一株海绵附生菌Bacillus firmus的培养条件优化及α-GI活性成分初步分析

为寻找新型α-葡萄糖苷酶抑制(α-glucosidase inhibition,α-GI)活性化合物来源,针对前期研究发现的具有α-GI活性的海绵附生微生物坚强芽孢杆菌(Bacillus firmus),对其发酵培养条件进行优化,并对其活性组分中主要成分进行初步分析.以α-GI活性为主要检测对象,选出蛋白胨、酵母膏、Na Cl和p H值共4个因素进行正交优化试验.结合菌株生长情况和代谢产物产量指标,发现在MB培养基的基础上,蛋白胨质量浓度3 g/dm3、酵母膏质量浓度3 g/dm3、Na Cl质量浓度19.5 g/dm3、p H值为7.5的组合为最优培养条件.在优化后的培养条件下,规模化发酵该菌株,提取其代谢产物粗提物,经活性追踪下的二次色谱柱分离,获得一个高抑制活性组分,抑制率达82.4%.该组分经GC-MS分析后发现,活性组分中主要成分为环二肽类及哌啶酮类物质.本研究结果为寻找新型α-GI活性化合物提供了一个新的、可扩大生产的来源.     

一株海绵附生芽孢杆菌的抗硅藻附着活性成分的分离鉴定

为寻找天然抗污损活性化合物,以抗硅藻附着活性为导向,采用有机溶剂萃取、半制备高压液相色谱对分离自海绵的芽孢杆菌UST050418-715代谢产物进行分离,纯化抗硅藻附着活性物质,并利用气相色谱-质谱联用仪、核磁共振波谱分析活性物质结构。从菌株UST050418-715代谢产物中分离得到7种具有抗硅藻附着活性的环二肽类化合物,分别鉴定为:(1)环(L-亮氨酸-反式-8-羟基-L-脯氨酸-)、(2)环(L-缬氨酸-L-脯氨酸)、(3)环(D-脯氨酸-L-亮氨酸)、(4)环(L-脯氨酸-D-亮氨酸)、(5)环(甘氨酸-L-脯氨酸)、(6)环(L-苯丙氨酸-顺式-8-羟基-D-脯氨酸-)、(7)环(L-苯丙氨酸-反式-8-羟基-L-脯氨酸-)。说明海绵附生芽孢杆菌UST050418-715代谢产物中存在大量环二肽类化合物,可以帮助宿主海绵实现对硅藻附着的化学防御。     

抗硅藻附着活性细菌的筛选、鉴定与发酵条件优化

从美国华盛顿州圣璜岛海域的8 种海绵样品的120 株共附生微生物中, 筛选出11 株细菌的粗提物能有效抑制硅藻附着。其中683 号菌株活性最高, 对5 种受试硅藻的平均抑制率达到98.0%。结合形态学特征和扫描电镜观察, 以及16S rDNA 序列分析, 鉴定该菌株为短小芽孢杆菌Bacillus pumilus。并采用正交试验对该菌株进行发酵条件优化, 结合粗提物的产量和抗硅藻附着活性进行统计学分析,确定了该菌株的最佳发酵条件是: 发酵温度25℃, pH 7.5, NaCl 浓度19.45 g/L, 蛋白胨浓度5 g/L。     

海水中铬(Ⅵ)对浮游植物生长率的影响

本工作使用一次培养法初步研究了Cr(Ⅵ)对菱形硅藻和隐藻的毒性,实验结果表明,当Cr(Ⅵ)浓度大于16 ppb时,则对其生长率产生抑制作用,其72小时EC₅₀值分别为260ppb和230ppb.根据活性点位模式计算了Cr(Ⅵ)与藻体的结合常数,约为2×105.藻体中Cr含量与培养液中Cr浓度有关,两者近似指数关系,根据藻体中Cr含量和溶液中Cr的浓度,估计每个硅藻细胞与Cr最大结合容量约为5×1017mol/dm3.     

热稳定k-卡拉胶酶产生菌Bacillus sp.Car19的发酵条件优化

对一株分泌热稳定κ-卡拉胶酶印尼热泉菌进行了种属鉴定,并采用响应面法对该菌发酵产酶条件进行了优化。鉴定结果表明,该菌株属于芽孢杆菌属(Bacillus),命名为Bacillus sp.Car19(Gene Bank:KT865196)。发酵条件优化结果显示,9个环境因子影响Bacillus sp.Car19产酶量。其中影响Bacillus sp.Car19产酶量的三个主要因素分别为培养温度、培养基中Cu~(2+)浓度和培养基中Na Cl浓度。综合次要因素对Bacillus sp.Car19产酶影响,Bacillus sp.Car19最佳产酶发酵条件为:培养温度52.31℃、Cu~(2+)浓度6.93 mmol/L、Na Cl浓度37.03 g/L,培养基p H为6,接种量1%,培养时间36 h,半乳糖浓度0.3 g/L,硝酸铵浓度7 g/L,卡拉胶浓度0.5 g/L。优化后发酵上清液酶活力达到15.21 U/m L,与优化前相比提高了1.5倍。     

