栉孔扇贝血细胞的显微和超微结构

通过光镜、扫描电镜和透射电镜观察栉孔扇贝Chlamys farreri血细胞的形态、结构特征,对其分类进行了初步研究。根据细胞的大小、形态和结构特征可将血细胞分成5种类型:Ⅰ型小透明细胞,大小约2.44±0.11μm,约占45%—50%;Ⅱ型大透明细胞,大小约4.83±0.28μm,约占15%—20%;Ⅲ型小颗粒细胞,大小约4.07±0.15μm,约占15%—20%;Ⅳ型中颗粒细胞,大小约7.20±0.26μm,约占20%—25%和Ⅴ型大颗粒细胞,大小约13.87±0.73μm,约占3%—5%。血细胞在血淋巴中的平均密度为(3.03±0.11)×107cell.ml—1,其中颗粒细胞占42.6%,透明细胞占57.4%。透射电镜观察结果表明,颗粒细胞的细胞质颗粒可区分成3种类型:Ⅰ型高电子密度颗粒,Ⅱ型低电子密度颗粒和Ⅲ型中等电子密度颗粒。     

繁殖和高温对栉孔扇贝抗氧化能力的影响

本研究以栉孔扇贝(Chlamys farreri)为对象,通过分析CuZnSOD和MnSOD基因表达水平的变化,探讨繁殖和高温对扇贝抗氧化能力的影响。研究显示:CuZnSOD和MnSOD在栉孔扇贝各组织中广泛表达,2种SOD的表达水平在不同组织间均差异显著(P0.05)。在性腺中CuZnSOD的表达水平显著低于在鳃和肾脏中的表达水平(P0.01),在肝胰腺中MnSOD的表达水平显著高于在鳃、性腺、闭壳肌和肾脏中的表达水平(P0.001)。单因素方差分析显示:繁殖后,CuZnSOD的表达水平在6种组织中均显著降低(P0.05),MnSOD的表达水平仅在鳃和肾脏中显著下降(P0.05);经过1个月的恢复期及再次繁殖后,2种SOD的表达水平与繁殖后相比均未呈现显著差异。高温胁迫实验表明:30℃胁迫12.5h,扇贝死亡率达到62.28%;此时,CuZnSOD的表达水平在鳃、性腺、肾脏及闭壳肌中显著下降,MnSOD的表达水平在外套膜、鳃和肾脏中显著下降。结果显示:繁殖和高温均导致扇贝多个组织中CuZnSOD和MnSOD的表达水平受到不同程度的影响,其中,鳃和肾脏中2种SOD的表达水平在繁殖和高温胁迫后均显著下降,表明繁殖和高温严重影响了这2种组织的抗氧化能力。研究结果将为揭示海洋双壳类动物夏季大规模死亡的原因提供参考。     

雌性虾夷扇贝与雄性栉孔扇贝杂交子代及其亲本的核型研究

采用滴片法制片-吉姆萨染色对雌性虾夷扇贝(Patinopecten yessoensis)与雄性栉孔扇贝(Chla-mysfarreri)杂交子代及其双亲的核型进行了研究,报道了在扇贝杂交过程中产生的特殊的细胞遗传学现象,结果表明,栉孔扇贝染色体核型2n=38=6m+10sm+22st;虾夷扇贝染色体核型2n=38=6m+10sm+16st+6t;两者杂交子代染色体数目2n=38,但其染色体组型不符合预期型,不是简单地由两个亲本各自一套染色体组型组合而成,呈现出复杂的形态多态性,端部着丝点染色体明显增多.     

