PCB uptake and accumulation by oysters (Crassostrea virginica) exposed via a contaminated algal diet

Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 μg polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills+mantle and muscle of oysters fed algal paste containing 1.0 μg PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills+mantle tissues compared to the same tissues in the undosed control.     

单体牡蛎诱导变态的研究

为了得到壳型规则、大小均一的单体牡蛎,作者以葡萄牙牡蛎(Crassostrea angulata)和长牡蛎(Crassostrea gigas)眼点幼虫为材料,从肾上腺素(EPI)浓度梯度、处理时间梯度和眼点幼虫密度梯度3个因素诱导产生不固着变态的单体牡蛎。结果表明,葡萄牙牡蛎用EPI处理24 h的最适浓度为5×10–4 mol/L,不固着变态率为72.8%,在该最适浓度下,最佳处理时间为12 h,不固着变态率为82.7%。长牡蛎用EPI处理6 h的最适浓度为5×10–5 mol/L,不固着变态率为53.2%,在该最适浓度下,最佳处理时间为8h,不固着变态率为56.8%,眼点幼虫密度在80个/m L以下EPI处理效果没有显著性差异。对长牡蛎幼虫进行后续生长测定,结果显示EPI处理组幼虫的壳长、壳高和存活率要高于对照组,表明EPI可能促进牡蛎幼虫变态长出次生壳并提高其生存能力。     

Cellular responses of oysters, Crassostrea virginica, to metal-contaminated sediments

Elevation of metal concentrations in coastal environments associated with anthropogenic enrichment pose a significant threat to estuarine organisms. The purpose of this study was to evaluate the relationships between cellular responses that may be potentially valuable as indicators of chronic stress and metal-contaminated sediments. For these studies, hatchery-reared juvenile oysters were deployed in situ at 15 sites for approximately 1 month around Charleston Harbor, SC. The effects on lysosomal destabilization and glutathione concentrations were determined; and the relationships between the cellular responses and sediment metal concentrations were described. Both single metal and multiple metal parameters (based on total metal concentrations, aluminum normalizations, and summed sediment quality guidelines) were considered. Generally, significant correlations were observed for individual metal analytes and multiple metal parameters. Since many of the individual metal analytes covary, the responses may reflect overall contaminant loading rather than responses to individual metals. Methods for estimating overall contaminant loading based on multiple analytes provide a more realistic estimate of potential adverse effects.     

Differential recruitment of introduced Pacific oysters and native mussels at the North Sea coast: coexistence possible?

Pacific oysters (Crassostrea gigas Thunberg 1793) have been introduced into the Wadden Sea (North Sea), where they settle on native mussel beds (Mytilus edulis L.), which represent the only extensive insular hard substrata in this soft-sediment environment. As abundances of C. gigas rose, some mussel beds became increasingly overgrown with oysters, whereas others did not. Field experiments revealed that recruitment of C. gigas was higher in the lower intertidal than in the upper subtidal zone, that it was higher on conspecifics than on mussels, and that it was not affected by barnacle epigrowth except when settling on mussels. Mussel recruitment is known from inter- and subtidal zones. It occurred equally on oyster and mussel shells but showed a clear preference for barnacle epigrowth over clean shells. Assuming that settlement and recruitment are key processes for species abundances on the North Sea coast, it is predicted that the positive feedback in oyster settlement will lead to rapid reef formation of this invader at the expense of mussel beds. Mussels, however, may escape competitive exclusion by settling between or on the larger oysters especially when barnacles are abundant. Experimental patches with mussels were more often covered by fucoid algae (Fucus vesiculosus forma mytili Nienburg) than patches with oysters, and oyster recruitment was poor underneath such algal canopies. Thus, fucoids may provide the native mussels with a refuge from the invading oysters and the two bivalves may coexist, provided food is not limiting.     

