed基因属于同源异形基因家族成员,在许多动物的分节、附肢发育、神经系统发育和贝壳形成过程中发挥作用。本研究克隆了长牡蛎两个engrailed同源基因,命名为cgi-eng1和cgi-eng2。序列分析表明,两个基因均具备典型engrailed基因保守的5个EH结构域。利用整装原位杂交技术检测了cgi-eng1和cgi-eng2在贝壳形成的关键时期早期D形幼虫时期的表达情况。结果显示,cgi-eng1和cgi-eng2 m RNA高表达于贝壳外缘,可能与早期贝壳形成过程有关。此外,两个基因的在贝壳外缘的表达模式亦有区别,提示两个基因的功能可能存在一定程度的分化。本研究首次系统鉴定了长牡蛎engrailed基因的成员,并发现它们可能均参与幼虫贝壳形成,研究结果有助于加深对贝类早期发育及贝壳形成的理解。     

温度对硬壳蛤Mercenaria mercenaria(Linnaeus，1758)呼吸排泄的影响

于 2 0 0 2年 1 1月— 2 0 0 3年 4月 ,采用Winkle滴定法和次溴酸盐氧化法对硬壳蛤呼吸排泄进行了研究。结果表明 ,在 5— 35℃温度范围内 ,温度和体重对硬壳蛤 (壳长 0 5— 6cm ,软体部干重 0 0 0 34— 0 5 9g)的耗氧率和排氨率均有显著影响 (P <0 0 5 ) ,其单位体重耗氧率在2 0— 30℃之间达到最大值 ,且随着硬壳蛤个体的增大而下降 ;不同温度下硬壳蛤个体耗氧率OR[mgO2 /(ind·h) ]与软体部干重W(gDFW)之间的关系可用方程OR =aWb 表示 ,其中a值在0 0 4— 0 70之间 ,b值在 0 45— 0 65之间。硬壳蛤的排氨率 温度曲线呈单峰型 ,其单位体重排氨率在 2 5℃附近达到最大值 ,且随着硬壳蛤个体的增大而下降。不同温度下硬壳蛤个体排氨率NR[μgNH4 N/(ind·h) ]与软体部干重W(gDFW)之间的关系可用方程NR =aWb 表示 ,其中a值在 1— 2 7之间 ,b值在 0 40— 0 75之间。硬壳蛤呼吸排泄的氧、氮比在 5℃时最大 ,随后有下降趋势 ,其比值在 1 0— 80之间。     

副溶血弧菌攻毒过程中文蛤肝胰腺弧菌载量变化的分析

文蛤(Meretrixpetechialis)是我国一种重要的海水养殖贝类,其养殖过程中病害导致的规模性死亡时有发生,已有的工作证实致病性弧菌是导致文蛤大规模死亡最为常见的病原。在本研究中,我们通过浸泡感染实验模拟了文蛤在自然条件下的弧菌胁迫环境,明确了攻毒水体中弧菌生长变化规律;攻毒过程中宿主载菌量分析显示,文蛤肝胰腺组织弧菌载量在攻毒后呈第1天急剧上升,第3天迅速下降的变化趋势,而且不同个体间载菌量差异较大;通过不同弧菌含量攻毒实验,发现在攻毒早期文蛤体内弧菌载量与水体环境的弧菌含量之间呈显著的正相关(Spearman’sρ=0.899,P=0.000),而在感染中后期不同攻毒强度组之间无显著差异,且呈现较低水平,为0～205CFU/mg,预示着宿主免疫系统和肝胰腺微生物群落可将致病菌的数量控制在一定的范围。上述研究为开展文蛤感染发病过程中弧菌载量和免疫抗性评价相关研究提供了参考。     

不同花纹文蛤(Meretrix meretrix)的ITS2分析

采用分子生物学方法,进行了江苏和广西两个地区不同花纹文蛤群体的核糖体DNA转录间隔区(ITS2)的序列分析研究。结果表明,广西的文蛤群体和江苏的群体相比较,仅发现江苏群体在333—336bp处发生了缺失,江苏群体内不同花纹文蛤个体间的ITS2也发生了变化。聚类分析结果证实了序列分析结果,广西文蛤群体和江苏文蛤群体间发生了遗传变异,江苏文蛤群体内部也发生了遗传分化。     

