海洋环境中的厌氧氨氧化细菌与厌氧氨氧化作用

厌氧氨氧化是细菌在厌氧条件下将氨氮氧化成氮气的过程,主要应用在污水处理反应器中,最近几年发现在海洋环境中也广泛存在,并在海洋氮循环过程中发挥了重要作用,代表了海洋中一个巨大的氮汇,对碳循环和全球气候变化也有重要影响.梯烷膜脂结构独特,是厌氧氨氧化细菌的化学生物标志物,具有化学分类与古海洋学应用潜力.厌氧氨氧化作用及厌氧...     

海洋环境中的厌氧氨氧化细菌与厌氧氨氧化作用

厌氧氨氧化是细菌在厌氧条件下将氨氮氧化成氮气的过程,主要应用在污水处理反应器中,最近几年发现在海洋环境中也广泛存在,并在海洋氮循环过程中发挥了重要作用,代表了海洋中一个巨大的氮汇,对碳循环和全球气候变化也有重要影响。梯烷膜脂结构独特,是厌氧氨氧化细菌的化学生物标志物,具有化学分类与古海洋学应用潜力。厌氧氨氧化作用及厌氧氨氧化细菌已成为海洋生物地球化学、微生物学、有机地球化学等研究领域的热点。     

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.     

Effects of fixation and storage on flow cytometric analysis of marine bacteria

Flow cytometry (FCM) is now becoming a routine tool for the enumeration and optical characterization of bacteria in marine environments. We investigated the effects of sample fixation and storage upon flow cytometric determination of marine bacteria. Fixed and unfixed seawater samples were analyzed by FCM immediately aboard ship and/or later in the laboratory, and the appearances of the fluorescence signals and bacterial counts of these samples were compared. Fixation and storage led to the formation of multiple peaks in fluorescence histograms; this was also seen in 22 out of 36 samples frozen in liquid nitrogen. Fixation did not, but storage did induce a decrease of bacterial counts: a rapid decrease during the first 3 days followed by a slower decline. The decline of cell numbers in stored samples was expressed by a regression model. Our studies indicate that precaution is necessary when interpreting the data from fixed and/or stored marine bacterial samples analyzed by FCM. The possibility that the procedure of fixation and storage leads to the appearance of high DNA and low DNA bacterial groups should be considered.     

海洋微微型蓝藻与异养细菌相互作用的研究进展

菌藻相互作用是海洋生态学领域研究的重要方向之一.海洋微微型蓝藻(Marine picocyanobacteria)是遍布全球海洋的重要初级生产者,在全球碳循环和微食物网中发挥重要作用.原绿球藻属(Prochlorococcus)和聚球藻属(Synechococcus)是海洋微微型蓝藻最重要的两个类群.原位调查和培养实验...     

从养殖贝类和大菱鲆分离的海洋发光弧菌的分类地位分析

海洋发光弧菌是海水养殖中重要的条件致病菌。从福建省晋江市附近水产养殖场花蛤、牡蛎软组织及波罗地海大菱鲆肠道内含物中分离得到16 株发光细菌, 采用ARDRA（amplified ribosomal DNArestriction analysis）、16S rDNA 序列测定、Biolog 碳源代谢分析及药敏试验等方法对...     

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.     

海洋源拮抗细菌的分离、筛选及1株拮抗芽孢杆菌的鉴定

对来自于不同海域的红树树皮、海苔(海洋生物样)和海泥(海洋底质)样品进行细菌分离,并采用管碟法测试了分离出的海洋细菌对金黄色葡萄球菌Staphylococcus aureus、枯草芽孢杆菌Bacillus subtilis、大肠杆菌Escherichia coli、白色念珠菌Candida albicans、黑曲霉As...     

