一株溶藻菌Zobellella sp. B307对太平洋亚历山大藻的溶藻特性及作用机制研究

为研究溶藻菌对赤潮甲藻的作用机制,本文从胶州湾沉积物中分离得到的一株耐盐菌株Zobellella sp. B307,并研究了该菌株对太平洋亚历山大藻(Alexandrium pacificum)的抑制效果,从细胞结构、生理水平和分子水平探究了该菌株的抑藻途径及作用机制。结果表明,该菌株的抑藻途径为分泌胞外活性物质的间接溶藻,72 h的溶藻率高达91%。溶藻过程中藻细胞壁破损,叶绿素a和总蛋白含量显著下降,藻细胞的活性氧(ROS)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性显著高于对照组,这表明溶藻物质对藻细胞造成的强烈氧化损伤是导致其死亡的直接原因。热休克蛋白基因(HSP)表达量显著上调,说明溶藻物质激发藻细胞产生的ROS在激活抗氧化系统的同时诱导产生HSP70,二者联合清除ROS以减缓藻细胞受应激损伤的程度;A.pacificum的网格蛋白基因(Clathrin)表达量显著上调,表示由网格蛋白介导的胞吞作用明显增强,进而说明为减轻光合营养功能受损造成的影响,藻细胞的异养功能增强。本研究结果有助于诠释菌藻关系,更为探究赤潮治理的生物方法提供重要的理论支持和实践指导。     

三角褐指藻与东海原甲藻间的他感作用研究

本文主要探究了东海原甲藻(Prorocentrum donghaiense)与三角褐指藻(Phaeodactylum tricornutum)之间的他感作用。在实验室模拟条件下,分别研究了添加比例为10%、40%、60%、90%、100%的三角褐指藻无藻细胞滤液的培养基对东海原甲藻生长的影响,以及相同比例的东海原甲藻无藻细胞滤液培养基对三角褐指藻生长的影响。结果表明,低浓度三角褐指藻滤液对东海原甲藻(P.donghaiense)的生长表现为促进作用,高浓度则抑制其生长,因此,不同浓度的三角褐指藻藻液对东海原甲藻生长表现出两种相反的他感作用效果;而东海原甲藻滤液对三角褐指藻(P.tricornutum)的生长没有显著影响,说明东海原甲藻并未对三角褐指藻表现出他感作用。     

溶藻细菌MS7的分类鉴定及其溶藻活性的初步研究

采用形态学、生理生化指标和16SrRNA基因序列同源性分析的方法,对菌株MS7进行鉴定,并通过溶藻试验测定该菌株对铜绿微囊藻(Microcystis aeruginosa)的溶藻活性和溶藻方式。结果表明,MS7菌株为苏云金芽孢杆菌(Bacillus thuringiensis),对铜绿微囊藻具有高效溶藻效果,当菌浓度为108cells/mL,初始藻液叶绿素a浓度为0.7mg/L时,48h溶藻率达到91%。主要通过分泌胞外物质的间接方式溶藻,溶藻物质属于具有很强的热稳定性的非蛋白质类物质。     

剧毒卡尔藻与东海原甲藻间的化感作用研究

为研究剧毒卡尔藻(Karlodinium veneficum)与东海原甲藻(Prorocentrum donghaiense)之间的化感作用,在f/2营养盐条件下,分别采用了混合培养及滤液培养的方法。结果表明:剧毒卡尔藻指数生长后期的无藻细胞滤液明显抑制了东海原甲藻的生长,而东海原甲藻滤液对剧毒卡尔藻的生长几乎无影响,化感作用作为重要的影响因素使剧毒卡尔藻获得竞争优势。将剧毒卡尔藻指数生长末期的无藻细胞滤液置于固相萃取(SPE) C18小柱,经吸附、不同比例的甲醇溶液洗脱,考察不同洗脱部分对东海原甲藻生长的影响,发现80%的甲醇洗脱部分明显抑制了东海原甲藻的生长。本研究为了解剧毒卡尔藻的竞争机制提供了重要的科学依据,今后将重点分析剧毒卡尔藻释放的有效抑制东海原甲藻生长的化感物质。     

网状原角藻化感作用的初步研究

为明确海洋甲藻——网状原角藻(Protoceratium reticulatum)对海洋微藻的化感作用,揭示其化感物质作为植物源抑藻剂的开发潜能,本文探讨了网状原角藻藻细胞和无细胞滤液对几种海洋微藻——东海原甲藻(Prorocentrum donghaiense)、拟旋链角毛藻(Chaetoceros pseudocurvisetus)、米氏凯伦藻(Karenia mikimotoi)和赤潮异弯藻(Heterosigma akashiwo)生长的影响,并进一步选取东海原甲藻和赤潮异弯藻作为受体藻种,以塔玛亚历山大藻(Alexandrium tamarense)作为阳性对照,比较了2种甲藻的藻细胞培养液和无细胞滤液对东海原甲藻、赤潮异弯藻生长的影响差异。通过比较发现,网状原角藻的无细胞滤液比塔玛亚历山大藻能更明显的抑制东海原甲藻和赤潮异弯藻的生长,尤其是对东海原甲藻,基本阻断其生长。     

