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。     

分离草虾杆状病毒包涵体的一种新方法

本文报道了一种纯化草虾杆状病毒（MBV）包涵体的方法。取出受MBV感染的虾肝胰组织，在TESP溶液中（50mmol/dm^3Tris-HC1 pH8.0,50mmol/dm^3EDTA,0.1mol/dm^3NaC1,0.5mmol/dm^3PMSF）破碎，差速离心，再经30%～70%蔗糖浓度梯度高速离心，可得到含包涵体的区带。电镜下观察到较纯的包涵体，经蛋白酶K降解后，PCR扩增，经琼脂糖凝胶电泳可见清晰的DNA区带。     

一种提取眼点拟微球藻(Nannochloropisis oculata)DNA的简便方法

用氯化锂法从眼点拟微球藻(Nannochloropisis oculata)中提取DNA并对其提取条件进行了优化。新鲜藻细胞在提取液(0．6mol／L LiCl，0．8％Sakosyl，20mmol／L EDTA(pH8．0)，0．4％PVP，10mmol／L Tris-HCl(pH8．0)，5％β—巯基乙醇)中经过55℃温育30min，每mg鲜藻可提取20—40mg DNA，提取的DNA分子量在23kb左右，可用于PCR、限制性酶切等。     

三唑磷对四种水生生物的毒性及安全评价研究

本文研究了三唑磷对小球藻、卤虫、南美白对虾、泥蚶四种水生生物的急性毒性作用，结果显示，三唑磷对小球藻的48h的EC50为24μg/dm^3,72h的EC50为15μg/km^3；对卤虫的24hLC50为1．64mg／dm^3，48hLC50为0．8mg／dm^3；对南美白对虾仔虾的48hLG0为3．2μg／dm^3，96hLC50为1．1μg/dm^3。；对泥蚶的48hLC50为21．0mg／dm^3，96hLC50为10．2mg／dm^3。三唑磷对小球藻、卤虫、南美白对虾为高毒农药，泥蚶对三唑磷有很强的耐受力。     

不同保存条件下中华哲水蚤基因组DNA提取的比较

以采自厦门港的中华哲水蚤(Calanus sinicus)为对象，研究多种保存条件对中华哲水蚤基因组DNA提取的影响。结果表明，除5％福尔马林保存的样品没有提取到DNA之外，其余样品均能成功提取出基因组DNA；以该DNA为模板通过相应引物成功扩增出线粒体DNACOI基因片段。其中无水乙醇保存样品提取可靠的基因组DNA方法的建立，为野外样品保存工作提供了一个简便，经济的途径。     

黑鱾血液的生理生化指标研究

本文采用F-820半自动血细胞分析仪和全自动生化分析仪,对黑纪的血液常规指标及血清中的17项生化指标进行了分析测定,并用Wright’s染色法制作血涂片对白细胞进行分类计数．结果表明：（1）黑纪的血红细胞数量为（2．49±0．47）×10^12个／dm^3;白细胞数量为（1．08±0．21）×10^11个／dm^3;在白细胞中以嗜中性粒细胞数量最多,占52．42％±4．67％,其次为淋巴细胞,占25．72％±2．88％;单核细胞体积最大,占总数的3．27％±0．62％．（2）黑纪红细胞脆性大,抗性差,在0．65％～0．70％的NaCl溶液中开始溶血,在0．50％～0．55％的NaCl溶液中完全溶血．（3）黑纪的血红蛋白（HGB）为109．11±15．72g／dm^3,高于一般鱼类,与其较强的耐低氧生理能力相适应．（4）血清中血糖浓度为4．72±1．35mmol／dm^3,明显低于凶猛肉食性鱼类,与其岛礁性生态习性和杂食性相吻合．（5）雌雄鱼的乳酸脱氢酶分别为332．67±188．48U／dm^3和905．66±257．85U／dm^3,雌鱼显著高于雄鱼,可能与繁殖活动相关．     