长牡蛎(Crassostrea gigas)微卫星多重PCR 体系构建及其在家系鉴定中的应用

筛选多态性高、特异性好、片段大小适中的10 个长牡蛎微卫星位点进行优化组合, 根据扩 增片段大小及同一荧光不重叠原则构建了两组五重PCR 体系。运用CERVUS 3.0 软件对0527 组27 个长牡蛎全同胞家系的643 个子代和0612 组27 个全同胞家系382 个子代分别进行亲权鉴定。结果 发现, 用两组微卫星五重PCR 鉴别时, 在0527 组和0612 组的鉴定成功率均为100%; 只用第一组微 卫星五重PCR, 可以将0527 组96%的子代和0612 组96%的子代鉴定到亲本; 只用第二组微卫星五 重PCR, 可以将0527 组97%的子代和0612 组95%的子代鉴定到亲本。本研究中筛选出的两组微卫 星五重PCR 体系在两组家系中的鉴定效率均较高, 可以快速有效地将子代个体鉴定至所属父母本, 在长牡蛎家系鉴定中具有很好的应用价值。     

多肽菌素在长牡蛎人工育苗生产中的应用

多肽菌素作为一类具有抗菌活性的生物短肽,是一种可以替代抗生素的生物肽。为了推广多肽菌素在水产养殖中的应用,本研究以长牡蛎(Crassostreagigas)人工育苗为基础,以添加2×10_–6 mol/L青霉素钠为对照组,以添加5个不同浓度(2×10_–6、4×10_–6、6×10_–6、8×10_–6、10×10_–6 mol/L)多肽菌素为实验组,研究多肽菌素在长牡蛎人工育苗生产中的应用。结果显示:(1)在长牡蛎种贝培育方面,培育前8天各组差异不显著(P> 0.05),培育后期添加6×10_–6 mol/L多肽菌素组存活率均最高,对照组最低,且培育至32天时该实验组均显著高于其他组(P<0.05)。(2)在长牡蛎幼虫培育方面,幼虫壳高日增长量在添加6×10_–6 mol/L多肽菌素组中最大,为11.57μm/d,而对照组仅为9.61μm/d;幼虫存活率方面对照组始终最低,而添加6×10_–6 ...     

长牡蛎早期胚胎发育中脂肪酸和氨基酸组成变化的研究

采用生物化学方法研究了长牡（Crassostrea gigas）早期胚胎发育过程中脂肪酸、氨基酸组成变化情况,分别获取受精后0、6、12、24h的胚胎或幼虫进行检测和分析,结果如下：在受精卵中共检测到27种脂肪酸,豆蔻酸（14：0）、软脂酸（16：0）、硬脂酸（18：0）,EPA（20：5ω3）和DHA（22：6ω3）...     

长牡蛎(Crassostrea gigas)EST串联重复序列的组成和分布

长牡蛎(Crassostrea gigas)串联重复序列分析研究较少,为了研究其在基因组转录本的基本结构特征并为长牡蛎中遗传多样性研究中提供有益的信息,利用NCBI公共数据库中的57 139条长牡蛎ESTs序列对串联重复序列结构类型、分布、丰度等进行系统比较分析.分析结果表明:1)长牡蛎EST中共有小卫星串联重复序列8...     

牡蛎体内及其养殖水体中细菌耐药性研究

为进一步了解牡蛎体内外细菌的耐药性，对分离自阳江的香港牡蛎（Crassostrea hongkongensis）养殖水体及其内脏团的637个异养细菌进行了10种抗生素的药敏试验。采用纸片扩散法（Kirby-Bauer，简称K-B），参照NCCLS抗生素敏感试验操作标准，研究细菌的耐药概况。结果显示：健康牡蛎体内细菌对恩诺沙星和庆大霉素耐药率较低，发病牡蛎体内细菌对庆大霉素耐药率最低，次之为复合磺胺和恩诺沙星；水体细菌对氯霉素、恩诺沙星、环丙沙星耐药率较低，水体及体内细菌对呋喃唑酮耐药率最高，而对卡那霉素及青霉素耐药率则波动较大。结果还显示，不同来源的受试菌株多重耐药状况严重，来源于健康和病牡蛎的菌株在7月份的多重耐药率达到峰值，分别为66.7%和64.3%，养殖水体细菌多重耐药率峰值出现在4月份，最高达76.7%。总体来看，源于牡蛎养殖环境水体及其牡蛎体内的异养菌多重耐药比例较高，值得引起注意。而受试菌株对不同药物的抗性存在较大差异，且异养细菌对呋喃唑酮的耐药率明显高于对其它受测抗生素的耐药率。从时间动态上看，水体菌耐药率峰值出现于4月份，而牡蛎体内菌群耐药率峰值出现于7月份。     

