伶鼬榧螺(Oliva mustelina)的分子鉴定及其形态变异

根据榧螺螺旋部的差异可将其分为两种不同的形态类型,类型Ⅰ(MorphotypeⅠ)的个体螺旋部陷入体螺层中,其壳顶与前部数螺层愈合;类型Ⅱ(MorphotypeⅡ)的个体螺旋部高出壳顶,缝合线明显,此类型可明确鉴定为伶鼬榧螺(Oliva mustelina)。本研究通过线粒体COⅠ和16SrRNA基因序列的分析对两种形态类型的榧螺进行了分子鉴定。结果表明:两种形态类型的榧螺含有共享的单倍型;不同形态类型间的遗传距离和同一形态类型内的遗传距离重叠;两者的单系性在系统发育分析中没有得到显现,而是共同构成一个单系。因此,两种形态类型的个体均为伶鼬榧螺。类型Ⅰ可能是伶鼬榧螺中一种少见的有别于典型个体的形态变异类型。     

曼氏血吸虫宿主藁杆双脐螺与几种扁蜷螺形态解剖学比较

曼氏血吸虫中间宿主螺藁杆双脐螺和其他3种隶属扁蜷螺科的吸虫及线虫中间宿主螺,其形态极为相似,缺乏分类学资料,比较几种扁蜷螺的形态及生殖系统构造,明确其分类学特征,能为监测及防控部门准确鉴定物种提供依据。电子数显卡尺测量所有样本螺壳参数;解剖4种扁蜷螺生殖系统并绘图;扫描电镜观察齿舌形态。方差分析表明,4种扁蜷螺壳高(H)和直径(D)比值差异显著(F=64.171,P0.01)。4种扁蜷螺齿舌均为中央齿1列,双齿型,但中央齿齿尖形态、侧齿及缘齿列数、齿尖数量及形态等依种类不同均有差异。研究结果表明不同种的齿舌形态是重要鉴别特征之一。生殖系统阴茎复合体构造在属间差异明显,种间较为相似(旋螺属的凸旋螺、小旋螺)。尽管4种扁蜷螺贝壳形态较为相似,但壳高(H)和直径(D)比值、齿舌列数、齿尖以及生殖系统的阴茎复合体构造等特征,对入侵物种藁杆双脐螺和中国几种常见扁蜷螺的准确鉴定有重要价值。     

栉孔扇贝(Chlamys farreri)壳顶幼虫血细胞分布的免疫学观察

研究组前期实验发现,栉孔扇贝血细胞在D形幼虫时期出现.本文应用单克隆抗体的免疫组化、免疫电镜方法进一步定位栉孔扇贝壳顶幼虫的血细胞并观察其分布.结果表明,栉孔扇贝壳顶幼虫已分化出界限明显的组织器官,包括外套膜、面盘、口、食道、胃、消化腺、足等;血细胞主要分布于壳顶幼虫的外套膜、面盘、食道、消化腺、胃等组织器官内及周围,其中面盘、消化腺、胃等处有大量血细胞成簇分布.血细胞形状不规则,直径为3～5μm;细胞核多为圆形、椭圆形,位于细胞的一侧;细胞质内有线粒体、内质网等细胞器和空泡.结果证实,栉孔扇贝在壳顶幼虫时期已出现了大量清晰可辨的血细胞,其分布特点与成贝血细胞相似.     

三种壳口颜色脉红螺（Rapanavenosa）形态学和线粒体16SrRNA与COI基因片段差异比较分析

脉红螺壳口通常有3种颜色：“黑白条纹螺”、“中间螺”和“橙色螺”。本论文对3种壳口颜色的脉红螺分别进行了形态学和mt16SrRNA、COI基因片段序列比较分析。形态学分析结果表明,除厣宽／壳高、厣高／壳高、厣高／体螺层、厣高／壳口长、厣高／壳口宽、厣高／厣宽的形态学特征指数和出肉率不具有显著性差异外,其它形态特征和生物学相关变量具有显著差异,且对湿重的影响效果不同。“黑白条纹螺”壳口长和壳口宽对湿重影响最大,而“中间螺”和“橙色螺”各形态特征对湿重均无显著的直接影响。根据各形态特征均值建立的居群形态聚类图显示,“中间螺”和“橙色螺”形态差异较小,二者与“黑白条纹螺”的形态差异较大。然而基于mt16SrRNA和COI基因片段序列分析结果显示,三种壳口颜色的脉红螺无遗传分化,为同一种。     

疣荔枝螺(Thais clavigera Kuster)人工繁育技术研究

本实验开展了疣荔枝螺（Thais clavigera Kuster）室内全人工繁育技术研究；具体包括亲螺人工促熟蓄养、卵囊采集、孵化、幼虫培育、变态和采苗等技术研究，同时，还开展了后期面盘幼虫对不同附着基的喜好选择性实验，以及不同饵料和光照对幼虫附着的影响研究。研究表明：雌螺分批产卵，单个雌螺平均每次产出受精卵为61 750粒；在水温27~29℃时，面盘幼虫经过20 d左右的生长发育，壳长达到约600 μm，此时开始附着变态，由浮游生活转变为底栖生活；附着后10 d左右变态为稚螺。后期面盘幼虫对附着基的选择性实验表明，固着牡蛎苗的栉孔扇贝壳和附有底栖硅藻的波纹板，是稚螺理想的附着基。不同饵料和光照对幼虫附着影响实验结果表明：在附着变态期间，自然光照有利于幼虫附着变态；同时，连续投喂糠虾肉糜可显著提高幼虫变态率和稚螺成活率。本研究结果为今后疣荔枝螺产业化繁育提供了重要的理论和技术支持。     