伊姆裸甲藻溶藻菌LD-B3的分离鉴定及溶藻作用

近年来,有害藻华频繁发生给海洋生态系统甚至人类健康带来严重影响。伊姆裸甲藻（Gymnodinium impudicum）是一种典型有害藻华原因种,开展其溶藻菌筛选工作,将有利于该种藻华的防控研究。本研究从伊姆裸甲藻藻华发生海域,分离一株针对藻华原因种的溶藻菌LD B3,16S rDNA序列分析表明,该菌株属于假交替单胞菌属（Pseudoalteromonas）。菌藻共培养结果显示,2.0%终浓度LD B3菌液添加72 h后,伊姆裸甲藻溶藻率达81.06%,且溶藻效应呈现浓度和时间依赖性。显微观察和生理参数测定结果表明,菌株LD B3可导致伊姆裸甲藻细胞链断裂,最大光合效率（Fv/Fm）下降。溶藻方式研究发现,菌株LD B3主要通过分泌胞外物质实现溶藻,且这类物质对温度较为敏感,初步推断其可能为蛋白类物质。本研究结果表明菌株LD B3在伊姆裸甲藻溶藻剂开发方面有一定应用前景。     

侧孢短芽孢杆菌溶藻活性代谢产物部分特性的研究

研究了不同因素对侧孢短芽孢杆菌（Brevibaci Uuslaterosporus）无菌滤液溶藻效果的影响,并对无菌滤液进行了紫外光谱扫描分析,探讨侧孢短芽孢杆菌溶藻活性代谢产物的物质特性、探究物质的组成成分．结果显示：无菌滤液分别经不同温度、pH值、蛋白酶和紫外照射时间处理后,均仍具有极为显著的溶藻能力（P〈0．01）,溶藻实验7d后颤藻（Oscillatoria sp．）叶绿素a质量浓度的平均下降率分别为：58．65％、63．88％、61．50％和62．61％,藻体干重的平均下降率分别为：39．77％、39．30％、36．49％和37．02％,表明溶藻活性代谢物质具有耐酸、耐碱、耐高温、抗蛋白酶水解、抗紫外辐射的特性;同时,分别在4、28℃环境中贮存28d的无菌滤液,始终保持着极高的溶藻活性,溶藻实验7d后颤藻叶绿素a质量浓度的平均下降率分别为：50．84％和60．39％,藻体干重的平均下降率分别为：37．82％和37．54％,说明该物质的结构极其稳定,在环境中可长久存在;另外,加热与未加热处理的无菌滤液的紫外光谱最大吸收峰都在290～300nm之间．分析表明,溶藻活性代谢产物极大程度上属于具有极强稳定性的肽类物质．     

磷酸盐对两种东海典型赤潮藻影响的围隔实验

采用了围隔生态实验方法,研究了围隔中营养盐含量及结构变化和磷酸盐对东海典型赤潮藻种(东海原甲藻和中肋骨条藻)生长的影响。结果表明,在高磷条件下,中肋骨条藻快速增殖,稳定期缩短,细胞数很快下降,而东海原甲藻由于氮源的限制生长期明显缩短。在磷限制条件下,中肋骨条藻细胞增殖受到明显抑制,生物量偏低,而东海原甲藻受到的影响较小。磷可能是东海原甲藻及中肋骨条藻生长的限制因子。氮磷比会影响浮游生物的生长,较高的氮磷比可能对东海原甲藻的生长有利,较低氮磷比则可能有利于中肋骨条藻生长。东海原甲藻能在营养盐浓度较低的生长环境中占有优势,相反,营养盐浓度较高的环境更适合中肋骨条藻的生长,中肋骨条藻具有更强的竞争能力。     

聚苯乙烯微塑料对东海原甲藻生长的影响

微塑料是海洋中的一种污染物,近年来已成为全球关注的热点环境问题。东海原甲藻（Prorocentrum donghaiense）是我国东海海域常见的有害赤潮藻,目前尚未有关于微塑料污染对东海原甲藻影响研究的报道。为了探讨聚苯乙烯（PS）微塑料对东海原甲藻生长的影响机制,阐明海洋微塑料对东海原甲藻生长的影响,本研究开展了东海原甲藻在聚苯乙烯微塑料差异化暴露条件下的生长动力学室内模拟实验。结果显示：①5 mg/L溶解性有机质（DOM）可促进东海原甲藻的生长;②10 mg/L（0.1 μm粒径）的聚苯乙烯微塑料对东海原甲藻前期生长的影响,表现为先抑制、后促进;③聚苯乙烯微塑料会抑制处于稳定期和衰退期东海原甲藻的生长;微塑料粒径越小、浓度越高抑制作用越强;浓度的抑制效应强于粒径的。我们研究发现聚苯乙烯微塑料会影响东海原甲藻的生长,这将为微塑料和赤潮藻复合污染生态效应以及海洋生态系统保护提供新的科学依据。     

