深海产低温碱性淀粉酶菌Halomonas sp.W7的筛选及发酵条件研究

从27份深海沉积物中筛选到30株产淀粉酶细菌,并对其中的W7菌株的产酶条件及酶学性质进行了研究。对W7菌株进行16SrDNA序列分析表明该菌株属于盐单胞菌属(Halomonas)。其最适生长温度为25℃,能适应pH值7～13和盐度0～100的环境条件。该菌株可利用多种碳源,但只在淀粉存在的条件下产酶,可利用有机氮源和无机氮源,但有机氮源更能促进淀粉酶的产生。产酶的最适条件为:25℃,pH10,接种量2%,盐度50,150r/min摇床培养36～48h。粗酶液的最适作用温度为40℃,最适pH值为10。该菌具有反硝化和氨化活性。     

纳米ZnO对1株异养硝化-好氧反硝化菌Halomonas sp. KGL1的生物胁迫效应研究

本文通过纳米ZnO(ZnO-NPs)对具有高效脱氮能力的异养硝化-好氧反硝化菌Halomonas sp. KGL1的短期暴露实验,探讨在不同作用浓度下(0,1,10,50 mg·L~(-1)) ZnO-NPs对菌株的生物胁迫效应。结果表明,ZnO-NPs破坏菌株Halomonas sp. KGL1的细胞膜完整性并改变其粘滞性,使菌株形态结构改变,菌体发生团聚;同时诱导该菌株细胞产生活性氧(Reactive Oxygen Species,ROS),对菌株细胞产生氧化胁迫,进一步损伤菌株细胞,抑制菌株的生长和脱氮能力,且ZnO-NPs浓度越高,该菌株受胁迫程度越强。不同浓度的ZnO-NPs对菌株Halomonas sp. KGL1的NH~+_4-N去除率无显著影响,而其NO~-_3-N、NO~-_2-N的去除效率显著降低。研究结果可为提高海水养殖废水等高盐含氮废水中脱氮菌株的抗ZnO-NPs胁迫能力的理论研究和实际应用提供科学依据。     

产酯酶海洋中度嗜盐菌Halomonas sp.LYG1-1的分离鉴定及其酶学性质

以从海南三亚近海海水中采集的样品为研究对象,利用平板筛选法筛到一株产酯酶的中度嗜盐菌菌株LYG1-1。通过形态学、生理生化及16S rRNA序列分析,发现该菌株为革兰氏阴性菌,其最适生长温度为30℃,最适生长pH为7.5,最适NaCl浓度为10%。16S rRNA基因序列分析显示,菌株LYG1-1与Halomonas salina亲缘性最近,16S rRNA基因序列相似性为99.72%。在含10%NaCl的0.05mol·L-1Tris-HCl(pH 8.0)缓冲液中,45℃条件下作用,酶活性最高,40℃处理24h仍保持83%的活性。金属离子浓度为5mmol·L-1时,Ca2+、Mg2+对菌株LYG1-1酶活具有明显的激活作用,Hg2+表现出强烈的抑制作用。     

中国对虾体内1株益生菌的筛选与初步鉴定

为给对虾专用益生菌制剂的研制与应用提供理论依据和基础数据,研究了从白斑病毒(WSSV) 耐过中国对虾(Fenneropenaeus chinensis)肠道中分离的1株海洋细菌B12的益生特性和安全性,并结合形态观察、生理生化特征和16S rDNA序列分析对菌株B12进行了分类鉴定.结果表明:菌株B12能抑制对虾致病性哈维氏弧菌 (Vibrio harveyi)和副溶血弧菌(V. parahaemolyticus)的生长,不分泌溶血素.对β-内酰胺类、头孢类、氨基糖苷类、喹诺酮类抗生素均敏感;对大环内酯类、四环素类抗生素均耐药;对糖肽类抗生素中度敏感.毒性试验表明菌株B12对中国对虾幼体没有明显毒副作用.该菌为革兰氏阴性可动杆菌,鞭毛极生,菌体大小为(0.5～0.6)×(1.1～1.2) μm,接触酶、氧化酶阳性,葡萄糖发酵产酸,能还原硝酸盐,产淀粉酶,不产明胶酶,不能利用丙二酸.16S rDNA的部分序列分析显示菌株B12与嗜盐单胞菌(Halomonas sp.)SB J85具有98.15%的相似性.形态观察、生理生化特征和16S rDNA序列分析证实B12为嗜盐单胞菌属(Halomonas sp.).     

