潮间带沉积物厌氧烃降解细菌的多样性及Desulfovibrio subterraneus ND17的分离鉴定

潮间带作为海陆交界处,易受到来自海洋的石油污染,且各类石油烃进入沉积物后的降解过程尚不清楚。前人在各类生境中对好氧微生物烃降解方面已有较多研究,但对近海潮间带环境中的厌氧烃降解鲜有报道。本研究对青岛女岛湾潮间带沉积物深层样品以混合烃（中长链烷烃、多环芳烃）为碳源,硫酸盐作为电子受体进行厌氧富集培养。富集菌群的细菌多样性表明在混合烃作为碳源的作用下,优势菌群转变为脱硫叠球菌科（Desulfosarcinaceae）、脱硫杆菌科（Desulfobacteraceae）等具有石油烃降解潜力的硫酸盐还原菌。经分离纯化得到一株厌氧烃降解菌ND17,与地下脱硫弧菌属模式种Desulfovibrio subterraneus HN2T 16S rRNA基因序列的相似度为99.93%。进一步实验表明,菌株ND17在厌氧条件下对二十四烷和菲的降解率可分别达到53.9%和35.7%。这也是首次对脱硫弧菌属单菌在厌氧条件下进行石油烃降解的研究。脱硫弧菌作为一种广泛分布在厌氧环境的细菌,本研究为进一步认识其在海洋石油污染环境中的修复潜力提供了支撑。     

Relationship between hydrocarbons and bacterial activity in mediterranean sediments: Part 2—hydrocarbon degrading activity of bacteria from sediments

In marine sediments from the Mediterranean coast polluted by oil spills (Etang de Berre) high bacterial activity is responsible for self-purification, samples collected from these areas showing a bacterial density several orders of magnitude higher than samples from non-polluted areas (Isle des Embiez). Between 60 and 80 % of the heterotrophic bacteria in the polluted areas are hydrocarbon-degrading, compared with 0.01 % in the non-polluted area.Several strains of hydrocarbon utilisers were isolated, a mixture of strains from each biotope was taken to represent a ‘biotope population’ and their activity towards different types of hydrocarbons (n-alkanes, iso-alkanes, cycloalkanes, aromatics) was determined.With ammonia as the nitrogen source the ‘biotope population’ from the polluted area degraded hydrocarbons much more readily than the ‘biotope population’ from the non-polluted area. With nitrate as the nitrogen source the degradation was much reduced, and, for some hydrocarbons, ceased in both ‘populations’.Individual hydrocarbons encouraged the growth of individual strains in the ‘biotope population’. In natural mixtures of hydrocarbons all strains grew well and degraded aliphatics. There was a good correlation between the respiratory activity of the ‘biotope population’ and the ability of the ‘population’ to utilise hydrocarbons for growth.     

南极微生物对低温生活污水处理效果的初步研究

为了提高低温废水处理效果,筛选：和鉴定具有显著低温生活污水降解作用的南极微生物,并对其低温生活污水的处理效果进行了初步研究。实验结果表明,在0～5℃时。南极低温酋群对有机物的降解率显著高于中温菌的去除率,南极耐冷菌对NH3-N及TP均有一定的去除率。从南极低温茼群中筛选到5株蛋白质降解功能菌,4株脂肪降解功能菌,1株淀粉降解功能菌,2株烷烃降解功能菌和1株芳香烃降解菌,并通过16SrDNA序列对它们进行了鉴定,明确其菌属。这为寻求解决低温环境中废水处理的有效方法奠定了基础。     