海湾扇贝样品不同保存条件下DNA的提取及RAPD扩增比较

以海湾扇贝为材料，研究了鲜活样品以及采用冷冻保存、70％乙醇固定和10％福尔马林固定等方法保存的闭壳肌样品用于DNA提取的不同效果，并分析了不同保存条件下所提得DNA用于RAPD扩增的结果，研究表明，鲜活样品以及采肜冷冻保存和70％乙醇固定的样品可以得到很好的DNA，其质量和得率没有显著差异，利用上述DNA模板进行RAPD分析，可以获得一致性很好的扩增结果。10％福尔马林休固定样品则没有得到可用的DNA模板。采用70％乙醇固定保存海洋生物组织，是用于DNA提取及相关的分子生物学研究的快速、简易和有效的方法。     

紫扇贝与海湾扇贝种间杂交的研究

为推动中国扇贝养殖业的进一步发展,作者于2008年从秘鲁引进了紫扇贝(Argopecten purpuratus),尝试将紫扇贝与海湾扇贝(Argopecten purpuratus)进行杂交,以期培育出具有优势性状的杂交子代.利用引进的紫扇贝与海湾扇贝进行种间杂交,培育出紫扇贝(雌)×海湾扇贝(雄)杂交扇贝.实验证明紫扇贝的卵子与海湾扇贝的精子可正常受精,受精卵发育正常.杂交扇贝的生长速度在幼虫阶段和成体阶段均快于海湾扇贝.至收获时,杂交贝比海湾扇贝平均壳高、壳长和壳宽分别大27.3%,25.2%和19.3%,体质量增加107.0%,杂种优势明显.     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.     

The phylogeny of native and exotic scallops cultured in China based on 16S rDNA sequences

Scallops of the Family Pectinidae are a valuable resource in marine industry of the world. Understanding the phylogeny of the family is important for the development of the industry. In this study, partial 16S mitochondrial rDNA genes were obtained from 8 scallop species that are commonly cultured indigenous and transplanted species in China. Phylogenetic relationships of Pectinidae were analyzed based on the 8 sequences and other 5 published ones in GenBank, representing 9 genera of the family. The molecular phylogeny trees were constructed using 3 methods with software PHYLIP. The results showe that total 13 species of scallops clustered in 4 clades. Pecten maximus joins P. jacobaeus then Amusium pleuronectes in cluster, indicating close relationship of genus Amusium with Pecten in evolution. P. yessoensis is close to Chlamysfarreri and C. islandica. No enough material was available to single out genus Patinopecten as an independent monophyletic subfamily. The position ofAdamussium colbecki indicates that it is far from genus Pecten but near to genus Chlamys in evolution.     

Survival, growth and immune activity of scallop Chlamys farreri cultured at different depths in Haizhou Bay (Yellow Sea, China) during hot season

Survival, growth and immune response of the scallop, Chlamys farreri, cultured in lantern nets at five different depths (2, 5, 10, 15, and 20 m below the sea surface) were studied in Haizhou Bay during the hot season (summer and autumn) of 2007. Survival and growth rates were quantified bimonthly. Immune activities in hemolymph (superoxide dismutase (SOD) and acid phosphatase (ACP)) were measured to evaluate the health of scallops at the end of the study. Environmental parameters at the five depths were also monitored during the experiment. Mortalities mainly occurred during summer. Survival of scallops suspended at 15 m (78.0%) and 20 m (86.7%) was significantly higher than at 2 m (62.9%), 5 m (60.8%) or 10 m (66.8%) at the end of the study. Mean shell height grew significantly faster at 10 m (205.0 μm/d) and 20 m (236.9 μm/d) than at 2, 5 or 15 m in summer (July 9 to September 1); however, shell growth rate at 20 m was significantly lower than at the other four depths in autumn (September 2 to November 6). In contrast to summer, scallops at 5 m grew faster (262.9 μm/d) during autumn. The growth of soft tissue at different depths showed a similar trend to the shell. Growth rates of shell height and soft tissue were faster in autumn than in summer, with the exception of shell height at 20 m. SOD activity of scallops increased with depth, and ACP activity was significantly higher at 15 and 20 m than at other depths, which suggests that scallops were healthier near the bottom. Factors explaining the depth-related mortality and growth of scallops are also discussed. We conclude that the mass mortality of scallop, C. farreri, during summer can be prevented by moving the culture area to deeper water and yield can be maximized by suspending the scallops in deep water during summer and then transferring them to shallow water in autumn.