栉孔扇贝DAPI带型和PI带型研究

选取栉孔扇贝Chlamys farreri担轮幼虫为材料,采用秋水仙素-低渗-空气干燥法制备染色体标本,应用荧光显带技术,分析了DAPI带和PI带在栉孔扇贝染色体上的分布。DAPI带型结果显示,栉孔扇贝所有染色体上都存在DAPI阳性带,主要分布于传统的着丝粒区和端部区域,另外还存在一些中间区DAPI带及可变带,总带数为62。PI带型结果与DAPI带型结果相似,在所有染色体上都存在PI阳性带。2种带型的阳性带所在位置与异染色质分布区域相吻合。     

Application and comparison of two symptom-based eutrophication-assessment methods in Jiaozhou Bay,China

Increasing numbers of approaches to assess eutrophication, such as estuarine trophic status, the Oslo-Paris Commission Common Procedure, and the Water Framework Directive, focus on the symptoms of eutrophication. In China, however, nutrient index methods dominate the assessment of coastal waters. In this study, an integrated method that includes both water quality and ecological response was compared with the Northwest Pacifi c Action Plan (NOWPAP) Common Procedure. Observation data from Jiaozhou Bay, Shandong, China, were used in a comparison of the two methods in a trophic status study. Overall, both clearly revealed a high level of nutrient enrichment in the bay, indicated by high nutrient concentrations. Though the two methods diff ered in their methodological design in the assessment of the ecological eff ects of nutrient enrichment, they have acquired similar results: the integrated method suggested that the status was good, and the NOWPAP Common Procedure suggested that the status was low (indicating that the bay had no serious eutrophication problem). The introduction of fi lter feeders (shellfi sh aquaculture) into the bay on a reasonable scale may have been eff ective in mitigating eutrophic conditions, and perhaps explains the low ecological impacts there. Our results will be useful to ecosystem-based eutrophication management in the bay and in similar areas.     

磁化水在栉孔扇贝人工育苗上的应用研究

本实验以栉孔扇贝为材料，对栉孔扇贝亲贝、幼虫，进行０．０５～０．９Ｔ不同磁场强度的磁化水培育试验．实验结果表明：磁化水对亲贝蓄养有明显促进性腺发育成熟的作用；磁化水组性腺指数比对照组提高３．５×１０－２．孵化率也大大提高，比对照组提高１６×１０－２．对扇贝幼虫生长发育的影响，在０．３～０．７Ｔ的磁化水范围内，幼虫生长较快，成活率最高，其中０．６Ｔ最好。然后又进行０．６Ｔ生产性应用试验，通过３ａ应用，磁化水组比对照组幼虫成活率可提高１８×１０－２，附着变态率比对照组提高２７×１０－２，单位水体出苗量比对照组提高５０×１０－２左右。另外，磁化水也有明显改善育苗木质的作用，从而保证了扇贝育苗稳产高产。     

胶州湾沉积物重金属形态不同浸取方法的比较与污染讯息指示作用初探

海洋沉积物中重金属的活性形态对于指示沉积物污染状况具有重要作用;为探寻简洁且能够有效提取重金属活性形态的浸提方法;实验研究了0.1 mol/L和1 mol/L盐酸的单级提取和欧共体标准局（European Community Bureau of Reference）提出的BCR分级提取法对胶州湾表层沉积物中重金属（Cr、Cu、Ni、Pb、Zn）和Al、Fe、Mn的浸取效果。结果显示;3种提取方法提取的金属含量呈显著正相关关系;两种浓度的稀盐酸对BCR方法中的酸可提取态和可还原态金属均具有较好浸提效果;但对可氧化态金属浸提能力相对较弱。不同提取方法的人为信号指数（ASI）计算结果表明;BCR浸取法提取结果的金属ASI值最大;表明其对沉积物中金属活性形态提取效率较高;1 mol/L HCl提取结果的金属ASI值最低;可能是较高浓度强酸使沉积物较大颗粒内层中“惰性”金属被浸取出来;“增加”了金属活性组分的污染讯息信号;对沉积物中Cu、Pb和Zn而言;0.1 mol/L HCl提取结果的ASI值与BCR方法接近。经过Al归一化后;0.1 mol/L HCl浸提的重金属含量的空间分布与胶州湾的实际污染状况相吻合;表明0.1 mol/L的HCl在一定程度上能够代替复杂的BCR浸取法;此方法可简化重金属活性形态的提取步骤;适合大范围沉积物重金属污染评价。     