不同长牡蛎群体对海水酸化的生理响应差异分析

海水酸化暴露可对海洋生物产生多层面的影响。本研究以潮间带野生与潮下带养殖长牡蛎(不同生境背景)的不同组织(鳃、外套膜及消化腺)为研究对象,分析在室内调控p CO2模拟海水酸化暴露条件下,其基础代谢活动、能量代谢以及氧化应激相关指标的变化情况。结果显示:海水酸化暴露后,两种长牡蛎(Crassostrea gigas)的基础代谢过程均受到了一定抑制作用且受影响程度差异明显。潮间带野生与潮下带养殖长牡蛎的关键生理过程(能量代谢及氧化应激)对海水酸化暴露存在不同的响应变化,表明两种长牡蛎应对海水酸化的调节机制可能存在差异。依据PLS-DA分析结果显示,在所有生理指标中,对样本的差异贡献较高(VIP值>1)的指标为:SDH、AST、ATPase、ATP含量、糖原含量、CAT、GST及SOD,表明海水酸化暴露后,在两种长牡蛎的3种组织中上述指标的响应变化程度更大。综合评价分析多个生理指标的整体变化揭示:在海水酸化暴露条件下,潮间带野生长牡蛎比潮下带养殖长牡蛎对海水酸化的生理响应更为剧烈;相比于鳃及消化腺组织,长牡蛎外套膜组织可能受影响更大。     

长牡蛎外套膜色遗传规律初步研究

比较了长牡蛎(Crassostrea gigas)不同外套膜色家系的受精率与孵化率,不同生长阶段的生长与存活及外套膜色在子代个体中的分布规律。结果表明,不同家系的卵径、受精率和孵化率无显著差异(P0.05);壳高在60和150日龄稚贝阶段略有差异(P0.05),在350和450日龄成体阶段差异不显著(P0.05);不同生长阶段的存活率差异不显著(P0.05)。外套膜纯黑BB家系的子代外套膜基本全部为4级着色;外套膜纯黄YY1家系子代的外套膜基本全部为1级着色,YY2家系子代的外套膜黄色与黑色比例约为3∶1,符合孟德尔分离定律。由此推测长牡蛎外套膜色可能由一个主效基因控制,且黄色对黑色具有显性上位效应。研究结果可以为长牡蛎外套膜色的人工选育提供理论依据。     

长牡蛎中国群体与日本群体杂交子一代的生长和存活比较

以中国长牡蛎(C)和日本长牡蛎(J)为材料,进行了4个组合的杂交和自交实验,分析了各实验组受精率、孵化率,幼虫和稚贝的生长、存活以及杂交子代的杂种优势。结果表明,自交组的受精率介于杂交组之间,但是经单因素方差分析表明没有显著性差异; JJ组的孵化率最高,其次是JC和CC组, CJ组的孵化率最低,相同卵子来源的组合在孵化率上无显著差异。JJ组在幼虫期和稚贝期、养成期的壳高、壳长平均值均大于其他三组; JC组在幼虫期和稚贝期、养成期的壳高、壳长仅次于JJ组,其壳高、壳长和存活率杂种优势率均较为明显; CJ组的杂种优势率在幼虫期和稚贝养成期的各个生长性状上都表现为负值。杂交子一代中存在正交和反交结果不同的现象可能与母性效应、遗传因素和环境等因素有关。     

盐度胁迫对香港牡蛎部分生化指标的影响

为探究盐度胁迫下香港牡蛎(Crassostrea hongkongensis)生化指标的变化规律,了解盐度适应过程中牡蛎的代谢机制,本研究以盐度0、8、16、32、40为胁迫盐度,以正常海水(盐度24)为对照,开展香港牡蛎对盐度胁迫的响应研究。结果显示,各实验盐度组糖原含量在盐度胁迫0～8 h内下降,且盐度胁迫幅度越大糖原含量降幅越大,胁迫8 h后则无明显的变化规律。腺苷酸激活的蛋白激酶(AMPK)、组蛋白/蛋白去乙酰化酶(SIRT1)和Na~+/K~+-ATP酶活力的变化规律相似:在0～8 h内,各实验组酶活力均急剧下降(SIRT1上升);8～48 h内,酶活力上升;48～120 h内,酶活力逐渐趋于平稳状态,总体表现为盐度越高,酶活力越强,并与胁迫前有明显差异,且盐度胁迫幅度越高,差异越明显。总抗氧化能力(T-AOC)总体表现出高盐胁迫下随时间的增加而升高,低盐胁迫下随时间的增加而降低,且盐度胁迫幅度越大,T-AOC活力的变化幅度越大。实验结果初步表明,香港牡蛎糖原含量与渗透压调节存在一定的关系,AMPK、SIRT1、Na~+/K~+-ATP酶活力及T-AOC均与渗透压调节密切相关,且在高盐胁迫下随时间的增加而升高,低盐胁迫下随时间的增加而降低。     