南极菲尔德斯半岛可培养土壤微生物多样性及理化性质鉴定

为了探索南极可培养土壤微生物的多样性,本研究对中国第31次南极科学考察采集自南极菲尔德斯半岛的5份土壤样品进行了细菌、真菌的分离培养。选择形态差异较大的细菌和真菌进行了16S rDNA和ITS鉴定及系统发育分析。最终共鉴定得到20个属的33株细菌和6个属的8株真菌,其中8株细菌,10株真菌序列相似性较低,可能是新种。该结果表明南极菲尔德斯半岛地区具有丰富的微生物多样性,其中假单胞菌属的细菌居多。对分离得到的细菌和真菌分别进行理化性质和胞外酶活性鉴定,实验结果显示,分离获得的细菌和真菌绝大多数可产生水解酶类并同化多种碳源。初步认定这些微生物在参与南极物质代谢、适应南极极端环境方面发挥作用。本研究丰富了对南极菲尔德斯半岛可培养土壤微生物多样性的认识,并筛选获得了一些产低温酶特性的菌株,这为极地微生物资源的利用研究奠定了基础。     

Experimental degradation of crude oil by marine bacteria

The ability of biodegradation for crude oil was examined for ten strains of marine bacteria. With regard to their degradation ability forn-alkanes the microorganisms tested could be divided into four groups, although the same result was not observed for microbial degradation of aromatic hydrocarbons obtained by the UV measurements.     

南海沉积物细菌胞外蛋白酶在家族水平上的多样性

海洋产蛋白酶细菌及其分泌的胞外蛋白酶在降解有机氮以推动海洋氮循环进行方面发挥着重要作用,但目前对这二者多样性的认识非常有限。本研究自南海沉积物中筛选得到90株产蛋白酶细菌,并通过N-端氨基酸序列,分析了其胞外蛋白酶在家族水平上的多样性。16S rRNA基因序列分析的结果表明,筛选的产蛋白酶细菌均属于Gammaproteobacteria纲,且大多数属于Alteromonadales目和Vibrionales目的不同属。对其中14株菌株的14个胞外蛋白酶的N-端氨基酸序列分析表明,所有这些蛋白酶属于金属蛋白酶的M4家族或丝氨酸蛋白酶的S8家族。本研究提供了海洋沉积物产蛋白酶细菌在类群及其胞外蛋白酶在类型上的新细节,这将有助于全面了解微生物酶促降解海洋沉积物有机氮的过程和机理。     

Degradation of lubricating oils by marine bacteria observed by quantitative mass spectrometry

Bacterial degradation of the hydrocarbons of lubricating oils was investigated by mass spectrometric analysis which gives both total amount and the composition of hydrocarbon types of residual oil. An unused lubricating oil, which mainly consisted of hydrocarbon types with only a small percentage ofn-alkanes, was degraded by marineBacillus sp. andPseudomonas sp. with 55 % and 25 % decreases in 10 days, respectively. Susceptibility of respective hydrocarbon types to biodegradation was in the following order: alkanes > non-condensed cycloalkanes, mono-aromatics > condensed cycloalkanes. A used lubricating oil of different brand showed a larger decrease than the unused oil. Both species of bacteria degraded large portions of alkane fraction of Arabian light crude oil.     

海洋附着细菌对冠瘤海鞘幼体附着和变态的影响

从冠瘤海鞘、皱瘤海鞘及乳突皮海鞘的被囊表面及其附着基和附近的海水中分离到290株细菌,根据其形态和生理生化特征,将其鉴定至弧菌属、肠杆菌属和无色杆菌属等10个属.分析海鞘被囊表面及其附着基和海水中的细菌组成,并从各属中挑出的29株细菌中分离筛选出附着能力较强的9株.将这9株细菌分别制成不同的单种细菌黏膜及9种细菌混合黏膜,检测细菌黏膜对冠瘤海鞘幼体附着和变态的影响.结果表明不同细菌黏膜对冠瘤海鞘幼体的附着和变态起不同的作用.无色杆菌属H-13显著地促进海鞘的附着和变态;弧菌属的H-4对海鞘的附着和变态均起抑制作用;弧菌属H-9明显地促进附着却强烈抑制海鞘幼体变态;其余细菌仅仅对海鞘幼体附着或变态的一个过程起作用或是对两者都没有明显效果.     

