椒江口沉积物中细菌多样性初步研究

通过常规分离纯化、鉴定和构建细菌克隆文库的方法,研究椒江口三个站点沉积物中细菌的多样性,并对其进行系统发育分析。可培养细菌的形态学及API鉴定结果显示杀鲑气单胞菌是优势种,典型细菌16S r DNA分子鉴定结果表明γ-变形菌纲和厚壁菌门为主要类群。未培养细菌克隆文库的序列分析结果表明:细菌主要包括变形菌门、酸杆菌门、绿弯菌门、芽单胞菌门、硝化螺旋菌门、CFB群、放线菌门、厚壁菌门等8个类群;其中C0站点克隆子主要属于γ-变形菌纲及绿弯菌门;C1站点克隆子以α-变形菌纲及γ-变形菌纲为主;C2站点克隆子主要属于放线菌门及α-变形菌纲。综合可培养及未培养结果,可发现椒江口沉积物中γ-变形菌纲为典型优势类群,且相当数量的克隆子其且相当数量克隆子的相似序列来自重金属或石油烃污染的沉积环境。     

长江口可培养耐(嗜)盐细菌群落结构研究

针对长江口10个站位样品,采用HM培养基和ZMCA培养基分离培养耐(嗜)盐菌,通过PCR方法获取156株细菌的16S rDNA序列。其中,由HM培养基分离获得136株菌株,可划分为33个分类单元(OUT),分属α-变形菌纲(Alphaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)和厚壁菌门(Firmicutes)的芽孢杆菌纲(Bacilli)3个类群,其中γ-变形菌纲为优势耐(嗜)盐细菌;靠近海岸的站点M5-1,N6-2和M4-1耐(嗜)盐细菌均属于厚壁菌门,而远离海岸的站点耐(嗜)盐细菌均属于变形菌门。以ZMCA培养基分离获得的20株菌株分属α-变形菌纲和放线菌纲(Actinobacteria),可划分为10个OTU。由HM培养基分离获得的菌株中,70.5%属于耐盐菌,29.5%属于中度嗜盐菌;由ZMCA培养基分离获得的菌株均为耐盐菌。HM培养基分离得到的136株细菌的硝酸盐和亚硝酸盐还原性分析表明,亚硝酸盐还原菌数量仅为硝酸盐还原菌的1/3左右。     

南太平洋环流区底层水可培养细菌多样性研究

为研究南太平洋环流区底层海水可培养细菌的多样性,通过IODP 329航次获得了该区域7个站点的底层海水样品,利用传统的分离培养法获得菌株后,进行16SrDNA测序及系统发生分析。结果表明,从南太平洋环流区7个站点的底层水中分离出174株深海细菌,这些菌株属于4个门,30个属,78个种。其中γ-变形菌纲(γ-Proteobacteria)有143株,在数量和种类方面均占主导地位;α-变形菌纲(α-Proteobacteria)7株,β-变形菌纲(β-Proteobacteria)2株,厚壁菌门(Firmicutes)11株,放线菌门(Actinobacteria)6株,拟杆菌门(Bacteroidetes)5株。优势属有假交替单胞菌属(Pseudoalteromonas)、交替单胞菌属(Alteromonas)、弧菌属(Vibrio)、盐单胞菌属(Halomonas)等10个属;优势种有居珊瑚假交替单胞杆菌(Pseudoalteromonas paragorgicola)、子午盐单胞菌(Halomonas meridiana)、坎氏弧菌(Vibrio campbellii)、西班牙交替单胞菌(Alteromonas hispanica)、河豚毒素假交替单胞菌(Pseudoalteromonas tetraodonis)、琼氏不动杆菌(Acinetobacter junii)等12个。在7个站点中,位于环流边缘的U1371站点分离出的菌株数量明显多于其他站点,且是7个站点中唯一1个包含了分离出的所有6个门类的站点,多样性最高;而U1369和U1370站点都只分离出γ-变形菌纲1个门类。此外,9株细菌可能为海洋细菌新属或新种。     