海洋生态重要性区域的内涵与识别方法研究——以黄河口为例

海洋生态环境目前正在承受来自人类活动和气候变化的巨大压力;急需必要的管理工具或决策支持系统来应对这些压力所带来的生态环境问题。对海洋生境的分类识别是开展生物多样性保护的基础和前提;海洋生态重要区识别作为一种客观、科学和灵活的生境分类评估和选划方法;已被科学界和管理者广泛接受和认可。本文首先介绍了海洋生态重要区的概念内涵以及识别方法的研究进展;给出了海洋生态重要区的定义;之后以黄河口为例;初步建立了针对该区域的生态重要区识别标准和评价方法;对黄河口及邻近海区进行了生态重要性等级划分。本文是对生态重要区识别方法在我国近海的首次应用尝试;研究结果可为我国基于生态系统的海洋管理提供科学方法和工具;也可为其他区域的研究提供借鉴。     

中国北方沿海6个栉孔扇贝群体的AFLP分析

利用4对AFLP引物首次对我国北方沿海主要的6个栉孔扇贝养殖区的自然采苗群体进行了遗传多样性分析,共扩增得到191条清晰带谱,其中175条具有多态性。对各群体进行遗传多样性分析,结果表明各群体均具有较高的遗传多样性水平,香农式多样性指数在0.347 4～0.391 7之间。AMOVA结果表明遗传变异主要来源于群体内,占总遗传变异的87.22%。个体聚类结果表明,烟台、长岛、大连3个群体的个体聚在一起,可看做1个群体;而荣成群体的个体单独聚在一起,具有独立的遗传结构;胶南和日照的一部分个体聚在一起,其他大部分各群体单独聚在一起;推测是海洋环境和人工养殖影响的共同结果。     

栉孔扇贝Fosmid文库的构建及基因组结构特征分析

研究构建第一个栉孔扇贝的fosmid基因组文库,该文库包含133 851个克隆,插入片断平均长度为40 kb,覆盖栉孔扇贝基因组的4.3倍.栉孔扇贝DNA在fosmid传代中表现得较为稳定,没有发现插入片段的丢失或重排.所有的克隆进行了超级池和二级池的构建,并筛选了2个基因和7个微卫星,结果阳性克隆数介于2到8之间.由此可见,该文库可以很好的用于目的基因或标记的筛选,将有利于物理作图和基因的图位克隆研究.2 016个克隆进行双末端测序,获得3 646 条序列.序列总长2 286 986 bp,大约占栉孔扇贝基因组的1.84‰.共得到2 500条串连重复序列,其中微卫星序列371条,小卫星序列1 816条,卫星序列313条.共得到317条散布重复序列,总长97 412 bp,占测序总长的4.26%.查找到的散布重复序列共有4种类型:DNA 转座子、LTR 反转座子、LINE反转座子和滚环转座子,其中LINE反转座子的数目最多.BLAST比对共有1 383条序列与数据库现有的序列有明显的相似性(E＜e-5),占测序总数的37.93%.BLASTN比对有明显相似性的1 036条序列中,与nr数据库序列相似的有113条,与EST数据库序列相似的有923条.BLASTX比对有明显相似性的序列有347条,占序列总数的9.52%.     

栉孔扇贝(Chlamys farreri) 血细胞类型及抗菌力的研究

用2种方法将栉孔扇贝血细胞分为2大类透明细胞、颗粒细胞.吉姆萨染色观察,透明细胞的细胞质中颗粒很少,细胞核圆形,居中,细胞质较少,核质比大;颗粒细胞比透明细胞大且多,染色深,但核质比小,在细胞质中有颗粒,有些颗粒细胞具双核.密度梯度离心结果,透明细胞主要位于上层,颗粒细胞则大部分位于下层.吞噬力、抗菌活力、溶菌活力及活性氧检测结果表明,颗粒细胞均强于透明细胞.     