Spatial variability in growth and reproduction of the Pacific oyster Crassostrea gigas (Thunberg, 1793) along the west European coast

The Pacific oyster Crassostrea gigas was introduced in Europe for commercial purposes in the mid 1960s. It was initially thought that low winter temperatures would restrain this species' reproduction and settlement; however, its present distribution in areas where no introduction has taken place suggests that natural invasion and expansion has occurred. Along the European coast, wild populations of Pacific oysters are already found from northern Germany to southern Portugal. Whether C. gigas will continue to further expand through northern waters will depend on its physiological performance. In this study, the performance of wild oyster populations has been studied in terms of growth and reproduction at three stations: La Rochelle (France; 46°N), Yerseke (Oosterschelde estuary, The Netherlands, 51°N), and Texel (Wadden Sea estuary, The Netherlands, 53°N). The French population had the lowest somatic-shell mass ratio and an increase in maximum shell length, somatic and gonadal mass was observed from France to the Netherlands. In addition, mean oocyte diameter decreased significantly from south to north. The combination of increasing gonadal mass and decreasing oocyte volume suggests an increasing reproductive output in terms of egg numbers from France to The Netherlands. Differences in temperature between locations will at least be partly responsible for the observed patterns; however, other environmental factors (such as food availability, predation pressure, sediment type and/or seston concentration) cannot be excluded. Since smaller eggs (oocytes) are thought to have a longer development time, the environmental conditions along the Dutch coast may result in increased larval dispersal and possibly in further population expansion.     

长牡蛎F3 代快速生长选育群体生长特性的研究

采用模型拟合方法研究了长牡蛎(Crassostrea gigas)F3代快速生长选育群体不同时期各生长性状的发育规律。结果表明,长牡蛎幼虫期壳高(SH)、壳长(SL)对日龄(t)的回归遵循Logistic模型,生长方程分别为SH=455.612/(1+9.500e-0.142t),R2=0.999;SL=462.476/(1+8.026e-0.108t),R2=0.996。幼虫期壳高与壳长成直线相关,回归方程为SL=0.76SH+18.82,R2=0.994。长牡蛎养成期各生长性状呈现明显的季节变化,壳高(SH)、壳长(SL)、壳宽(SW)和总质量(TW)对月龄(X)的多项式回归方程分别为SH=-0.0297X4+1.0365X3-12.0220X2+57.6500X-68.9260,R2=0.985;SL=-0.0173X4+0.5893X3-6.5702X2+30.2420X-34.4150,R2=0.986;SW=-0.0068X4+0.2620X3-3.2806X2+16.9170X-22.1410,R2=0.956;TW=-0.0219X4+0.8234X3-10.1680X2+50.7040X-85.4110,R2=0.972。壳高、壳长、壳宽与总质量均呈幂函数相关,回归方程分别为SH=23.645TW0.3213,R2=0.998;SL=12.337TW0.3776,R2=0.995;SW=6.611TW0.3589,R2=0.981。     

利用四倍体与二倍体杂交规模化培育全三倍体太平洋牡蛎苗种

利用引进美国的四倍体太平洋牡蛎(Crassostrea gigas)雄贝与我国的二倍体雌贝杂交，规模化培育全三倍体牡蛎苗种。杂交组的平均受精率为48．27％，对照组为80．53％，杂交组较低的受精率可能与四倍体雄贝的精子活力及精子数量有关。杂交组的孵化率、幼虫成活率和变态率分别为81．10％，42．3％和1．82％，对照组分别为79．96％，37．0％和1.80％。杂交组共培育出稚贝1200万粒。不同时期取样的流式细胞仪分析结果表明，杂交组的三倍体率为100％。     

Rapid invasion of Crassostrea gigas into the German Wadden Sea dominated by larval supply