Evidence for the decoupling of dissolved, particulate and surface microlayer hydrocarbons in Northwestern Atlantic continental shelf waters

Detailed sampling of the water column and subsequent high-resolution gas chromatographic analyses of surface film, particulate and dissolved hydrocarbons from the Georges Bank region off the New England coast have revealed large spatial and temporal heterogeneity in the source of hydrocarbons. Dissolved hydrocarbons, largely of a petroleum-related origin, surface film hydrocarbon also of a petroleum-related origin but different from the dissolved fraction, and bulk-water particulate matter of a mixed biogenic and petrogenic origin are all decoupled with respect to source and quantity in the region.Surface film (microlayer) hydrocarbons are found in greater concentrations than the dissolved fraction, which in turn is greater than the particulate hydrocarbon quantities in the water column. Although the microlayer appears enriched in hydrocarbons relative to the bulk water, the nature of the hydrocarbons is quite different, indicating that the use of a microlayer enrichment factor may not be appropriate in these waters.     

金顶铅锌矿床油气地球化学特征及其成矿作用探讨

金顶铅锌矿区上三叠统三合洞组角砾状灰岩为其主要容矿岩层 ,最近在与金顶矿床共生的灰岩角砾岩、深灰色泥晶灰岩和灰色云化灰岩中发现多层稠油储层及油气包裹体。包裹体烃类地球化学研究表明存在多源性母质输入 ,油气母源类型以藻类及细菌等微生物贡献为主 ,沉积水体介质为高盐度、较强还原环境 ,源岩演化处于低成熟～成熟。铅锌金属与稠油的共生、金属硫化物的有机生物硫特征、有机质富集地区矿化强烈等均表明有机质与金属成矿存在相互作用。按照不同世代有机质与金属成矿作用机理 ,本区有机成矿以配合作用及吸附、还原作用为主 ,有机质的存在增加了铅锌离子的溶解度 ,促进了铅锌金属离子的迁移和富集 ,有利于产生铅锌金属沉淀的还原环境。     

The effects of oil exploration and production in the northern North Sea: Part 2—microbial biodegradation of hydrocarbons in water and sediments, 1978–1981

The potential of heterotrophs to degrade aromatic hydrocarbons derived from oil in water and sediments in the northern North Sea around the Brent, Beryl, Forties and Murchison oilfields and at a number of stations, from the Forties field to the Firth of Forth, was estimated using 1-¹⁴C naphthalene and 7, 10-¹⁴C benzo(a)pyrene. The degradation of uniformly labelled ¹⁴C mixed 1-amino acids was used as a measure of total heterotrophic activity. Results showed that microorganisms have the potential to degrade the smaller aromatic hydrocarbon molecules rapidly in the water column and in surface sediments. Close to the Beryl platform, where diesel-washed drill cuttings have been continuously dumped, the rate of input of hydrocarbons to the sediment has exceeded the rate of degradation. Mineralization of benzo(a)pyrene, estimated in the sediments only, was minimal. Close to production platforms the biodegradation rate of mixed 1-amino acids showed no increase comparable with that found for naphthalene, suggesting that existing microbial populations had not increased but adapted to degrade oil in water and sediments.     

厦门海区潮间带海洋动植物共附生微生物的抗菌活性

从厦门海区潮间带石纯、浒苔、江蓠、海兔、海葵及鲨鱼肠道分离得２８９株共附生微生物,以８种敏感菌为指示菌,从中获得拮抗活性的幼苗２８株,放线菌１７株,真菌８株。拮抗细菌以假单胞菌（３９％）、弧菌（２５％）和芽直菌（１４％）为主,主要为抗革兰氏阳性细菌,抗真菌的菌株比例较低。拮抗放线菌分属于链霉菌和小单孢菌,其抗革兰氏阳性菌、阴性菌和真菌的菌株比例分别为６５％、４７％和２３％。拮抗真菌分青霉属和芽枝霉     

