两株芽孢杆菌对凡纳滨对虾生长和血清非特异性免疫指标的影响研究

为研究芽孢杆菌对凡纳滨对虾(Litopenaeus vannamei)生长指标和血清非特异性免疫指标的影响。以初始体重(4.00±0.03)g的凡纳滨对虾为对象,在对虾饲料中分别添加浓度为1×10~7、1×10~9和1×10~(11)cfu/kg的蜡样芽孢杆菌(Bacillus cereus)和枯草芽孢杆菌(B.subtilis),以对虾商品饲料为对照组,养殖42d。实验结束后,测定凡纳滨对虾的生长情况和血清非特异性免疫指标。研究表明:各组对虾的成活率均在90%以上,且组间差异不显著(P0.05);除了枯草芽孢杆菌1×107cfu/kg添加组,其余添加组中凡纳滨对虾的体重、特定生长率、饲料效率和消化率(P0.05)显著提高,在添加浓度为1×109cfu/kg时各指标取得最大值。饲料中添加益生菌可提高凡纳滨对虾的超氧化物歧化酶、碱性磷酸酶、酸性磷酸酶、溶菌酶、总一氧化氮合酶、诱导型一氧化氮合酶活性和血清蛋白浓度等血清非特异性免疫指标,且均在添加浓度为1×109cfu/kg时各指标取得最大值。研究结果表明,饲料中添加适当浓度的益生菌可有效促进凡纳滨对虾的生长,提高其血清非特异性免疫水平。当益生菌的添加浓度为1×109cfu/kg时,效果最显著。     

抗生素和有益微生物对凡纳滨对非特异性免疫效应的研究

凡纳滨对虾为实验对象,进行了抗生素和有益茵对对虾免疫效应的研究.抗生素组每天泼洒氯霉素1次,浓度约为2mg/dm3;有益茵组每隔7d泼洒1次有益微生物,包括利生活茵90g和EM18cm3.实验为期1个月,每10d取样1次,检测凡纳滨对虾血清、肌肉和肝脏中的溶茵力、抗茵力、酚氧化酶、碱性磷酸酶、超氧化物歧化酶.结果表明,除个别情况外,抗生素组的凡纳滨对虾血清、肌肉和肝脏中的溶茵力、抗茵力、酚氧化酶、碱性磷酸酶、超氧化物歧化酶低于对照组,而有益茵组则高于对照组.由此可以推断,抗生素对凡纳滨对虾非特异性免疫系统具有一定的抑制作用,而有益茵则可以从一定程度上提高凡纳滨对虾非特异性免疫水平.     

高位池养殖对虾携带白斑综合症病毒变化

采用实时荧光定量 PCR 技术研究6口高位池塘斑节对虾和凡纳滨对虾携带WSSV变化,结果表明:养殖过程中凡纳滨对虾WSSV携带量最高为9.7×105拷贝/g;斑节对虾最高携带量为9.5×105拷贝/g.凡纳滨对虾WSSV感染率分别为:苗种没有携带WSSV;30d为80.0%;60 d为90.O%;90 d为90.0%;120 d为93.3%,斑节对虾潜伏感染率分别为:苗种没有携带WSSV;30d为73.3%;60d为83.3%;90d为90.O%:120d为96.7%.     

二溴海因和碳水化合物水平对凡纳滨对虾生长和免疫的影响

研究了低盐度条件下,二溴海因和碳水化合物水平对凡纳滨对虾(Litopenaeus vannamei)生长和免疫的影响,共3个实验。实验1研究了二溴海因对凡纳滨对虾存活的影响,发现在0.2、5和20三个盐度水平,随着二溴海因浓度的升高,凡纳滨对虾存活率呈下降趋势;二溴海因对凡纳滨对虾的安全浓度随盐度的增加呈上升趋势。实验2和实验3研究了盐度、二溴海因浓度和饲料中碳水化合物水平对凡纳滨对虾生长和免疫的影响,结果表明:在低盐度养殖条件下,凡纳滨对虾长期生活于二溴海因环境中,可导致对虾处于应激状态,需要消耗体内较多的免疫因子,血清酚氧化酶和超氧化物歧化酶活性降低,生长速度下降,而在饲料中适量增加碳水化合物不仅可促进对虾生长又可起到饲料蛋白质节约作用。     