黄海冷水团可培养细菌的多样性研究

本文研究了2008年7月黄海冷水团海域可培养细菌的多样性。AODC和DVC计数总菌数和活菌数分别为(1.3~4.8)×105/mL和(0.6~1.6)×105/mL;2216E平板以菌落计数法可培养细菌浓度为(0.56~2.2)×103cfu/mL。从分离到的475株中选取163株进行16S rDNA扩增,并用HhaⅠ酶进行ARDRA(扩增性rDNA限制酶切片段分析)多态性分析,取114株不同带型测序。结果显示,冷水团细菌归为4个细菌类群:变形菌门(Proteobacteria)、放线菌门(Acti-nobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes),共24个属,其中包括α-,β-,γ-变形菌纲;非冷水团细菌也归为这4个门类,共15个属,但未分离到β-变形菌纲。γ-变形菌纲在冷水团和非冷水团区域的不同深度都是优势菌群,不同的是冷水团区域α-变形菌纲比例较高(26.8%),而非冷水团区域α-变形菌纲比例相对较低(15.6%)。这表明黄海冷水团和非冷水团区域细菌多样性都很丰富,但其群落组成和优势菌群有所不同。此外,16S rDNA测序结果表明,6株细菌可能为海洋细菌新种。     

宁波沿海陆源排污口拟杆菌(Bacteroidetes)分布的特点

采用高通量454焦磷酸测序方法对宁波沿海2个重点排污口、8个一般排污口的20个站位水样进行分析,得到2011年3月、5月、8月、10月份各排污口总体微生物以及拟杆菌纲的分布情况。通过对454测序结果分析,共成功鉴定24个门,主要包括变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和后壁菌门(Firmicutes)等。其中拟杆菌门的变化趋势与梭杆菌门(Fusobacteria)相似,均为随月份的增加而递增的趋势。在拟杆菌门下,共鉴定出3个纲3个目9个科27个属,4个采样月份大部分站点的优势属为黄杆菌纲(Flavobacteria),其次为拟杆菌纲(Bacteroidetes)、鞘脂杆菌纲(Sphingobacteria),通过聚类分析发现,拟杆菌门分布规律主要与排污口类型和季节有一定的关系。通过454测序检测到了极难培养的寡营养细菌鞘脂杆菌,也说明了454测序较传统微生物生态学研究方法的优势,同时针对普雷沃菌属(Prevotella sp.)及黄杆菌属(Flavobacterium sp.)两种具有代表性的属的分布情况进行了分析。     

椒江口化工园区邻近海域沉积物中细菌多样性初步研究

为研究椒江化工园区邻近海域沉积物中细菌结构, 选取四个典型站点, 通过分离纯化、鉴 定和建立克隆文库的方法, 对其进行多样性及系统发育分析。可培养细菌的形态学及API 鉴定结果 显示洋葱假单胞菌及杀鲑气单胞菌杀鲑亚种是优势种, 典型细菌16S rDNA 分子鉴定结果表明厚壁 菌门及γ-变形菌纲为主要类群。非培养细菌克隆文库的序列分析结果表明: 细菌主要包括变形菌门、 酸杆菌门、绿弯菌门、芽单胞菌门、硝化螺旋菌门、CFB 群、放线菌门、厚壁菌门、浮霉菌门等9 个类群; 其中C3 站点克隆子主要属于γ-变形菌纲及δ-变形菌纲; C4 站点克隆子以γ-变形菌纲及酸杆 菌门为主; C5 站点克隆子主要属于δ-变形菌纲及γ-变形菌纲; C6 站点克隆子主要属于γ-变形菌纲及 放线菌门。综合可培养及非培养结果, 可发现椒江口化工园区邻近海域沉积物中γ-变形菌纲为典型 优势类群, 除相当数量的克隆子相似序列来自石油烃污染的沉积环境外, 还有大量克隆子相似序列 来自养殖污染环境。     