浙江镇海潮间带沉积物中可培养微生物初步研究

针对浙江镇海潮间带沉积物样 品,采用纯培养法分离培养海洋微生物,并基于16S rRNA基因序列,开展系统发育学研究,分析沉积物细菌群落结构及多样性。分离获得细菌39株,16S rRNA基因序列分析表明,这些菌株分别属于厚壁菌门Firmicutes(51. 3%)、变形杆菌门Proteobacteria(30.8%)、拟杆菌门Bacteroidetes(15. 4%)和放线菌门Actinobacteria(2. 6%)4个类群。厚壁菌门和变形杆菌门的菌株主要归属于芽孢杆菌纲Bacilli和a-变形杆菌纲Alphaproteobacteria,它们可能在近海海洋生态系统的元素地球化学循环中发挥了重要作用。此外,部分菌株16S rRNA基因序列与已报道物种的相似性较低,可能代表了新的分类单元。研究丰富了浙江近海微生物种质资源,并拓展了对浙江近海可培养细菌多样性的认识,为今后开发利用海洋微生物资源积累了资料。     

南极罗斯海区域可培养微生物分离鉴定及产低温酶能力初步筛选

为了利用南极微生物资源、探索南极罗斯海区域可培养微生物多样性,利用传统平板培养法对中国第33次南极科学考察采自南极罗斯海6个站位的海洋沉积物样品进行了细菌、真菌的分离培养。经细菌16S rRNA、真菌ITS基因序列检测及系统发育分析,共获得5个属的36株细菌和6个属的29株真菌。其中嗜冷杆菌属为优势细菌类群,枝孢属为优势真菌类群。该结果表明南极罗斯海区域具有丰富的微生物多样性。细菌API 20 NE生理生化及真菌产胞外酶活性实验显示,分离得到的细菌和真菌绝大多数都有低温酶活性。研究结果为南极罗斯海区域可培养微生物的多样性认识和低温酶产生菌资源获取与开发利用提供了支撑。     

南极石油烃降解嗜冷菌的筛选及其降解特性的研究

从385株南极海洋细菌中筛选出2株石油烃降解菌,并对其降解特性进行了初步研究。以柴油为唯一碳源进行降解实验的结果表明,南极嗜冷菌NJ276和NJ341在5℃、20d内对柴油的降解率分别达到23．47％和32．15％,在15℃、20d内降解率分别达到43．95％和62．47％,其降解能力随着培养温度的升高而显著增强;石油烃降解残油组分的GC～MS分析表明,柴油经过NJ276降解后的残油组分中能检测到C15～C21七种烷烃,柴油经过NJ341降解后的残油组分只能检测到少量C16,C17和C18三种烷烃。对它们进行16S rDNA基因序列的同源性和系统发育分析表明,菌株NJ276属于假交替单胞菌属（Pseudoalteromonas）,NJ341属于科尔韦尔氏属（Colwellia）。     

A study of crude oil‐degrading bacteria from mangrove forests in the Persian Gulf

Mangroves are coastal ecosystems, found in tropical and subtropical regions around the world. They are found in the transitional zones between land, sea, and rivers. Petroleum hydrocarbons are the most common environmental pollutants, and oil spills pose a great hazard to mangroves forests. This research was focused on the isolation and characterization of crude oil‐degrading bacteria from mangrove ecosystems at the Persian Gulf. Sixty‐one crude oil‐degrading bacteria were isolated from mangrove samples (plant, sediment, and seawater) that enriched in ONR7a medium with crude oil as only carbon source. Some screening tests such as growth at high concentration of crude oil, bioemulsifier production, and surface hydrophobicity were done to select the most efficient strains for crude oil degradation. Molecular identification of strains was carried out by amplification of the 16S rRNA gene by PCR. The results of this study were indicated that the quantity of crude oil‐degrading bacteria was higher in the root of mangrove plants compare to other mangrove samples (sediment and seawater). Also, identification results confirmed that these isolated strains belong to Vibrio sp. strain NW4, Idiomarina sp. strain BW32, Kangiella sp. strain DP40, Marinobacter sp. strain DW44, Halomonas sp. strain BS53, and Vibrio sp. strain DS35. The application of bioremediation strategies with these bacteria can reduce crude oil pollution in this important marine environment.     

