用包埋脱水法冰冻保存小新月菱形藻

采用包埋脱水法研究了小新月菱形藻(Nitzschia closterium minutissima)的超低温保存,探讨了影响存活率的内、外因素:藻细胞年龄、脱水速率、胶球含水量和化冻后的恢复方法等。结果表明,将处于静止初期的藻细胞包埋在含有0.5 mol/L蔗糖的3%褐藻酸钙胶球中,以含水量每小时减少0.9%的平均速率脱水至胶球含水量为40%,化冻后在冻存管中室温下暗恢复12 h的条件下冰冻存活率较高,可达到18.3%左右。与常规的两步法冰冻保存相比,包埋脱水法可大大简化操作程序,是保存小新月菱形藻的一种有潜力的方法。     

温度对2种饵料金藻保存效果的影响

研究了在室温25℃±2℃,4,-5,-22℃下绿色巴夫藻(Pavlova viridis)和球等鞭金藻(Isochrysis galbana)的保存效果,通过测定其存活率以及主要脂肪酸的变化情况筛选出了2种金藻的适宜保存温度。结果表明,绿色巴夫藻短期内(30~45 d)在-5℃下保存效果最好,较长期(45 d以上)保存则应在-22℃,而球等鞭金藻则在-22℃下保存效果最好。     

用包埋脱水法冰冻保存牟氏角刺藻

采用包埋脱水法研究了牟氏角刺藻(Chaetaceros muelleri)的超低温保存，探讨了影响存活率的内、外因素：藻细胞年龄、蔗糖浓度、脱水速率、胶球含水量等。结果 表明，将处于静止初期的藻细胞包埋在含有0．5mol／L蔗糖的3％褐藻酸钙胶球中，以平均速率0．9％含水量／h脱水至胶球含水量为40％的条件下冰冻存活率较高，可达到30％左右。与常规的两步法冰冻保存相比。包埋脱水法可大大简化操作程序，是保存牟氏角刺藻的一种有潜力的方法。     

包埋脱水法超低温保存萱藻(Scytosiphon lomentaria)丝状体的研究

以本实验室保存的萱藻(Scytosiphon lomentaria)丝状体为材料,用包埋脱水法超低温保存萱藻丝状体,研究了蔗糖预培养浓度和时间、胶球含水量、化冻温度以及胶球恢复时间对萱藻丝状体存活率和冻存后生长发育能力的影响。结果显示:(1)萱藻丝状体胶球在0.4mol/L的蔗糖溶液中预培养6h能获得最高的存活率;(2)萱藻丝状体胶球的最佳含水量较低,为15%左右,当含水量高于27%时,冻存后的存活率为0;(3)40°C为最适的化冻温度;(4)化冻后,将萱藻丝状体胶球在黑暗条件下恢复18h能显著提高存活率;(5)在最佳条件下,萱藻丝状体经冻存后的存活率最高可达54.79%,恢复后的丝状体与冻存前丝状体并无区别,且经诱导后能够产生正常的孢子囊,放散的孢子可以发育成叶状体。     

球等鞭金藻3011浓缩细胞低温保存技术的研究

为了探讨球等鞭金藻3011（Isochrysis galbana Parke）浓缩细胞的保存条件,通过研究其在不同温度条件下的长期保存效果来确定最佳的保存温度范围。实验结果表明,在0～10℃的低温下长期（2个月以上）保存后,球等鞭金藻3011获得较高的存活率（70％以上）,并且在恢复生长时保持较高的比生长率,因此0～10℃是球等鞭金藻3011浓缩细胞长期保存的最佳温度范围。     

包埋脱水法冷冻保存裙带菜配子体克隆的研究

将裙带菜配子体克隆包埋在含3%褐藻酸钙的胶球中,研究了预培养中的蔗糖浓度和培养时间,脱水过程中的脱水速率和胶球含水量以及化冻速度对配子体克隆的存活率和发育能力的影响,结果表明含裙带菜配子体克隆的胶球在0.3 mol/dm3的蔗糖培养基中培养9 h可以获得较高的存活率;胶球最适平均脱水速率为含水量的5%每小时,最适含水量为25%;较快的化冻速度可以显著提高配子体克隆的存活率,在优化条件下存活率可高达66%;经过冷冻保存的裙带菜配子体克隆在适合发育的条件下能够正常形成孢子体.     