山口红树林根际土壤可培养细菌多样性及其活性筛选

为探讨广西山口红树林土壤可培养细菌的多样性，采用纯培养分离技术对采集的25个红树林样品进行可培养细菌多样性分析，挑选117株菌株进行16S rRNA基因测序并构建系统发育树分析。结果表明117株菌株分别属于9个纲20个目27个科35个属，9个纲包括α-变形菌纲（21.37%）、β-变形菌纲（1.71%）、γ-变形菌纲（12.82%）、δ-变形菌纲（0.85%）、噬纤维菌纲（0.85%）、鞘脂杆菌纲（0.85%）、黄杆菌纲（2.56%）、放线菌纲（27.35%）、芽孢杆菌纲（31.62%）。优势属为芽孢杆菌属，占所有菌株的28.20%，此外α-变形菌纲、γ-变形菌纲、黄杆菌纲和放线菌纲中发现了7株潜在的新物种。分离菌株的产酶活性检测表明，山口红树林根际土壤蕴含丰富的产淀粉酶、蛋白酶、葡聚糖、纤维素酶和几丁质酶等生物活性酶的菌株，其中芽孢杆菌纲的产酶菌株数最为丰富。初步研究结果表明广西山口红树林土壤可培养细菌资源丰富，新物种资源多样，是农业微生物开发应用的重要资源库。     

厦门近岸水域海洋附着细菌的群落结构及其动态变化

为研究海洋附着细菌的群落结构及动态变化,在厦门近岸海区进行挂板实验.将无菌玻璃板浸没于海水中,连续放置14 d.分别于放置1 h和7、14 d后取玻璃板上附着生物样品.用细菌通用引物构建16S rRNA基因克隆文库,每个克隆文库随机挑选约40个克隆子测序,序列同源性分析和系统进化分析结果表明,所有的克隆子可分为六大类群:γ-变形菌纲(γ-Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、α-变形菌纲(α-Proteobacteria)、蓝细菌门(Cyanobacteria)和真核硅藻类叶绿体,各类群分别占42.0%、4.5%、2.2%、2.2%、1.1%和45.0%.γ-变形菌纲变形斑沙雷氏菌(Serratia proteamaculans)为优势附着细菌,占测序克隆子的31.5%.这类细菌在1 h样品中的比例超过一半,说明变形斑沙雷氏菌在生物膜形成初期发挥着重要作用.随着挂板时间延长,检测到的细菌类群有所增加:附着7 d后检测到拟杆菌门细菌,附着14 d后检测到厚壁菌门细菌.γ-变形菌纲细菌所占比例随挂板时间的延长而逐渐降低,从挂板1 h的81%降至7 d的21%,14 d的18%.另外,在各阶段的附着样品中,都检测到较多的真核克隆子序列,约占16%～64%.本研究为进一步阐明海洋附着细菌的附着动态及其在生物膜形成过程中的作用奠定了基础.     

莱州湾海域海水和沉积物中的细菌多样性分析

采用16S rDNA文库结合PCR-RFLP分析的手段,对莱州湾水样及沉积物中的细菌多样性和群落特征进行研究。结果表明,莱州湾沉积物细菌类群主要包括变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)两大类群。其中,变形菌门包括α、β、γ3个变形菌纲,所占比例分别为38.7%、3.2%、41.9%。水样中细菌类群主要包括变形菌门(Proteobacteria),酸杆菌门(Acidobacteria),疣微菌门(Verrucomicrobial),浮霉菌门(Plancomycene),拟杆菌门(Bacteroidetes),壁厚菌门(Firmicutes),黏胶球形菌门(Lentisphaeria),绿菌门(Chlorobi)和硝化螺旋菌门(Nitrospira)九大类群。其中,变形菌门包括α、β、δ3个变形菌纲,所占比例高迭60.3%,占有绝对优势。     

长江口及邻近海域表层海水细菌多样性及群落结构

从长江口及邻近海域10 个站点采集表层海水, 通过DAPI 染色计数和克隆文库构建的方法, 对细菌的多样性及群落结构进行初步研究。DAPI 染色计数结果显示长江口及邻近海域表层海水中 细菌丰度总体较高, 生物量变化较大, 从1.16×10⁵(A04 站点)到 1.48×10⁶ cells/mL (B03 站点); 总体 趋势从长江口向外海增加。细菌克隆文库的系统发育分析表明: 细菌序列以变形菌门为主(占总文库 的60.61%), 其中α-变形菌纲是绝对优势类群(50.62%); 其次是拟杆菌门(15.18%)、放线菌门(14.79%) 和蓝细菌门(4.61%), 还有少量厚壁菌门、绿弯菌门、酸杆菌门、浮霉菌门、纤维杆菌门、疣微菌门 等; 另外柔膜菌门和SAR 类群也有发现。文库多样性分析结果显示长江口及邻近海域部分站点表层 海水中细菌多样性显著, 与研究区海域水产养殖活动有关, 并受到区域洋流和海洋化学环境(如低氧 带)的影响。     