Phylogenetic analysis and biological characteristic tests of marine bacteria isolated from Southern Ocean (Indian sector) water

Fifty-seven bacteria were isolated from Southern Ocean(Indian sector) water samples which were collected from different latitude and longitude of the ocean. All the isolates were able to grow at 4°C, 20°C, 37°C and tolerable Na Cl concentration up to 13.5%(w/v). 29 out of 57 isolates were identified using 16 S rDNA amplification and the sequences were submitted to National Center for Biotechnology Information(NCBI). All the isolates were classified by using Ribosomal Database Project(RDP) and found that isolates belongs to Proteobacteria and Bacteriodes. The average G+C content was 56.4%. The isolates were screened for the presence of extracellular enzymes, viz. amylase, catalase, urease, esterase, lipase and protease. The disc diffusion method is used to screen antibiotic production by the isolates against four pathogenic bacteria, viz. Salmonella typhimurium(NCIM 2501),Staphylococcus aureus(NCIM 2122), Bacillus subtilis(NCIM 2193), and Pseudomonas aeruginosa(NCIM 2036).Nine out of 29 were found to be antibiotic producer.     

Analysis of cultivable aerobic bacterial community composition and screening for facultative sulfate-reducing bacteria in marine corrosive steel

Anaerobic, aerobic, and facultative bacteria are all present in corrosive environments. However, as previous studies to address corrosion in the marine environment have largely focused on anaerobic bacteria, limited attention has been paid to the composition and function of aerobic and facultative bacteria in this process. For analysis in this study, ten samples were collected from rust layers on steel plates that had been immersed in seawater for diff erent periods (i.e., six months and eight years) at Sanya and Xiamen, China. The cultivable aerobic bacterial community structure as well as the number of sulfate-reducing bacteria (SRB) were analyzed in both cases, while the proportion of facultative SRB among the isolated aerobic bacteria in each sample was also evaluated using a novel approach. Bacterial abundance results show that the proportions are related to sea location and immersion time;abundances of culturable aerobic bacteria (CAB) and SRB from Sanya were greater in most corrosion samples than those from Xiamen, and abundances of both bacterial groups were greater in samples immersed for six months than for eight years. A total of 213 isolates were obtained from all samples in terms of CAB community composition, and a phylogenetic analysis revealed that the taxa comprised four phyla and 31 genera. Bacterial species composition is related to marine location;the results show that Firmicutes and Proteobacteria were the dominant phyla, accounting for 98.13% of the total, while Bacillus and Vibrio were the dominant genera, accounting for 53.06% of the total. An additional sixfacultative SRB strains were also screened from the isolates obtained and were found to encompass the genus Vibrio (four strains), Staphylococcus (one strain), and Photobacterium (one strain). It is noteworthy that mentions of Photobacterium species have so far been absent from the literature, both in terms of its membership of the SRB group and its relationship to corrosion.     

三角褐指藻及其藻际细菌对不同无机氮源的响应

海洋生态系统中,浮游植物和细菌之间的相互作用是影响营养盐供应和再生、海洋初级生产力和生物地球化学循环的基本生态关系.为研究营养盐、浮游植物和细菌之间的关系,在实验室内培养条件下,通过改变氮源的形态(硝态氮和铵态氮)和浓度,研究海洋浮游植物三角褐指藻(Phaeodactylum tricornutum)的藻际细菌在不同营...     

海南红树林根系土壤中可培养细菌的多样性分析

为研究海南红树林根系土壤细菌多样性,充分发掘中国红树林特有微生物资源,作者于2012年12月份从10份采集于海南北港岛红树林根系淤泥样品中分离可培养海洋细菌,并基于16S rRNA基因序列分析的方法对样品中的细菌多样性进行研究。共分离得到122株海洋细菌,经并菌后选取其中的70株代表性菌株测序,结果发现它们分为4个类群:变形菌门(主要为γ-变形菌纲)(Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),分别占66%,19%,11%和4%。共代表30个属47个种,其中有9个潜在新种和2个潜在新属。本研究结果表明,北港岛红树林中存在着丰富的细菌多样性,很多类群可能是红树林特有细菌物种。     