胶州湾李村河口沉积物中重金属分布特征及其控制因素

对胶州湾李村河口4个站点沉积物中的酸可挥发性硫化物(AVS)以及同步提取金属元素(SEM,包括Cu,Cd,Ni,Pb和Zn)的含量进行测定,以研究重金属元素在河口海湾沉积物中的分布特征及影响其分布的因素。结果表明,J1站表层沉积物中同步提取金属含量高于其它站点;除J1站外其他站点的同步提取金属的总量随沉积物深度变化范围不大;J1站的SEM/AVS比值始终小于1,其他站点SEM/AVS比值只在沉积物浅表层大于1。此外,AVS和同步提取Fe的含量对沉积物中同步提取金属的分布有着重要影响,其他的因素如有机质的含量等也会对沉积物中同步提取金属的分布产生影响。     

Effects of benzo[a]pyrene on DNA damage and histological alterations in gonad of scallop Chlamys farreri

To investigate the biological impact of polycyclic aromatic hydrocarbons (PAHs) on reproductive system, scallops Chlamys farreri were continuously exposed to benzo[a]pyrene (BaP) (0.5, 3, 10 μg L^?1) during 15 days. DNA damage and histological alterations were examined in female gonad. DNA strand break levels significantly increased after 12 h exposure, and remained high and significantly different from control values until the end of the exposure. In the ovaries of the scallops exposed to 10 μg L^?1 BaP for ten days, histological analysis showed that the cytoplasts of the oocytes in the outer layer of the ovaries became sparse, and the nuclear membranes were obscure. Damaging effects on ovaries and oocytes were more severe after 15 days exposure. Degenerating oocytes increased and the connective tissue of the ovary envelops became loose. Electron microscopic examination revealed ultrastructural aspects of degenerating oocyte and degenerated oocyte after 15 days exposure.     

栉孔扇贝血细胞活性检测方法的筛选与应用

筛选栉孔扇贝(Chalmys farreri)血细胞活性的检测方法,研究苯并[a]芘(B[α]P)对栉孔扇贝血细胞活性的影响.结果表明96孔培养板中栉孔扇贝血细胞接种密度在(1～32)×10~4 cell/孔时,血细胞活性无显著性差异(P>0.05),且细胞存活率在97.42%～98.82%之间,台盼蓝排斥法、四甲基偶氮唑(MTT)、二甲氧唑黄(XTT)比色法、乳酸脱氢酶(LDH)活性测定法均可作为栉孔扇贝血细胞活性的测定方法,而当血细胞接种密度在(12～32)×10~4/孔时,中性红比色法亦可作为血细胞活性的检测方法;同时实验采用Eagle培养液进行血细胞原代培养在0～24 h内血细胞活性较好.采用中性红比色法检测B[α]P对栉孔扇贝血细胞活性具有显著抑制作用,且抑制程度与B[α]P作用浓度、时间呈正相关,其余方法均未检测出B[α]P对血细胞活性的影响.由此可见,B[α]P对栉孔扇贝血细胞活性具有免疫毒性作用,中性红比色法可以作为评价B[α]P对栉孔扇贝血细胞免疫活性的敏感指标.     

Distributions and seasonal variations of dissolved carbohydrates in the Jiaozhou Bay,China

Surface seawater samples were collected in the Jiaozhou Bay, a typical semi-closed basin located at the western part of the Shandong Peninsula, China, during four cruises. Concentrations of monosaccharides (MCHO), polysaccharides (PCHO) and total dissolved carbohydrates (TCHO) were measured with the 2,4,6-tripyridyl-s-triazine spectroscopic method. Concentrations of TCHO varied from 10.8 to 276.1 μM C for all samples and the ratios of TCHO to dissolved organic carbon (DOC) ranged from 1.1 to 67.9% with an average of 10.1%. This result indicated that dissolved carbohydrates were an important constituent of DOC in the surface seawater of the Jiaozhou Bay. In all samples, the concentrations of MCHO ranged from 2.9 to 65.9 μM C, comprising 46.1 ± 16.6% of TCHO on average, while PCHO ranged from 0.3 to 210.2 μM C, comprising 53.9 ± 16.6% of TCHO on average. As a major part of dissolved carbohydrates, the concentrations of PCHO were higher than those of MCHO. MCHO and PCHO accumulated in January and July, with minimum average concentration in April. The seasonal variation in the ratios of TCHO to DOC was related to water temperature, with high values in January and low values in July and October. The concentrations of dissolved carbohydrates displayed a decreasing trend from the coastal to the central areas. Negative correlations between concentrations of TCHO and salinity in July suggested that riverine input around the Jiaozhou Bay had an important effect on the concentrations of dissolved carbohydrates in surface seawater. The pattern of distributions of MCHO and PCHO reported in this study added to the global picture of dissolved carbohydrates distribution.     