Invasions of non-indigenous species into coastal habitats have been a frequent phenomenon in the last decades, sometimes with significant impact on the receiving ecosystem. However, the understanding of the entire process and especially the relative importance of larval supply and local recruitment remains unclear. In this study, we simulate the invasion of a benthic invertebrate into a previously uncolonised habitat over several years and validate the results with field data. Therefore, we present field data from a monitoring programme revealing the rapid invasion of the oyster Crassostrea gigas into the East Frisian Wadden Sea, North Sea, between 2003 and 2005. The applied model combines a simple, spatially-explicit population dynamics model for the adult stage with a particle tracking model for the larval stage of the life cycle. Simulation results are able to reproduce the large-scale pattern of the field data and indicate a domination of larval supply on the population dynamics in the early stage of the invasion. Though monitoring and simulations suggest a single larval source outside the study area in the west, the population dynamics in the eastern part is only explainable with an additional source within the study area attributed to an unintentional input of juveniles by mussel fishery. High sensitivities to uncertain parameters result in distinct deviations between monitoring and simulations at particular sites. Especially the impact of site-specific variations of the post-settlement mortality underlines the variability of local recruitment conditions and indicates the need for spatially resolved information for exact predictions.     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...     

温度与盐度对福建牡蛎与长牡蛎、熊本牡蛎、香港巨牡蛎种间杂交子代孵化及其早期生长成活的影响

本研究以野生福建牡蛎(A)为亲本一方, 分别与野生长牡蛎(G)、熊本牡蛎(S)、香港巨牡蛎(H)进行杂交, 除A♀×S♂、A♀×H♂两组杂交不育外, 共得到AG(A♀×G♂)、GA(G♀×A♂)、SA(S♀×A♂)、HA(H♀×A♂)、AA(A♀×A♂)5个不同组合, 并研究不同盐度与温度对其幼虫孵化率、早期生长及成活率的影响。结果表明, 盐度25时AG、GA、SA、HA、AA五组孵化率分别达到最大值78.33%、75.00%、63.33%、55.00%、71.67%; 温度低于25℃时, GA孵化率高于其他组, 温度高于25℃时AG孵化率最大, 其中25℃时达到最大孵化率83.33%; 不同盐度条件下D型幼虫经过7 d培育, HA在盐度20时壳高达到最大值97.8 µm, 成活率46.67%, 在盐度为35时第5天全部死亡。AG、GA、SA、AA四组在盐度为25时壳高有最大值, 分别为124.7 µm、121.2 µm、93.7 µm, 111.8 µm; 不同温度条件下D型幼虫经过7 d培育, 在温度为25℃时各组壳高和成活率均有最大值, 大小排列依次为AG>GA>AA>HA>SA, 各组成活率随温度升高呈现出先增大后减小的趋势, AG在25℃时成活率在所有组中最高, 为88.89%; 孵化后在适宜温度盐度条件下连续培养16 d, 不同组之间壳高表现为: AG > GA> AA > HA > SA, AG壳高、壳高SGR及成活率始终大于其他组, 而SA、HA两组壳高、壳高SGR及成活率始终小于自交组AA。研究表明, 相较于福建牡蛎与熊本牡蛎、香港巨牡蛎的杂交, 以及福建牡蛎的自交, 福建牡蛎与长牡蛎的杂交子代表现出明显的生长及存活优势, 生长潜力较好, 且AG优于GA; AG组在盐度25、水温25℃条件下生长速率最大, 成活率最高。研究结果可为福建牡蛎育苗行业良种选育提供参考依据。     

长牡蛎nacrein 基因的克隆、结构及进化分析

首次在长牡蛎(Crassostrea gigas)中克隆得到一种新的贝壳基质蛋白nacrein-like protein F3的全长c DNA序列。nacrein-like protein F3基因c DNA全长1499bp,其中编码区长度为1242bp,编码一条含413个氨基酸残基的多肽链。氨基酸序列比对和结构域分析均表明其为合浦珠母贝(Pinctada fucata)nacrein的同源蛋白,含有1个保守的α-碳酸酐酶结构域,但由于重复结构域的插入,α-碳酸酐酶结构域被间隔成2个亚结构域。系统进化分析显示nacrein-like protein F3与贻贝(Mytilus californianus)nacrein-like protein进化关系最近。此外,在软体动物中,双壳纲nacrein-like proteins进化速度相对较快,推测与寒武纪时期剧烈的环境变化有关,如影响贝壳形成的海水化学变化。