一株海洋微藻的分离鉴定及其产油条件优化

自青岛近岸海水中分离到一株硅藻,经形态学和分子生物学鉴定,表明该藻与小头菱形藻(Nitzschia microcephala)最为相近,18SrDNA相似性99%以上。研究以f/2培养基为基础,对该藻的生长和油脂积累条件进行探析,表明其在pH=6~8时具有较高的生长速率,最大可达到0.01d-1,油脂含量随pH的升高而降低;添加SiO2-3可以抑制菱形藻的生长和油脂积累;高浓度的Fe3+能促进菱形藻的生长;在Zn2+浓度为1.74μmol/L的条件下,菱形藻能够较好地生长并且进行油脂的积累;其中pH、SiO2-3和Fe3+是影响该藻生长和油脂积累的主要因子。正交试验研究表明,该藻在f/2培养液中生长和油脂积累的最优条件为初始pH=9,SiO2-3和Fe3+添加浓度分别为0.07和100μmol/L。在该最优条件下,该藻的生物量和油脂积累可分别达到94.7mg/L和19.6%。     

一株高效褐藻酸降解菌的筛选、鉴定及其发酵条件的优化

以褐藻酸钠为唯一碳源,从腐烂海带中筛选得到褐藻酸钠降解能力较强的菌株H4,经生理生化和16S rDNA序列分析鉴定该菌株属于交替单胞菌属(Alteromonas)。对菌株H4的发酵条件优化研究表明,H4的较优培养基组成为(w/v):褐藻酸钠0.6%、酵母粉0.5%、蛋白胨0.25%、NaCl 3%;较优培养条件为:培养温度28℃,初始pH7.5,接种量1.5%(v/v),培养时间48 h。Fe3+对交替单胞菌H4的产酶有明显的促进作用,这在其他褐藻酸裂解酶生产菌株中未见报道,而Cu2+对褐藻酸裂解酶的抑制作用高达45.78%。在优化后的培养条件下,粗酶液酶活达到146.45 U/mL,较优化前提高了39.4%。     

三种海洋微藻叶绿素a含量对抗生素胁迫的响应变化

实验室条件下,研究不同抗生素胁迫对小球藻(Chlorella vulgarisBeij.)、金藻8701(Isochrysis galbana Parke8701.)和小新月菱形藻(Nitzschia closteriumEhr.)生长的影响。结果表明:1)低于100μg.mL-1的氯霉素作用下,小球藻和金藻8701叶绿素a含量变化与对照组无明显差异;小新月菱形藻叶绿素a含量显著降低。2)不同浓度梯度的遗传霉素(G418),均显著抑制3种微藻叶绿素a含量的增加。3)青霉素浓度低于100μg.mL-1时,能够促进3种藻叶绿素a的增加,但不同微藻间叶绿素a含量的相对增长率存在明显差异。试验结果可为微藻的基因工程选择标记和无菌培养体系的建立提供参考。     

3种海洋硅藻不同培养阶段挥发性成分的比较分析

本文应用顶空固相微萃取气相色谱-质谱联用技术,对威氏海链藻Thalassiosira weissflogii、小新月菱形藻Nitzschia closterium、角毛藻Chaetoceros calcitrons 3种硅藻的挥发性成分进行分析。结果表明,在3种硅藻的培养过程中,其挥发性成分除了壬醛及醇类呈现先增后降的趋势外,大多数不饱和醛类的含量在不断增加,烷烯烃类物质在不断减少,而乙酸乙酯、2-甲基-2-丙酸甲酯这类酯类化合物在角毛藻衰败期会显著富集。通过对代表性饵料硅藻的挥发性成分进行研究,可为海水养殖过程中微藻投喂种类、投喂时机的选择提供理论依据,并为海洋硅藻的进一步开发奠定基础。     

大亚湾拟菱形藻水华及其在生物群落中的生态地位

对2000年4~5月藻类水华高发期大亚湾澳头海域的拟菱形藻进行逐日调查,并对其与浮游植物、浮游动物的相关关系进行分析.调查初期,整个生物群落的种类组成数量都相对贫乏;拟菱形藻数量急剧增长,最高的测站密度可达2.93×106个/dm3.拟菱形藻水华的组成种类有拟极细拟菱形藻和尖刺拟菱形藻两种,以尖刺拟菱形藻稍占优势.水华形成中其他种类硅藻和甲藻在种类组成和数量上也有一定程度的增加.分析表明,拟菱形藻在浮游植物群落的种间竞争中具有优势,并成为春季硅藻水华的最重要的组成者.桡足类等浮游动物对拟菱形藻的下行控制并不能明显抑制拟菱形藻的增长及水华的产生,这与其偏爱以其他种类藻类为食有关.以上因素外加拟菱形藻对环境的广泛适应性促使其在该水域成为最常见的赤潮生物之一.     

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。     

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.     

《Acta Oceanologica Sinica》Vol. 33(2014) contents

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INFORMATION FOR AUTHORS

<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,