一株海洋聚磷菌YSR-3的分离与鉴定

采用平板分离法,从黄海海域分离到一株聚磷菌,编号为YSR-3,对其进行了部分生理生化特性以及分子特性的研究。结果表明,YSR-3菌体呈杆状,大小为3·5μm×1μm,革兰氏染色阴性,好氧生长;显微镜下观察能运动。最适培养条件为:氯化钠浓度3%,生长温度为24℃,起始pH值为6·5。该菌能在细胞内累积多磷酸盐形成异染粒。16SrDNA鉴定结果表明,YSR-3属于γ-变形菌纲、盐单胞菌属(Halomonas)。与盐单胞菌属中的其他已知种比较,YSR-3具有其特性:对氯霉素和卡那霉素敏感、淀粉水解呈阳性、反硝化和几丁质降解呈阴性、能将葡萄糖作为唯一碳源和能源。     

盐单胞菌属一新种的分离与鉴定

对即墨市大桥盐场的底泥样本进行微生物的可培养研究,分离出1株革兰氏阴性的中度嗜盐菌YC-SA28T.16SrRNA序列比对及系统发育分析结果表明.YCSA28T可能为γ-变形菌纲盐单胞菌属的新种,通过系统分类学研究发现,YCSA28T的16SrRNA序列与亲缘关系最近的盐单胞菌属模式菌株Halomonas ventos...     

浒苔(Enteromorpha prolifera)生长及孢子释放的生态因子研究

通过考察浒苔属浒苔藻体生长及孢子释放的生态因子条件,确定其各单因子的适应范围及最适范围.结果表明:浒苔藻体生长对盐度、温度、光照强度和酸碱度的适应范围分别为 16～40、10 ℃～30 ℃、＞9 μmol/m2s、6～10,最适范围分别为 24～28、20 ℃～25 ℃、＞18 μmol/m2s、8～9,在盐度、温度、光照强度和酸碱度分别为24、25 ℃、72 μmol/m2s、8 的条件下达到最大生长量;浒苔释放孢子对盐度、温度、光照强度和酸碱度的适应范围分别为 12～40、15 ℃～35 ℃、＞9 μmol/m2s、6～10,最适范围分别为 28～40、20 ℃～35 ℃、＞36 μmol/m2s、8～9,在盐度、温度、光照强度和酸碱度分别为32、35 ℃、144 μmol/m2s、9 的条件下达到最大释放量.浒苔藻体生长及孢子释放的各单因子适应范围基本相同,最适范围区别明显.     

产碱性纤维素酶海洋细菌的筛选、鉴定及酶学性质研究

用CMC平板筛选方法,从青岛近海海域海水中分离出一株产碱性纤维素酶的海洋菌株QM11,经16S rDNA鉴定,该菌株为Cytophaga fucicola.对该菌的生物学特性研究表明,其最适生长温度为27℃,生长温度范围为4～48℃,为耐冷菌;在pH7.0～8.0、含3.0%NaC1的培养基条件下,最适宜菌株生长和产酶;QM11所产碱性纤维素酶最适反应温度为40℃,最适反应pH为9.0,在碱性条件下具有较高的酶活性和较好的稳定性.Mn2+、Fe3+对酶反应具有促进作用,Cu2+、Pb2+对酶反应具有抑制作用.     

深海来源芳烃类降解菌筛选与盐单胞菌芘降解能力测定

海洋来源的菌株在高盐污水类修复处理中有重要的应用前景,而烃类物质是污水中的重要组成部分,其中含有多个苯环的芳香烃族化合物,对人类和环境具有较大的毒性,而目前关于多环芳烃(PAHs)厌氧代谢的相关报道相对较少。本研究以PAHs(萘、菲和芘等)为唯一碳源和能源,通过厌氧富集对太平洋深海沉积物中的PAHs降解菌展开研究。富集物细菌菌群分析结果表明,盐单胞菌属(Halomonas)、海旋菌属(Thalassospira)、海杆菌属(Marinobacter)、海洋杆菌属(Oceanobacter)和食烷菌属(Alcanivorax)等是主要的功能类群;其中盐单胞菌属是最主要的功能类群。通过筛选分离获得一高效烃降解盐单胞菌株,鉴定并命名为泰坦尼克盐单胞菌(Halomonas titanicae) PA16-9,该菌与模式菌株Halomonas titanicae BH1^T的16S rRNA相似性为99.52%。基于16S rRNA基因序列比对搜寻NCBI数据库,发现Halomonas titanicae广泛存在于废水、活性污泥、油田、湿地等有机质复杂区域,生态位分布极广,可...     

一株新型异养硝化-好氧反硝化菌GJWA3的脱氮性能及定量检测

从水产养殖水体中分离出一株具有异养硝化-好氧反硝化能力的菌株Halomonas sp.GJWA3,该菌株在单一氮源条件下(10 mg·L-1)对NH+4-N、NO-2-N和NO-3-N的48 h去除率分别为96.44％、99.42％和78.27％,氮平衡分析结果表明该菌株能够去除水体中大部分无机氮.该菌株在pH为7.0...     

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.     

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.     

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.     

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.     

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.