红鳍东方鲀性腺的组织学分化

对中国土著红鳍东方鲀(Takifugu rubripes)种群人工培育苗种的性腺分化进行了组织学研究。结果表明:原始生殖细胞在孵化后2日龄(全长2.73mm)时迁移至中肾管和肠管之间;孵化后12日龄(全长3.59mm)原始性腺形成;孵化后40日龄(全长18.06mm)卵巢腔原基形成,标志着原始性腺开始向卵巢方向分化;至82日龄(全长55.42mm)卵原细胞进入第一次减数分裂时期,初级卵母细胞形成。原始性腺开始向精巢分化的时间较卵巢晚,孵化后61日龄(全长33.62mm)输精小管形成,标志着精巢分化的开始;孵化后103日龄(全长76.45mm)精原细胞进入减数分裂时期,初级精母细胞形成。研究结果为红鳍东方鲀性别控制和全雄苗种制种技术的研究奠定了基础。     

军曹鱼(Rachycentron canadum)性腺分化及首周年发育的组织学观察

本研究采用石蜡组织切片和H.E.染色法对军曹鱼原始性腺的形成、分化及精巢和卵巢首周年发育的组织结构变化进行观察。结果显示,军曹鱼原始生殖细胞在7孵化日龄(days post hatching,dph)时迁移到达生殖嵴,随后体细胞出现聚集和分裂,并于15 dph将原始生殖细胞完全包绕形成原始性腺。军曹鱼卵巢分化的时间要早于精巢,34 dph的稚鱼性腺组织尚未分化,但可观察到两种存在明显组织学差异的性腺类型,其中一种类型的性腺出现卵原细胞群,而另一种类型的性腺横切面较狭长,生殖细胞数量明显较少,因此推定为未分化精巢;卵巢的解剖学分化开始于44 dph,其标志为卵巢腔的形成;50 dph时精巢开始细胞学上的分化,此时精原细胞由基底膜包被形成囊泡状的细胞团。在首周年发育期间 (60～360 dph),军曹鱼的精巢发育包含I、II、III、IV、V 5个时期,而卵巢发育只包含I、II、III 3个时期。60 dph时,精巢和卵巢均处于I期;90 dph时,精巢发育至II期,卵巢仍处于I期;120 dph时,超过半数的精巢已发育至 III期,仅少部分卵巢发育至II期;150 dph时,精巢已发育至III期,而大部分卵巢发育至II期;185 dph时,精巢仍为III期,卵巢均已发育至II期;210 dph时,大部分精巢发育至IV期,卵巢仍处于II期;360 dph时,精巢已发育至V期,大部分卵巢发育至III期。上述研究结果可丰富军曹鱼的繁殖生物学研究基础,阐明其早期性腺发育规律,还可为其人工繁殖提供理论依据。     

犬齿牙鲆胚胎及仔、稚、幼鱼形态发育观察

犬齿牙鲆(Paralichthys dentatus)卵为端黄卵,受精卵浮性,卵径950~1100μm,油球径200~225μm.在水温20.5~22.0℃,盐度33条件下培育,受精后1 h 20 min 进入卵裂期,6 h 10 min 进入囊胚期,11 h 20 min 进入原肠胚期,19 h 10 min 发育到胚孔封闭期,胚体形成,26 h 50 min 尾芽出现,33 h 35 min心跳开始,48 h 10 min 开始孵化出仔鱼.初孵仔鱼平均全长2.59 mm±0.02 mm,在培育水温为17.0~21.0℃,盐度24~34条件下,胚后发育时序为:1日龄~9日龄为仔鱼前期,10日龄平均全长5.52 mm±0.05 mm,卵黄囊和油球完全吸收进入仔鱼后期;40日龄平均全长20.82 mm±0.67 mm,各鳍鳍条发育形成进入稚鱼期;65日龄平均全长41.13 mm±0.88 mm,全身鳞被完整进入幼鱼早期     

黑鲷早期鳞被的形成

黑鲷Sparus macrocephalus(Basilewsky)幼苗鳞被最早出现的部位是在躯干部胸鳍基后至肛门直上方的侧线处。黑鲷只具一个鳞被起始中心。最早出现鳞片的个体为体长9.5—14.1mm,平均11.51±0.85mm,全长11.2—16.5mm,平均13.65±1.01mm,孵后日龄23—35d。鳞被发育完全时的最小个体为体长12.1—18.5mm,平均14.57±1.55mm,全长14.8—23.5mm,平均17.99±2.17mm,孵后日龄27—42d。     

犬齿牙鲆早期形态特征和生长、发育规律

为掌握犬齿牙鲆早期发育的规律,2006年3-6月在实验室利用显微数码摄像系统,通过电脑观察、测定、拍摄犬齿牙鲆仔、稚、幼鱼各期的发育时间及早期的形态特征.犬齿牙鲆室内人工育苗的培育水温19～21 ℃,S为30,pH为8.2,光照度为500 lx左右,饵料依次为经过强化的轮虫、卤虫无节幼体和配合饲料.初孵仔鱼全长2.2～2.5 mm,身体透明,至4～5日龄,卵黄囊缩小,开始摄食,6～7日龄卵黄囊吸收完毕进入后仔鱼期;至29日龄全长9.0～9.4 mm,冠状幼鳍形成完善,开始进入稚鱼期;43日龄时全长15.0～16.0 mm稚鱼右眼完全迁移至左侧,变态完成,营底栖生活进入幼鱼期;幼鱼期鳞被形成完整,基本具有成鱼的外部形态和生活习性.     