Community diversity of bacteria in digestive tract of mud snail (Bullacta exarata Philippi) and its rearing shoal

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Bacterial communities fluctuate in abundance and diversity under simulated oil-contaminated seawater conditions

Marine bacteria have recently been identified as a potent solution for petroleum hydrocarbon degradation in response to hazardous oceanic oil spills. In this study, a mesocosm experiment simulating a petroleum spill event was performed to investigate changes in the abundance, structure, and productivity of bacterial communities in response to oil pollution. Cultured heterotrophic bacteria and total bacteria showed a consistent trend involving an immediate decrease in abundance, followed by a slight increase, and a steady low-level thereafter. However, the changing trend of bacterial productivity based on bacterial biomass and bacterial volume showed the opposite trend. In addition, the density of oil-degrading bacteria increased initially, then subsequently declined. The change in the bacterial community structure at day 0 and day 28 were also analyzed by amplified ribosomal DNA restriction analysis (ARDRA), which indicated that the species diversity of the bacterial community changed greatly after oil pollution. Alphaproteobacteria (40.98%) replaced Epsilonproteobacteria (51.10%) as the most abundant class, and Gammaproteobacteria (38.80%) became the second most dominant class in the whole bacterial community. The bacterial communities in oil-contaminated seawater (32 genera) became much more complex than those found in the natural seawater sample (16 genera). The proportion of petroleum-degrading bacteria in the oil-contaminated seawater also increased. In this study, culture-dependent and culture-independent approaches were combined to elucidate changes in both bacterial productivity and community structure. These findings will contribute to a better understanding of the role that bacteria play in material cycling and degradation in response to oil pollution.     

三亚湾珊瑚来源虫黄藻不同株系微环境中微生物群落结构的差异比较分析

虫黄藻、细菌和造礁石珊瑚有着密切的共生关系, 但虫黄藻藻际细菌群落尚未得到广泛研究。本研究对5个属的6株虫黄藻进行了离体培养, 其中2株为悬浮型虫黄藻(E型), 4株为贴壁型虫黄藻(A—D型)。通过采集藻株培养物3种粒径的样本开展细菌群落分析, 分别为0.2~3μm(自由生活)、>3μm(附着于藻体或颗粒物)与Settling(沉底贴壁藻体上)。结果发现, 2种生活方式的藻株藻际细菌群落具有显著差别, 贴壁型藻株细菌群落的物种丰富度显著高于悬浮型藻株。发现7个属的细菌广泛存在于所有的藻株中, 它们代表了A—E型虫黄藻藻际细菌的核心类群。对3种粒径样本的核心细菌群落比较发现, 自由生和颗粒附着生的核心细菌群落十分相似, 但均与沉底贴壁样本差异明显。     

我国近海中度嗜盐菌的分离筛选及其产酶多样性分析

为了解我国近海中度嗜盐菌系统发育多样性及产酶特性,从天津、山东、江苏、福建、海南等地近海采集样品为研究对象,分离筛选得到108株中度嗜盐菌,其中有26株至少产一种酶。通过对其形态特征、生理生化、16SrRNA基因序列及系统进化树进行分析,将26株菌鉴定为细菌域的Halomonas、Idiomarina、Virgibacillus、Pontibacillus、Oceanobacillus、Halobacillus和Marinilactibacillus属的菌株。它们与相应属的模式菌株16SrRNA基因序列相似性在97%~100%不等,其中有些菌株可能代表不同的分类单元。底物特异性试验表明,分离的26株中度嗜盐菌13株产蛋白酶,19株产淀粉酶,13株产酯酶,4株产纤维素酶。其中6株产3种酶,11株产2种酶。研究结果表明中度嗜盐菌具有系统发育和产酶多样性,同时蕴藏着较多新的微生物类群,为嗜盐微生物资源应用开发提供了参考。