海洋沉积物氨氧化菌群落结构的测序分析

本文以氨氧化细菌(ammonia-oxidizing bacteria,AOB)特征基因氨单加氧酶(ammonia monooxygenase,AMO)α亚基基因(amoA)为目标基因(～491bp),分别采用克隆文库、454高通量测序和illumina测序技术对其群落结构进行了比较研究。结果表明, 454高通量测序和illumina测序技术得到的菌群多样性及丰富度均高于克隆文库技术。在菌群组成方面,虽然3种测序技术检测到的优势类群均为亚硝化螺菌,但在菌群丰度方面存在差异。3种测序技术中,克隆文库技术在很大程度上可能高估物种丰度,且对部分低丰度类群的检测能力有限;illumina单端测序由于其测序片段较短,可能存在物种注释错误、不能准确区分各物种间序列差异等问题;比较而言, 454高通量测序技术能较为全面和准确地反映海洋沉积物中AOB的菌群结构特征。     

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。     

南、北极海洋不同种类细菌分离株中DMSP-依赖型脱甲基酶基因dmdA的系统发育多样性

Dimethylsulfoniopropionate(DMSP) is mainly produced by marine phytoplankton as an osmolyte, antioxidant,predator deterrent, or cryoprotectant. DMSP is also an important carbon and sulfur source for marine bacteria.Bacteria may metabolize DMSP via the demethylation pathway involving the DMSP demethylase gene(dmdA) or the cleavage pathway involving several different DMSP lyase genes. Most DMSP released into seawater is degraded by bacteria via demethylation. To test a hypothesis that the high gene frequency of dmdA among major marine taxa results in part from horizontal gene transfer(HGT) events, a total of thirty-one bacterial strains were isolated from Arctic Kongsfjorden seawater in this study. Analysis of 16S rRNA gene sequences showed that,except for strains BSw22118, BSw22131 and BSw22132 belonging to the genera Colwellia, Pseudomonas and Glaciecola, respectively, all bacteria fell into the genus Pseudoalteromonas. DmdA genes were detected in five distantly related bacterial strains, including four Arctic strains(Pseudoalteromonas sp. BSw22112, Colwellia sp.BSw22118, Pseudomonas sp. BSw22131 and Glaciecola sp. BSw22132) and one Antarctic strain(Roseicitreum antarcticum ZS2–28). Their dmdA genes showed significant similarities(97.7%–98.3%) to that of Ruegeria pomeroyi DSS–3, which was originally isolated from temperate coastal seawater. In addition, the sequence of the gene transfer agent(GTA) capsid protein gene(g5) detected in Antarctic strain ZS2–28 exhibited a genetically closely related to that of Ruegeria pomeroyi DSS–3. Among the five tested strains, only Pseudomonas sp. BSw22131 could grow using DMSP as the sole carbon source. The results of this study support the hypothesis of HGT for dmdA among taxonomically heterogeneous bacterioplankton, and suggest a wide distribution of functional gene(i.e., dmdA) in global marine environments.     

红树林海洋细菌的分离鉴定及其活性物质初步分析

从广西北部湾红树林海洋淤泥中分离筛选到270株海洋细菌,用喉癌细胞Hep-2的细胞毒作用为筛选模型,用SRB法检测这270株海洋细菌的抑制肿瘤细胞生长活性,其中13株呈阳性反应。依据形态和生理生化反应特征及16SrRNA基因序列分析,确定其中一株为短小芽孢杆菌Bacillus pumilusPLM4。取该菌株液体培养上清液,检测对人喉癌细胞Hep-2的生长抑制率为63.9%,经过凝胶过滤层析分析获得2个活性峰,其中活性高的峰估计平均分子量为116kd。采用经典显色反应法初步判断该活性物质为多糖。     

Ecotoxicological evaluation of sewage sludge using bioluminescent marine bacteria and rotifer