Model of the post-Cenozoic evolution of the cryolithozone of the shelf of the western part of the Laptev Sea

The analysis of the structure of the cryolithozone, facies, and thicknesses of the Quaternary sediments and the results of the physicochemical mathematical modeling of the modern shelf of the western part of the Laptev Sea support the influence of the Late Pleistocene glaciations on the heat conditions and the distribution of the permafrost in the area. A ～200-m thick glacier formed under aerial conditions from atmospheric precipitation represented the metamorphosed snow cover. According to the modeling, the long-living (from 60?50 to 10?4 ky) glacier reduced the thickness of the permafrost rocks in the reviewed shelf area for 280–360 m. The Holocene marine transgression additionally decreased the thickness from 50–140 m on the inner shelf to 220–350 m on the outer shelf. The modern submarine cryolithozone 450-0 m thick is wide-spread in the studied region from the coast to the shelf boundary (isobaths of 130–140 m), where it pinches out at a distance of ～380 km from the coast at a depth of ～250 m above the sea level.     

Investigation of the mechanism of the long-term evolution of the carbon cycle in the ecosystem of the Sevastopol bay

We analyze the components of the carbon system of the Sevastopol bay waters and the balance of main sediment-forming substances using the data of field investigations in 1998–2008. The interannual variations of total inorganic carbon and the equilibrium partial pressure of carbon dioxide in the bay water are noted. An increase in the flux of carbon dioxide into the bay and in the content of organic carbon in bottom sediments is revealed, and an explanation of this phenomenon is given. The priority accumulation of organic carbon in the sediments of the bay is established. We assess the interannual variation in the relative abundances of organic and inorganic carbon as an index of the carbon cycle stability.     

Contemporary state of the investigation of the influence of the discharge of rivers on the hydrologic structure of the Black Sea

We generalize and analyze the state of investigations of the influence of river waters on the hydrologic structure of the Black Sea. Specific features of the water, regime and hydrography of rivers flowing into the sea, data on the discharge of rivers, and intrayear and interyear variability of the discharge are presented. We discuss and analyze works studying the transformation of river waters, routes of desalinized sea waters, and their influence on the thermohaline structure and dynamic regime of the sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

双壳贝类血淋巴中儿茶酚胺的检测方法初步研究

研究的目的是探索一种检测双壳贝类血淋巴中儿茶酚胺含量的方法。以栉孔扇贝(Chlamysfarreri)和长牡蛎(Crassostreagigas)为实验对象,研究了取样、抗氧化剂、氧化铝用量以及Tris缓冲液用量等对实验结果的影响。结果表明,高效液相色谱电化学检测器法(HPLC-ECD)可以灵敏高效地对双壳贝类血淋巴中儿茶酚胺进行定性、定量检测。实验表明,取样时间最长不超过1.5min;还原性谷胱甘肽作抗氧化剂效果较好;前处理各试剂与血样量的最适配比为血样量1.5mL,Tris缓冲液(1.5mol/L,pH8.6)1mL和氧化铝25mg;水洗后离心并尽可能吸干氧化铝中的水分。去甲肾上腺素、肾上腺素和多巴胺的回收率分别为53%~69%、47%~73%和48%~56%。     

黄河入海径流对河口海域盐度影响研究

为研究黄河入海径流变化条件下河口附近海域盐度扩散特征,以更好地保护河口海域生物资源多样性,本文以黄河下游利津水文站的长序列实测径流数据资料为基础,利用近海水动力模型FVCOM,分析径流变化对黄河口海域盐度的影响规律。结果表明：黄河口与莱州湾之间存在顺时针的环流系统,在余流作用以及涨落潮方向的影响下,黄河冲淡水长期向莱州湾扩散;丰水期黄河冲淡水几乎影响了整个莱州湾,27盐度锋可以到达莱州湾中部,27等盐线的表层包络面积为2 665.61 km²,占莱州湾的1/4左右,枯水期低盐度水只有向南扩散的趋势,27以下的低盐度水集中分布在黄河口门附近,27等盐线的表层包络面积只有199.65 km²;5月份,随着入海径流量增加,27等盐线扩散的范围、距离、方向都会发生明显变化。在对近海生物资源有迫切保护需求的情景下,适当减少其他用水户供水量以增加入海生态径流量,可以有效改善黄河口海域附近的盐度情况,为生物资源的生长繁殖创造良好条件。     