海南产芋螺的形态特征

从海南各个市县沿海采集到20多种芋螺（Conus），并制成标本，对这些芋螺标本进行了详细的外部形态特征测量与描述，包括芋螺壳形状、大小、体质量、壳顶特点、表面光滑度、螺纹、色彩及花纹等。鉴定出20种海南产的芋螺种类，为芋螺毒素基因的分离和芋螺毒素药物的开发提供基础。     

皱肋文蛤胚胎、幼虫及稚贝的发育

在水温28.5～30℃、盐度23.6～25.4 条件下, 观察了皱肋文蛤(Meretrix lyrata)胚胎、幼虫和稚贝的发育过程。结果表明, 皱肋文蛤的胚胎经历桑椹期、囊胚期、原肠胚期等阶段, 形成担轮幼虫。根据行为方式和形态特征, 其幼虫阶段可分为D 形幼虫、壳顶初期幼虫、壳顶中期幼虫、壳顶后期幼虫、变态期幼虫、附着期幼虫。稚贝发育可分为无管期稚贝、单管期稚贝和双管期稚贝。皱肋文蛤幼虫在不同阶段的生活习性逐渐改变, 从担轮幼虫至壳顶后期幼虫营浮游生活, 变态期和附着期幼虫分别营匍匐生活和附着生活。壳顶初期幼虫出现足原基, 壳顶中期和壳顶后期幼虫分别形成平衡囊和鳃丝。稚贝阶段开始从附着生活向埋栖生活过渡。稚贝出现明显的次生壳, 无管期稚贝和单管期稚贝的次生壳为红褐色, 而双管期稚贝的次生壳为白色。皱肋文蛤从D 形幼虫发育到附着期幼虫, 后者的壳长和壳高分别增至前者的1.77 倍和2.10 倍; 稚贝阶段从无管期发育到双管期, 后者的壳长和壳高分别增至前者的3.13 倍和3.28 倍, 显示皱肋文蛤幼虫阶段缓慢生长, 稚贝阶段快速生长的特征。     

砗蚝(Hippopus hippopus)的人工繁育

于2016年4—9月开展了砗蚝(Hippopus hippopus)人工繁育技术研究。采用五羟色胺进行催产、促使配子排放;精卵分别收集,进行异体间受精以避免自交;受精卵经过30h孵化,选出600万D形幼虫进行培育。砗蚝的早期生活史与砗磲(Tridacna spp.)相似,经历前期面盘幼虫、中期面盘幼虫、后期面盘幼虫、足面盘幼虫、单水管稚贝、双水管稚贝、外套膜触手稚贝、幼贝等阶段。与砗磲不同的是,砗蚝怀卵量较少,但卵径较大,D型幼虫也较大,幼虫趋光性更强,壳长2.0mm以后外套膜不伸出壳缘外,幼贝贝壳形态也不同于砗磲。同砗磲幼虫一样,砗蚝幼虫需要构建虫黄藻系统之后,才能出现鳃、次生壳等,从而完成变态成为稚贝。砗蚝幼虫变态率较低,仅为1.4%。中间育成期间,丝状藻和锥形螺是稚贝培育的主要敌害,需及时清理才能确保稚幼贝正常生长发育。经过120d的精心饲育,培养出平均壳长6.3mm幼贝500余个。本研究为进一步开展砗蚝人工繁育、中间育成、增殖放流、资源修复及移植保育提供了参考。     

象拔蚌人工育苗技术研究

通过对亲贝促熟、人工催产、幼虫培育和采苗等技术环节的研究,建立了象拔蚌(Panopea gen-erosa)的人工繁育技术。研究表明,在盐度28~30条件下,受精卵在水温11、13、16oC时发育至D形幼虫所需时间分别为83、60和46 h;在水温16~18oC,经20 d发育至壳长约380μm的壳顶幼虫时出现足和水管原基,进入附着变态阶段。投放附着基后培育约70 d,可生长至壳长5 mm的幼贝。壳长日增长量在D形幼虫阶段为6.2μm,在壳顶幼虫阶段为16.8μm,附着变态后生长速度加快,外形具成体形态。不同附着基的采苗率依次为：细砂>卵石>聚乙烯网片>波纹板。     

文蛤(Meretrix meretrix)幼虫发育过程中细胞凋亡和Caspase 功能分析

以文蛤为研究对象,利用TUNEL技术(Terminal-deoxynucleotidyl transferase mediated dUTP Nick End Labeling)检测了文蛤各发育阶段的幼虫中细胞凋亡的部位,利用Caspase原位标记分析了有活性的Caspase在幼虫中的分布情况,用Caspase抑制剂Z-VAD-FMK处理受精后4天和5天的变态前幼虫,同时以抑制剂的有机溶剂DMSO(Dimethyl sulfoxide)作对照,根据其对幼虫变态的影响,分析Caspase在文蛤幼虫变态中的作用.细胞凋亡检测结果显示,幼虫发育过程中发生细胞凋亡的部位主要在壳顶幼虫的面盘,说明细胞凋亡可能是文蛤幼虫变态过程中面盘退化的主要机制;在担轮幼虫至壳顶幼虫中都检测到有活性的Caspase,说明整个幼虫形态变化过程中都有Caspase的参与;用Caspase特异性抑制剂处理变态前幼虫,发现处理组幼虫变态率均显著低于对照组(P<0.05),说明Caspase在变态早期和晚期都有重要作用,同时发现DMSO对幼虫变态有诱导作用,可以在生产中加以利用.     

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.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

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.     

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。     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

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.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.     

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.