东海原甲藻对中肋骨条藻的他感作用初探

在实验室内将不同生长期的东海原甲藻(Prorocentrum donghaienseLu)培养液滤液用切向超滤技术分级处理后,培养中肋骨条藻(Skeletonema costatum),发现东海原甲藻的滤液对中肋骨条藻的生长具有促进作用,不同生长期的东海原甲藻分泌物在粒级100 ku～0.45μm的培养液中对中肋骨条藻的促进作用最明显,表明东海原甲藻可以产生大小不同的可促进中肋骨条藻生长的他感物质,且在100 ku～0.45μm范围含量较大。东海原甲藻对中肋骨条藻的生长促进作用在不同生长期作用强度不同,促进作用最明显的是消亡期,其次为平台期,指数生长期的藻液是3种藻液中促进作用最小的。     

海洋生物共栖细菌抑藻活性的初步研究

针对我国常见赤潮藻种赤潮异弯藻(Heterosigma akashiwo)及海洋原甲藻(Prorocentrum micans),通过细胞活体荧光测定方法,研究了15株海洋生物共栖细菌对微藻生长的抑制作用,同时,对它们进行了分子生物学鉴定,分析了其系统发生学关系。结果表明,7株细菌的代谢产物对2种微藻的生长具有不同程度的抑制作用,并表现出一定的选择性,其中3株海洋细菌NJ6-3-2,QD1-2,ATCI02-4的代谢产物对2种赤潮微藻的生长都具有抑制作用。系统发生学分析显示,具有抑藻活性的海洋细菌中有4株(NJ6-3-1,QD1-2,SSN-6,SS6-4)属于-γproteobac-teria分支,另外3株(NJ5-7-1,NJ6-3-2,ATCI02-4)属于Gram 细菌。     

Algicidal activities of secondary metabolites of marine macroalgal-derived endophytic fungi

Red tide occurrs frequently and causes signi?cant damage to the environment and human health. As a result, development of new effcient and environment friendly red-tide microalgae inhibitors has gained increasing attention in recent times. Algicolous endophytic fungi with unique habitats are promising sources for active agents owing to their abundant secondary metabolites and distinguished activities. In this study, the algicidal activities of 49 marine macroalgal-derived endophytic fungi against phytoplankton Alexandrium tamarense, Prorocentrum donghaiense, Heterosigma akashiwo, and Chattonella marina were examined using 96-well microplate. Four fungal strains, including Aspergillus wentii(pt-1), A. ustus(cf-42),and A. versicolor(dl-29, pt-20), exhibited potent algicidal activities. A total of 32 pure compounds isolated from these fungi were noted to possess dif ferent degrees of algicidal activities. Of those, 11 compounds comprising ?ve anthraquinones, two terpenoids, and four steroids showed high 24-h inhibition rates for the four red tide algae, with 24 h EC₅₀ values ranging from 0.01 to 14.29 μg/mL. Among them, compound1(1,5-dihydroxy-3-methoxy-7-methylanthraquinone) presented the strongest activity against H. akashiwo,and could decrease its chlorophyll a(Chl a) and superoxide dismutase contents and increase the soluble protein, malondialdehyde, and peroxidase contents. These results suggested that the identi?ed anti-algal compound might inhibit the growth of red tide algae by weakening photosynthesis(reducing Chl a content),destroying cell membrane, and damaging the antioxidant system.     

微生物治理有害藻华研究进展

近年来,全球近海海域有害藻华频发,导致严重的环境问题与经济损失,如何有效地治理有害藻华引发全球关注。抑藻微生物可以通过细胞接触等直接方式及分泌胞外活性物质等间接方式破坏藻细胞的细胞结构、生理过程和遗传功能,进而抑制藻细胞的生长。本文总结了抑藻微生物控制有害藻华的主要作用原理和研究进展,综述了当前以微生物为基础的控制藻华的方法和技术,探讨了其在有害藻华治理方面的发展方向与应用前景,以期为有害藻华的防控提供参考。     

高效溶藻放线茵BS01发酵培养基及发酵条件优化

从漳江口红树林区采集的沉积物样品中分离到1株放线菌菌株BS01 Brevibacterium sp.,其胞外活性产物对塔玛亚历山大藻Alexandrium tamarense具有明显的溶藻作用.采用单因素及均匀设计,通过摇瓶培养对BS01产溶藻活性物质的发酵培养基及发酵条件进行优化.结果表明,在可溶性淀粉为碳源、硝酸钠为氮源、装液量为40%、起始pH值为7.5、培养温度为28℃、转速为150r·min-1、振荡培养时间为48h的条件时,BS01发酵产物的杀藻活性最强.通过均匀设计进行最佳发酵培养基及培养条件优化的结果为:可溶性淀粉为20g·L-1,硝酸钠为0.5g·L-1,pH为7.7,温度为27.2℃.研究结果为杀藻活性物质高效提取及杀藻机制研究奠定了基础.     

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.