胶州湾大型底栖动物的丰度、生物量和生产量研究

为了研究胶州湾大型底栖动物的丰度、生物量和生产量,于2002年3月、6月、8月和12月,在胶州湾北部软底区、大沽河口、黄岛养殖区及养殖区邻域选取4个站位进行采样,对大型底栖动物进行了定量研究。共采到大型底栖动物138种,总平均丰度、平均生物量(湿质量)和年生产量(有机碳)分别为1 719个/m2,27 g/m2,2.2 g/(m2.a),初步估算,胶州湾大型底栖动物的总次级生产量为2.8万t/a。与渤海和南黄海大型底栖动物的丰度和生物量比较,丰度和生物量均低于这两个海域,但是胶州湾大型底栖动物的总次级生产量高于渤海。本研究对于了解胶州湾大型底栖动物现状及湾内养殖对大型底栖动物的影响具有重要的意义。     

栉孔扇贝(Chlamys (Azumapecten) farreri) 消化盲囊的组织学和组织化学的研究

运用石蜡切片法、透射电镜技术及组织化学方法，对栉孔扇贝的消化盲囊进行了研究。结果表明消化盲囊的腺上皮由消化(吸收)细胞和分泌细胞组成，消化(吸收)细胞顶端有微绒毛，细胞内有酸性磷酸酶、碱性磷酸酶、脂酶和酯酶活性，还含有糖原和脂肪。是进行食物的细胞内消化和营养物质吸收的主要场所，并有储存能量的功能；分泌细胞内含粗面内质网、高尔基体和分泌颗粒，具有酸性磷酸酶和碱性磷酸酶活性。并含有丰富的RNA和蛋白质，有分泌消化酶的功能；消化盲囊导管上皮细胞顶端有纤毛和微绒毛，细胞内有酸性磷酸酶、碱性磷酸酶及脂酶活性，也能分泌消化酶。消化(吸收)细胞浅部的细胞质及其释放到盲囊腔中的胞质团中含铁；各部位不含钙。     

A three-dimensional water quality model and its application to Jiaozhou Bay, China

A three-dimensional coupled physical and water quality model was developed and applied to the Jiaozhou Bay to study water quality involving nutrients, biochemical oxygen demand, dissolved oxygen, and phytoplankton that are closely related to eutrophication process. The physical model is a modified ECOM-si version with inclusion of flooding/draining processes over the intertidal zone. The water quality model is based on WASP5 which quantifies processes governing internal nutrients cycling, dissolved oxygen balance and phytoplankton growth. The model was used to simulate the spatial distribution and the temporal variation of water quality in the Jiaozhou Bay for the period of May 2005 to May 2006. In addition, the effect of reduction of riverine nutrients load was simulated and evaluated. The simulated results show that under the influence of nutrients discharged from river, the concentrations of nutrients and phytoplankton were higher in the northwest and northeast of the bay, and decreased from the inner bay to the outer. Affected by strong tidal mixing, the concentrations of all state variables were vertically homogeneous except in the deeper regions where a small gradient was found. Obvious seasonal variation of phytoplankton biomass was found, which exhibited two peaks in March and July, respectively. The variation of riverine waste loads had remarkable impact on nutrients concentration in coastal areas, but slightly altered the distribution in the center of the bay.