Bioassay using the marine bacteria,Vibrio fischeri and rotifer,Brachionus plicatilis, and chemical analyses were conducted to assess the toxicity of the various sewage sludges, one of the major ocean dumped materials in the Yellow Sea of Korea. Sludge elutriates extracted by filtered seawater were used to estimate the ecotoxicity of the sludge. Chemical characterization included the analyses of organic contents, heavy metals, and persistent organic pollutants in sludge. Bacterial bioluminescent inhibition (15 min), rotifer mortality (24 hr) and rotifer population growth inhibition (48 hr) assay were conducted to estimate the sludge toxicity. EC50 15 min (inhibition concentration of bioluminescence after 15 minutes exposed) values by Microtox® bioassay clearly revealed different toxicity levels depending on the sludge sources. Highest toxicity for the bacteria was found with the sludge extract from dyeing waste and followed by industrial waste, livestock waste, and leather processing waste. Clear toxic effects on the bacteria were not found in the sludge extract from filtration bed sludge and rural sewage sludge. Consistent with Microtox® results, rotifer neonate mortality and population growth inhibition test also showed highest toxicity in dyeing waste and low in filtration bed and rural sewage sludge. High concentrations of persistent organic pollutants (POPs) and heavy metals were measured in the samples from the industrial wastes, leather processing plant waste sludge, and urban sewage sludge. However, there was no significant correlation between pollutant concentration levels and the toxicity values of the sludge. This suggests that the ecotoxicity in addition to the chemical analyses of various sludge samples must be estimated before release of potential harmful waste in the natural environment as part of an ecological risk assessment.     

中国海洋生态效率时空格局演变及影响要素分析

随着海洋环境问题的日益凸显,海洋经济与海洋生态保护如何协调发展已成为学术界的研究热点。本文使用C2NLS方法对2003-2016年中国沿海11个省区市海洋生态效率进行测度,使用SDE方法对海洋生态效率的时空演进特征进行分析,最后对影响海洋生态效率的因素进行实证探索。结果表明：(1)海洋生态效率整体呈上升趋势,但地区差距并未显著缩小,海洋生态效率的地区分布出现极化趋势。(2)海洋生态效率重心表现出西南向移动特征,东北-西南向的空间格局更加明显。(3)环境规制、海洋科技和对外开放可以有效提高海洋生态效率水平;产业结构效应不显著,可能是由于高污染行业同时具有高产出与高排放的特征导致;区位因素效应不显著,海洋生态效率与地区发展水平之间没有明显的二次型关系,与中国海洋经济整体发展仍处于“倒U”型曲线的上升通道情况相符,部分印证了“环境的库兹涅茨”曲线理论。     

西印度洋产纤维素酶细菌的筛选及酶学特性初步研究

本研究从西印度洋表层沉积物中筛选获得1株高抗性、稳定性好的纤维素酶高产菌株XN149,通过形态学、革兰氏染色、生理生化鉴定以及16S rRNA基因序列分析对菌株进行分类鉴定,并研究了其酶学特性。结果表明,菌株XN149为漳州芽孢杆菌（Bacillus zhangzhouensis）,具有较高的产纤维素酶能力。酶学特性研究表明,该菌株所产纤维素酶的最适反应温度为65 ℃,最佳pH为6.0,该条件下发酵48 h后粗酶液酶活力为（60.19±0.31） U/mL。在30～60 ℃及pH为5.0～8.0时酶活性稳定性较高,Fe3+、Mn2+对该酶活性有一定的促进作用,Ba2+对其酶活性具有抑制作用。综上,漳州芽孢杆菌XN149是一株高产纤维素酶菌株,所产纤维素酶具有良好的耐热性和一定的酸碱耐受性,具有潜在的产业应用开发价值。     

培养基对海藻真菌PT2菌株的生长及活性代谢产物合成的影响

本文采用PD、GPY、YES、SYE、GPM5种不同培养基培养海藻真菌菌株PT2，并对其在不同培养基中的生长情况、代谢产物的形成及抗肿瘤活性进行分析，旨在了解不同培养基对菌株生长和活性物质合成的影响．结果显示，PT2菌株在不同培养基上的培养特征及生长速率有所不同，液体培养基发酵液抽提物的抗肿瘤活性也有较大差别，其中SYE发酵液抽提物活性最强，对KB、Raji和Hela 3种肿瘤细胞的IC50分别为7．6μg／cm^3、2．4μg／cm^3和3．3μg／cm^3．采用HPLC法对不同抽提物进行分析，发现它们之间的组成成分有显著差异．本文的实验结果表明，不同培养基对该菌株的生长及活性代谢产物的合成具有显著的影响。