副溶血弧菌、脂多糖和嗜酸小球菌对凡纳滨对虾血清一氧化氮合酶的影响

探讨了凡纳滨对虾Litopenaeus vannamei血细胞中是否存在一氧化氮合酶(NOS)活性,进行了副溶血弧菌Vibrioparahaemolyticus和哈维氏弧菌Vibrioharveyi脂多糖(LPS)体外孵育实验,采用亚硝酸盐法对凡纳滨对虾血细胞中NOS活性进行了测定。结果表明,副溶血弧菌和LPS离体孵育血细胞组的一氧化氮(NO)浓度显著高于对照组,而且这一反应能被NOS抑制剂亚硝基L精氨酸甲酯(LNAME)所阻断,证实了凡纳滨对虾血细胞中存在NOS活性。同时进行了副溶血弧菌感染凡纳滨对虾对其血清NO浓度和NOS活性的影响试验。试验表明,副溶血弧菌注射感染凡纳滨对虾后12h血清中NOS活性极显著高于对照组;NO浓度在注射后24h开始升高,72h后各组NO浓度都显著高于对照组。说明副溶血弧菌感染凡纳滨对虾可以诱导NOS的表达,并产生NO来抵抗副溶血弧菌的入侵,从而增强了对虾的免疫力。此外将嗜酸小球菌Pediococcusacidilactici按10.0mg·kg-1添加到饲料里投喂凡纳滨对虾后,血清中溶菌酶活力和NOS活性比对照组有显著提高,且溶菌酶活力和NOS活性存在显著正相关性(r=0.629,r0.05[1,12]=0.532)。从而进一步说明了NOS活性可以作为凡纳滨对虾免疫力高低的有效指标。     

凡纳滨对虾遗传多样性研究中AFLP体系的建立

凡纳滨对虾是当今世界公认的重要对虾养殖品种之一,然而良种选育研究仍相对落后,亟需采用分子遗传学手段加以解决.本文以凡纳滨对虾为材料,对预扩增体系、选择性扩增体系中的稀释倍数、dNTP浓度、Taq浓度、Mg2+浓度、引物浓度进行比较,结果表明,300 ng基因组DNA采用5U PstI和5UMseI双酶切4 h,16℃连接过夜,连接产物稀释10倍进行预扩增,预扩增产物稀释50倍进行选择性扩增,能够得到稳定丰富的条带,最终建立了适用于凡纳滨对虾的AFLP反应体系.本研究又进一步应用优化后的AFLP反应体系,从64对选择性扩增引物组合中筛选出12对多态性高的引物组合,便于对凡纳滨对虾基因多态性位点进行分析,从而为凡纳滨对虾的遗传多样性研究奠定基础.     

不同盐度下凡纳滨对虾血淋巴免疫生理指标比较

凡纳滨对虾（Litopenaeus vannamei）在不同盐度（4，18，32）和温度（22-26℃）水体中养殖，分别于1，5，15和30d后取样，进行不同盐度下凡纳滨对虾血淋巴免疫生理指标比较。结果表明：盐度对凡纳滨对虾血淋巴酚氧化酶、超氧化物歧化酶、酸性磷酸酶和碱性磷酸酶活力均有显著影响（P〈0．05）。盐度由32渐变至设定盐度后，实验前期（15d之内），盐度4和18组凡纳滨对虾上述各项指标均呈现一定的波动趋势，至15d后趋于稳定；盐度32组在整个实验过程中基本稳定。稳定后的各项指标均表现在盐度18时最高，盐度4时次之，盐度32时最低。     