漠斑牙鲆仔、稚、幼鱼的形态发育研究

2006年12月至2007年2月,在福建省漳州市东山县华涛水产综合养殖场,对漠斑牙鲆(Paralichthys lethostigma)仔、稚、幼鱼各发育阶段的形态特征进行了观察和描述.初孵仔鱼平均全长2.73±0.211mm.在培育水温18.1～23.0℃和盐度为33～34条件下,初孵仔鱼至孵化后7日龄为前期仔鱼,6日龄仔鱼开口;孵化后8日龄至30日龄为后期仔鱼;孵化后31日龄至45日龄为稚鱼期;孵化后46日龄进入幼鱼期.冠状幼鳍的消长、右眼的移位以及体色的左右不对称是漠斑牙鲆早期发育过程中最明显的外部形态变化.     

牙鲆sox9基因的克隆与表达研究

通过Genome walking方法克隆得到了牙鲆(Paralichthys olivaceus)sox9的基因组全长序列,对其在成鱼各组织的差异表达进行了RT-PCR分析.结果显示:牙鲆sox9基因全长3573 bp,包含3个外显子和2个内含子,编码的蛋白全长478 aa,含有高度保守的HMG结构域;sox9基因在性腺的表达具有两性差异,精巢中的表达明显强于卵巢中的表达,属1个性别相关基因,但其在肝脏、心脏、鳃、脑、眼和胃等多种组织中也有不同程度的表达.据此进一步分析和讨论等sox9基因在牙鲆性腺发育中的作用.     

牙鲆17β-HSD1基因克隆及其表达调控的初步研究

17β-羟类固醇脱氢酶1(17β-HSD1)的主要作用是将雌酮(El)转化为发挥功能的雌二醇(E2)。作者从牙鲆(Paralichthys olivaceus)性腺转录组数据库获得该基因的开放阅读框(ORF)序列,对其进行了验证,并分析了该基因在高温、外源性激素处理条件下性腺分化期性腺组织中的差异表达以及c AMP和转录因子(NR5a2和NR0b1)在精巢原代细胞中对该基因表达的作用。结果显示,牙鲆17β-HSD1基因的ORF为873bp,编码290个氨基酸,与其他鱼类的有很高的相似性。半定量RT-PCR结果表明,该基因在卵巢中高表达,精巢有少量表达,并且在雌性个体的鳃、头肾、肾、脾、胃和肠中也有表达。实时定量RT-PCR结果显示,该基因在卵巢或精巢分化的关键时期表达量较高;在精巢原代培养细胞中,外源信号分子c AMP及转录因子NR5a2可以显著下调17β-HSD1基因的表达(P0.05),且呈现剂量效应,转录因子NR0b1对该基因的调控也与剂量有关。作者推测牙鲆17β-HSD1基因在性腺分化中起一定的作用,其表达受到调控因子的作用,这些结果将有助于增加对鱼类性腺分化和发育的认识。     

已烯雌酚诱导黑鲷幼鱼性转换

黑鲷Sparusmacrocephalus是雌、雄同体,雄性先成熟鱼类。在人工养殖条件下,2龄鱼全为雄性,3龄鱼个别转为雌性。本项研究的目的是使用性激素促使黑鲷提前转为雌性,缩短产卵周期。本次实验是使用当年幼鱼,目的在于进一步找出较佳的处理年龄和使用的激素剂量。1　材料与方法1.1　材料实验使用的黑鲷为1994年孵出,至同年11月16日实验开始时,鱼龄为6个月。这批实验用鱼取35尾测量,全长范围为155～185mm,平均169mm;体长131～154mm,平均139mm;体重67～112g,平均94g。其中10尾鱼取性腺做组织切片观察,有60%已分化出卵巢和精巢,其余未分化。1.2　方…     

罗氏沼虾(Macrobrachium rosenbergii)初次性成熟性腺发育的组织学观察

性腺发育是罗氏沼虾(Macrobrachium rosenbergii)个体生长及繁育的重要生物学过程,是良种培育的基础。采用外部形态观察及常规石蜡切片的方法,分析了罗氏沼虾初次性成熟过程中性腺外部形态及内部组织学变化的规律。结果表明,性腺颜色、体积及细胞组成、分布均随性腺发育表现出规律性变化,卵巢发育划分为8个时期,精巢发育分为4个时期。卵巢外部形态发育与内部组织学变化同步性高,随卵巢成熟度增加,不同阶段生殖细胞由生发区向外形成环状分区, Ⅰ~Ⅲ期卵巢体积、生殖细胞大小及性腺指数(GIS)增长缓慢,主要进行生殖细胞数量积累, Ⅳ~Ⅶ期卵巢体积、生殖细胞大小及GIS增长迅速,生殖细胞在增殖同时,营养物质积累迅速。可根据卵巢颜色、体积、色素分布等推测其内部组织细胞的发育情况。精巢形态发育和组织发育同步性较差,精巢体积、生殖细胞大小、种类及GIS在精巢成熟过程中变化较小, Ⅰ期精巢主要进行个体生殖功能的完善, Ⅱ期、Ⅲ期、Ⅳ期精巢则整体表现为个体生殖能力的强化过程。研究结果丰富了罗氏沼虾繁殖生物学内容,从形态及组织学方面初步探明了罗氏沼虾性腺的基本发育过程,可为其良种选育和虾苗繁育提供参考依据。     

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.     

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.     

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。     

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.     

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.     

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.