厚壳贻贝(Mytilus coruscus)组织及其海水环境中真菌群落结构差异分析

海洋真菌广泛参与近海生态系统的物质循环和能量流动, 同时与海洋动物之间存在复杂的相互作用。贝类是我国主要的海水养殖生物, 为深入了解海洋真菌与贝类养殖的潜在关系, 选择厚壳贻贝养殖区海水及8种组织真菌为研究对象, 利用荧光定量PCR以及ITS rDNA高通量测序解析养殖厚壳贻贝各组织及所处海水环境的真菌群落丰度和结构特征。结果显示厚壳贻贝养殖区内和边缘海域的真菌丰度显著高于养殖区外围海域; 从贻贝养殖区和组织中共获得1 409个OTUs, 其中粪壳菌纲(Sordariomycetes) 在海水真菌群落为优势纲; 而在贻贝组织中, 锤舌菌纲(Leotiomycetes, 足20.13%、肾脏14.72%)、座囊菌纲(Dothideomycetes, 鳃2.89%、后闭壳肌1.92%、血淋巴1.36%)、散囊菌纲(Eurotiomycetes, 性腺3.59%、足1.57%)和伞菌纲(Agaricomycetes, 性腺3.09%、消化腺2.71%、鳃2.50%)占据优势地位。多样性分析显示厚壳贻贝和海水间真菌群落存在显著性差异; Bray-Curtis相似距离分析显示贻贝真菌群落与养殖区内海水更为相似, 而与边缘和外围海水差距较大。厚壳贻贝不同组织间、不同区域海水间的Beta多样性差异主要来自物种替换; 贻贝与海水间真菌Beta多样性的差异主要来自丰富度差异。综上所述, 厚壳贻贝体内真菌具有组织差异性, 并且养殖活动改变了养殖区海水的真菌群落。研究结果将为贝类真菌资源、贝类-真菌相互作用及生态影响提供基础。     

紫贻贝养殖产业的现状、问题与对策

为全面系统地了解中国紫贻贝(Mytilus galloprovincialis)养殖产业情况,作者从养殖产量、养殖面积、进出口贸易、养殖生产等方面概述了紫贻贝产业现状,对中国紫贻贝养殖业的种质资源、苗种生产方式、养殖生产流程、养殖方式等进行了综述,并针对紫贻贝养殖中存在的问题,从种质资源、养殖模式、设施设备、食品安全控...     

THE GENETIC DIVERSITY OF CULTURED MYTIL US CORUSCUS POPULATIONS BASED ON SEQUENCE ANALYSIS OF MITOCHONDRIAL CO Ⅲ GENE

基于线粒体DNA的COⅢ基因序列对我国沿海重要经济贝类厚壳贻贝3个养殖群体（温州、宁德、福州）的遗传多样性和遗传结构进行了分析。由PCR扩增获得23个体的CON基因787bp的部分序列,其多态性遗传参数统计显示,23个个体共检出21个单倍型,总群体单倍型多样性指数（Hd）为0．978,核苷酸多样性指数（Pi）为0．01045,平均核苷酸差异数（的为8．534,三个群体均显示出较丰富的遗传多样性。遗传分化系数（Fst）表明,福州群体与宁德群体和温州群体间有一定程度的遗传分化,而宁德群体和温州群体间无遗传分化。     

不同保存方法对厚壳贻贝样品蛋白提取效果的比较

为了解厚壳贻贝(Mytilus coruscus)稚贝样品在冷冻保存过程中的蛋白变化情况,作者采用直接冷冻法、添加海水冷冻法、蛋白裂解液冷冻法等分别对保存1周至6个月的厚壳贻贝样品蛋白提取的不同效果进行了比较。SDS-PAGE胶结果显示,3种冷冻保存方法均出现4个有规律条带(a、b、c、d),其相对分子量依次约为55、46、38和26kDa。SDS-PAGE蛋白条带灰度值的检测结果表明,1个月内蛋白裂解液法保存的样品最佳;直接冷冻法和添加海水冷冻法的研究发现保存过程中的蛋白均出现不同程度的降解现象,因而可能不适宜用于该种稚贝样品的保存。     

低盐度形成的微生物膜对厚壳贻贝稚贝附着的影响

为研究环境因子在厚壳贻贝(Mytilus coruscus)稚贝附着过程中的调控作用, 作者探讨了低盐度对微生物膜生物构成、群落结构的影响及所形成微生物膜对厚壳贻贝稚贝附着的影响。在实验室条件下, 研究微生物膜的日龄与干质量、细菌密度和硅藻密度、叶绿素a含量的关系及其对厚壳贻贝稚贝附着的影响。通过DGGE指纹图谱技术对微生物膜中的细菌群落结构多样性进行了分析。研究发现,盐度13和23时形成的微生物膜能有效促进厚壳贻贝稚贝的附着, 且盐度23、28d时稚贝附着率最高, 达到72%。相关性分析表明, 微生物膜的诱导活性与盐度、干质量、细菌密度、硅藻密度、日龄呈显著正相关性, 与叶绿素a无相关性。微生物膜的干质量、附着细菌密度及底栖硅藻密度明显随着日龄的增加而增加, 叶绿素a含量与微生物膜日龄无显著相关性。细菌群落在厚壳贻贝稚贝附着过程中发挥重要调控作用。     