凡纳滨对虾在盐度变化下免疫机能评价的研究

研究凡纳滨对虾(Litopenaeus vannamei)在盐度变化下免疫机能的评价指标。结果表明:盐度变化对凡纳滨对虾血细胞数量、吞噬率和血淋巴酚氧化酶、溶菌、α2-巨球蛋白活力的影响显著(P<0.05),而对照组各免疫指标无明显变化。凡纳滨对虾在盐度变化1d内3类血细胞和总血细胞数量明显下降,1d后趋于稳定,且与盐度值呈正相关;血细胞吞噬率和血淋巴酚氧化酶、溶菌和类α2-巨球蛋白活力在3d内呈峰值变化,其中血细胞吞噬率、溶菌活力均于0.5d时达到最小值,血淋巴酚氧化酶、类α2-巨球蛋白活力分别于0.5d或1d时达到最大值,同时各免疫指标与盐度变化呈明显的正、负相关性,3d后均恢复至对照组水平。结合凡纳滨对虾在盐度变化下的免疫响应机制和免疫适应性,提出血细胞吞噬率和血淋巴溶菌活力可作为衡量对虾在盐度变化下免疫健康水平的指标。     

三种益生菌及其复合菌对凡纳滨对虾生长和血清非特异性免疫的影响

为研究3种益生菌及其复合菌对凡纳滨对虾(Litopenaeus vannamei)的生长和血清非特异性免疫的影响。以基础饲料为对照组,在基础饲料中分别添加浓度为1×109cfu/kg的蜡样芽孢杆菌(Bacillus cereus)、嗜酸乳杆菌(Lactobacillus acidophilus)和酪酸菌(Clostridium butyricum),并以1∶1∶1比例添加浓度为1×109和3×109cfu/kg的复合菌,共5个处理组。养殖42d后,测量了5组凡纳滨对虾的生长与成活率、饲料效率、特定生长率和超氧化物歧化酶、溶菌酶、碱性磷酸酶及酸性磷酸酶的活性。研究表明:凡纳滨对虾的成活率均在90%以上,各处理间差异不显著(P0.05)。饲料中添加3种菌及其复合菌均可显著提高凡纳滨对虾的体重、特定生长率和饲料效率等指标,且均在酪酸菌组取得最大值并显著高于其它处理组(P0.05)。饲料中添加3种益生菌及其复合菌均能不同程度地提高凡纳滨对虾的超氧化物歧化酶、溶菌酶、碱性磷酸酶和酸性磷酸酶等血清非特异性免疫指标,各处理组均显著高于对照组(P0.05),其中酪酸菌组各免疫指标均取得最大值。研究结果显示,饲料中添加3种菌及其复合菌均可有效促进凡纳滨对虾的生长,提高其血清非特异性免疫水平,其中以添加酪酸菌的效果最为显著。     

循环水养殖系统在凡纳滨对虾种虾养殖中的应用效果初探

循环水养殖系统在凡纳滨对虾种虾养殖中应用较少,本研究应用循环水养殖系统养殖凡纳滨对虾种虾,设定4个不同的养殖密度(30、40、50、65尾/m2),初始体重:(0.102±0.008)g,研究凡纳滨对虾种虾在循环水养殖系统中的生长情况。养殖期间定时对对虾体重和水体指标(氨氮、亚硝酸氮、pH、水温、微生物)进行分析测定。通过对各项数据分析表明:低密度组(30、40、50尾/m2)凡纳滨对虾体重增长较快,各组特定生长率分别为(3.83±0.03)%、(3.87±0.01)%、(3.81±0.03)%,绝对增重率分别为(0.201±0.009)、(0.214±0.004)、(0.194±0.009)g/d,但均无显著性差异(P0.05);高密度组(65尾/m2)的凡纳滨对虾体重增长较慢,特定生长率和绝对增重率分别为(3.41±0.02)%和(0.107±0.004)g/d,该结果与低密度组间存在显著性差异(P0.05)。低密度组中凡纳滨对虾养殖水体的水质指标要优于高密度组,4个密度组中氨氮、亚硝酸氮、绿菌均维持在安全浓度范围内,仅仅黄菌数量略高。综合分析,采用该养殖系统养殖凡纳滨对虾的最优密度为50尾/m2。因此,本研究可为循环水养殖系统养殖凡纳滨对虾种虾提供参考。     

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.     

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.     

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.