用于单-藻细胞PCR扩增反应的样品保存方法比较

用室内培养的塔玛亚历山大藻（Alezandrium tamarense ATHK藻株）作为实验对象,分别选择了5％福尔马林固定后常温保存、95％乙醇固定后常温保存、-20℃冷冻保存、鲁哥氏液（Lugol’s solution）固定后常温保存等方法,在5,15,30,60d后对保存的样品进行单细胞PCR反应,并与新鲜样品进行比较。实验结果表明,用95％乙醇保存、-20℃冷冻保存、鲁哥氏液保存的样品在60d后仍可扩增出目标条带,与新鲜样品没有差别,但是样品中的藻细胞形态有一些改变;而以5％福尔马林固定保存的样品只能在5d后扩增出目标条带,但藻细胞形态比较完好。因此,对用于单细胞PCR实验的微藻样品,短期内（不多于5d）可以采用福尔马林保存,而需要长期保存的样品,应当采用乙醇或鲁哥氏液固定,或者采用-20℃冷冻保存的方法。     

福建外来船舶压舱水中浮游植物种类组成与丰度及其影响因素的初步研究

2006～2008年间对进入福建沿海4个主要港口的12艘国际航船所携带的压舱水进行浮游植物种类组成和丰度分布的调查.此调查共检测出浮游植物7个门86属239种（含变种和变型）,77μm孔径网滤样和20μm孔径网滤样的平均藻类丰度分别为1.2×10^2cells/dm^3（变动在0～9.1×10^2cells/dm^3间）和3.4×10^3cells/dm^3（变动在0～3.0×10^4cells/dm^3间）.同时对其中6艘船舶的压舱水样品用f/2培养基培养,共有13种硅藻和1种甲藻被成功培养.文中还分析了压舱水中浮游植物丰度与盐度、水龄的关系,并结合历史资料讨论了压舱水里各类浮游植物的分布.     

单条固定线虫基因组DNA提取及18S rRNA基因PCR扩增

根据线虫18S核糖体RM基因PCB扩增效果比较了丙酮、乙醇、乙醇 0．05mol／L FDTA(pH8．0)和5％海水福尔马林4种固定剂,碱裂解和蛋白酶K处理2种单条线虫基因组DNA提取方法的优劣。用乙醇固定的样品最适合制备RR模板DNA,而5％海水福尔马林固定的样品能最完整地保持样品形态。蛋白酶K处理获得的DNA较碱裂解获得的更适合PCR扩增。结果有助于分子生物学方法在海洋线虫分类、多样性和生态学研究中的应用。     

高密度轮虫对点带石斑鱼早期仔鱼培育影响的研究

本试验在水体为1000dm^3、仔鱼密度为15ind．／dm^3条件下，研究不同轮虫密度(10、25、40、70、100ind．／cm^3)对点带石斑鱼早期仔鱼摄食和生长的影响．至仔鱼12日龄的培育试验结果表明，在点带石斑鱼早期仔鱼的培育过程中，投放轮虫对水质有影响，特别是在2～6日龄间，DO值和pH值都有一降低过程，但随后即回升并相对稳定；轮虫密度越高则pH值越低，pH值最低出现在4～6日龄，4日龄时10ind．／cm^3组的pH值比100ind．／cm^3组高0．32．在10～100ind．／cm^3的轮虫密度范围内，较低的轮虫密度对仔鱼的生长有利，12日龄10ind．／cm。组仔鱼的全长比100ind．／cm^3组高31．83％，但100ind．／cm^3组仔鱼的成活率比10ind．／cm^3组的高出近一倍；不同轮虫密度下仔鱼摄食轮虫数量的差异并不明显；但9日龄以后，过高的轮虫密度100ind．／cm^3下仔鱼生长速度显著降低．     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

A Review of Some Environmental Issues Affecting Marine Mining

This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested.     

Increased Particle Fluxes at the INDEX Site Attributable to Simulated Benthic Disturbance

Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement.     

Determination of In Situ Sediment Shear Strength from Advanced Piston Corer Pullout Forces

The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments.     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.