一株海洋金藻的18S rRNA 基因序列分析及分类

Isochrysis sp.strain HG是一株分离自福建长乐自然海区的金藻,是一种良好的西施舌育苗饵料藻。显微形态观察表明,Isochrysis sp.strain HG的形态特性与等鞭金藻属的球等鞭金藻(I.galbana)和湛江等鞭金藻(I.zhanjiangensis)比较接近。进一步克隆Isochrysis sp.strain HG的核糖体小亚基(small subunit,SSU)18S rRNA基因,获得了长度为1780bp的基因序列。同源性分析表明,该序列与在NCBI数据库中登录的等鞭金藻属的18S rRNA基因序列同源性最高,说明结果与形态鉴定的结果相一致。基于18S rRNA基因序列的系统发育分析表明,Isochrysis sp.strain HG与等鞭金藻属的I.zhangjiangensis、Isochrysis sp.strain CCAP927/14、I.galbana和Isochrysis sp.strain Santou聚在一个分支类群上,其中I.zhangjiangensis与Isochrysis sp.strain CCAP927/14聚为一个独立的子类群,和I.galbana、Isochrysis sp.strain Santou以及目标菌株Isochrysis sp.strain HG形成4个并列独立的分支。根据形态及分子系统分析结果,Isochrysis sp.strain HG可能是不同于I.zhangjiangensis、I.galbana的等鞭金藻种类。     

温度和微藻饵料对强额孔雀水蚤摄食的影响

论文以强额孔雀水蚤(Parvocalanus carssirostris)为研究对象,研究了不同培养温度(18℃、22℃、26℃、30℃)、不同种类微藻(球等鞭金藻(Isochrysis galbana)、牟氏角毛藻(Chaetoceros muelleri)、青岛大扁藻(Platymonas helgolandica...     

超声波对湛江等鞭金藻生长和脂肪酸组成的影响

设计了一个超声频率、超声功率和处理时间三因素四水平的正交实验,研究了超声波对湛江等鞭金藻的生物学效应.实验结果表明,超声频率对湛江等鞭金藻的生物学效应显著,而超声功率和处理时间的影响相对较小.对于提高湛江等鞭金藻生长速率常数,最佳超声条件为20kHz,6W,10s3次;对于提高其脂肪酸不饱和度和主要不饱和脂肪酸百分比,最佳超声条件为20kHz,4～6W,30s.在最佳超声条件下,湛江等鞭金藻生长速率常数最高可达0.630d,是对照组的2.02倍,其脂肪酸不饱和度最高可达79.6%,比对照组提高7.8%,其中主要不饱和脂肪酸的百分含量均得到不同程度的提高.     

两种海洋单胞藻浓缩与保存效果的研究

研究了小球藻和球等鞭金藻分别用明矾和石灰水浓缩以及两种藻的浓缩液在常温(20 ±1℃)、低温(0—4℃)﹑冷冻(-30±1℃)三种温度条件下保存的结果,并对浓缩保存前后藻液的饵料效果进行了对比试验。 结果表明:(1)小球藻的浓缩以80±5ppm的明矾液及4%的石灰水效果最好;(2)球等鞭金藻的浓缩以100±10ppm的明矾液及6%的石灰水效果最好; (3)保存方法以加入保护剂甘油并置于-30℃ 冰箱中效果最好,小球藻和球等鞭金藻的存活率分别为95%和93%;(4)低温保存前后藻的脂肪酸分析结果表明高度不饱和脂肪酸(HUFA)的含量变化不明显;(5)用浓缩保存藻投喂中国对虾和轮虫的效果与普通藻无显著差异。     

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.     

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.     

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.     

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.