Modelling of the ecosystem of the Black-Sea shelf zone near the estuary of the Danube

We analyze the results of numerical calculations performed according to the three-dimensional interdisciplinary model of an ecological system of the Black-Sea shelf zone near the estuary of the Danube. The complete system of equations of hydrothermodynamics is solved together with transport equations of the advection-diffusion-reaction type used to describe the transformation of a substance (nitrogen) between the components of the characteristic vectors of the ecosystem: plankton, detritus, and biogenic elements (nitrates). We describe the distinctive features of the circumcontinental distribution of components obtained as a result of numerical experiments and present arguments for the conclusion that the ecosystem of the Danube estuary water area plays the role of a buffer zone between the press of the Danubian biogenic pollutions and the neighbouring areas of the shelf zone and open sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

The influence of the bottom contour on the deformed state of the ice cover due to the motion of the submarine

The authors have previously determined that the effectiveness and failure pattern of the ice cover caused by flexural-gravity waves generated by a submerged body motion near the bottom ice can greatly depend on the depth of the water area. In its turn, the presence of a ledge on the ice surface may affect a wave propagation pattern. This paper presents an experimental study of the bottom contour influence on the deflection and length of flexural-gravity waves. The authors describe a numerical model for the analysis of the deformed state of ice caused by hydrodynamic loads due to a submarine motion, taking into account the bottom contour. The experiments are carried out in the ice tank. The results of calculations and experiments are compared.     

Dynamics of the ice cover and peculiarities of the ice transportation of the sediments at the tidal flats of the Kandalaksha Gulf of the White Sea

Ten years of semistationary studies of the interaction of the ice cover and the coastal relief of the Kandalakhsa Gulf in the White Sea allowed us to reveal the peculiarities of these processes in the warm and cold years and the mechanisms of the origination of ice forms, including ice tents, which govern the character of the tidal flats. Three dynamic fast ice zones are distinguished by their geomorphological location, morphology, and ice dynamics. The dependence of the mechanisms and volumes of the enrichment of the fast ice in the sediments on the coastal zone??s relief was established. At the coast of the Kindo Peninsula (Velikaya Salma Strait), the ice mostly contains fine-sandy sediments. Coarser sediments, including boulders, are occasional and frozen into the ice at the contact of the fast ice and the bottom. At the expense of the ice??s spreading by 4?C6 times in the warm years and 2?C4 times in the cold years, several thousand tons of sediments mostly from the tidal flat??s surface (100?C700 m³ from 1 km of coast in the wide tidal flats and <50 m³ on the steep coasts) are delivered to the water area.     

Relationship between the path of the Kuroshio in the south of Japan and the path of the Kuroshio Extension in the east

A relationship between paths of the Kuroshio and Kuroshio Extension (KE) is investigated, using the satellite-derived altimetry dataset of 1993–2008. When the Kuroshio takes the nearshore nonlarge meander path or typical large meander path and resultantly goes through the deeper channel (about 2500 m) of the Izu-Ogasawara Ridge, the KE path adopts a relatively stable state with the two quasi-stationary meanders. On the other hand, when the Kuroshio takes the offshore nonlarge meander path and then passes over the shallower part of the Ridge (about 1000 m), the KE path tends to be convoluted, i.e., an unstable state.     

A model of the dynamics of the ecosystem of the Sea of Azov

We use many-year archival observations of hydrologic and hydrobiologic state of the Sea of Azov for the identification of the structure of a model of its ecosystem. The ecosystem model, supplemented with expert estimates of bioresource consumption (fish reserve), contamination level, and possible ecologic fines for violation of the sea natural state, is formalized by the method of system dynamics. Hereat, the major part of influence functions is found in terms of observational data with application of self-organization algorithms. We also present the results of simulated experiments with the model of the ecosystem, which enable us to analyse scenarios of its behaviour under the influence of various external factors (wind, river discharge, water exchange with the Black Sea, etc.). Translated by Peter V. Malyshev and Dmitry V. Malyshev