Using tolerance intervals to assess recovery of mussel beds impacted by the Exxon Valdez oil spill

Polycyclic aromatic hydrocarbons (PAH) have been measured in mussel tissues in early spring and summer since 1993 throughout Prince William Sound (PWS) and the Gulf of Alaska (GOA). Season-specific thresholds were established at reference sites to identify ‘above background’ total PAH levels. Thresholds were estimated using one-sided 99% tolerance limits. Thresholds were similar across reference sites but differed by an order of magnitude across seasons. Trends in total PAH since 1998 were assessed for sites impacted by the 1989 Exxon Valdez oil spill or the Alyeska Marine Terminal. Summer samples exhibited no trends; early spring samples declined. In early spring, all sites were judged ‘recovered’ by 2004; in summer, one site in western Prince William Sound and two in the western GOA exceeded thresholds by 11 ng/g dry weight or less. Robust estimation methods prevented bias from observations affected by unknown releases or laboratory errors.     

紫贻贝(Mytilus edulis)与厚壳贻贝(M.coruscus)杂交的细胞学观察及子一代的早期生长比较

为了研究紫贻贝与厚壳贻贝种间杂交的核相变化,并评估其杂交优势,本实验以两种贻贝亲本为材料建立了4个交配组合,分别为:紫贻贝自交组EE(Mytilus edulis♀×M.edulis♂)、厚壳贻贝自交组CC(Mytilus coruscus♀×M.coruscus♂)、正交组EC(M.edulis♀×M.coruscus♂)和反交组CE(M.coruscus♀×M.edulis♂)。通过Hoechst 33258染色、荧光显微观察的方法,对4个交配组从受精、减数分裂到卵裂的核相变化过程进行了观察,并对孵化后幼虫到稚贝期的生长情况进行了记录。结果表明:正反交组的受精细胞学过程与自交组较为相似,但第二极体排出时间延长。对各实验组子一代早期的生长和存活研究表明:浮游阶段,杂交组幼虫生长优势不明显;附着变态后,正交组EC在壳长及壳高等生长指标的杂交优势表现明显,且正交组(EC)大于反交组(CE)。自交组的受精率、孵化率、成活率均高于杂交组。     

Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction

Since it is often difficult to estimate possible adverse effects due to contamination in selected ecosystems, multi-species biomonitoring may provide more information, taking into account different routes of exposure, ecological roles and metabolic capabilities of animals. In this context, we exposed for 7 days the mussel Mytilus galloprovincialis and the crab Carcinus aestuarii to 4-nonylphenol (NP), a well-known xenoestrogen. In mussels (0–0.2 mg NP l⁻¹), we measured NP bioaccumulation in soft tissues and vitellogenin (Vg)-like protein levels in digestive glands from both males and females by the alkali-labile phosphate assay (ALP). As no reference data were available for crab exposure, the NP 96-h LC₅₀ value was previously determined. Then, in sublethally exposed (0–1.0 mg NP l⁻¹) male crabs, NP bioaccumulation and Vg levels were measured in hemolymph, gonads and digestive gland. Bioaccumulation of NP increased from 43 to 371 μg g⁻¹ d.w. in mussels, and from 3.6 to 37 μg g⁻¹ d.w. in crabs, depending on the NP concentration in water. Dose-dependent Vg-like protein induction was observed in both species, appearing to be related to NP bioaccumulation, although a partial decrease was recorded at the highest concentration tested. A similar trend was observed in both digestive gland and gonad of exposed crabs; Vg increased to a lesser extent, although significantly, in hemolymph. Results demonstrated that NP induces Vg synthesis both in male and female mussels, as well as in male crabs. On the basis of the responsiveness of both species investigated, a multi-species approach is indicated in biomonitoring programmes.     

Long term monitoring for oil in the Exxon Valdez spill region

In the aftermath of the 1989 Exxon Valdez oil spill, a Long Term Environmental Monitoring Program (LTEMP) has been regularly sampling mussels (and some sediments) for polycyclic aromatic and saturated hydrocarbons (PAH and SHC) at sites in Port Valdez, Prince William Sound, and the nearby Gulf of Alaska region. After 1999, a decreasing trend appears in total PAH (TPAH) in tissues at all sites with current values below 100 ng/g dry weight (many below 50 ng/g). Currently, most samples reflect a predominantly dissolved-phase signal. This new low in TPAH likely represents ambient background levels. Synchrony in TPAH time-series and similarities in the hydrocarbon signatures portray regional-scale dynamics. The five inner Prince William Sound sites show similar composition and fluctuations that are different from the three Gulf of Alaska sites. The two Port Valdez sites represent a unique third region primarily influenced by the treated ballast water discharge from the Alyeska Marine Terminal. Prince William Sound has reverted to a stable environment of extremely low level contamination in which local perturbations are easily detected.     

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed.Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2–3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.