地下水波动带中细菌群落结构与水质响应关系

为揭示地下水波动带中细菌群落结构特征及其与地下水环境相互作用关系,选取哈尔滨市第一水源地作为研究区,采集地下水样品以及波动带不同深度（0~5m非饱和带和6~50m饱和带）含水介质样品,分别用于水化学分析和16S rRNA细菌高通量测序,依托冗余分析定量表述地下水质参数与细菌群落相关性.水化学分析结果显示,研究区地下水主要污染物为Fe、Mn、NH₄⁺和有机质,Fe、Mn超标与研究区特定地质背景有关,NH₄⁺和有机质主要来源于人类活动.微生物分析结果显示,非饱和带和饱和带的细菌群落结构差异性显著,非饱和带细菌群落丰度和多样性显著高于饱和带,Proteobacteria、Bacteroidetes、Actinobacteria、Firmicutes和Acidobacteria为研究区优势门,在非饱和带和饱和带的累积相对丰度分别为82.89%和98.64%.冗余分析（RDA）结果显示,门水平上非饱和带中与水质演化强相关的细菌类群是Bacteroidetes、Proteobacteria、Actinobacteria、Verrucomicrobia,贡献率分别为15%、14.8%、8.9%和5.2%;饱和带中对地下水质演化起主要作用的类群为Bacteroidetes、Acidobacteria、Actinobacteria和Firmicutes,贡献率分别为38.4%、19.0%、10.8%和9.1%.属水平上非饱和带中的Pseudomonas和饱和带中的Flavobacterium对Fe、Mn、NH₄⁺生物转化起主导作用.本研究为揭示地下水波动带中生物地球化学作用对地下水环境的影响提供了科学依据,对地下水污染修复具有重要的意义.     

一株高效嗜碱反硝化菌的分离鉴定及其脱氮特性研究

为了应对日趋严苛的废水TN排放标准要求,试验采用溴百里酚蓝（BTB）培养基,从某煤化工废水处理厂反硝化缺氧池活性污泥中,经多次分离、纯化获得了一株高效兼性厌氧反硝化菌株HK13.通过形态观察及16S rRNA基因序列分析,菌株HK13被鉴定为施氏假单胞菌属（Pseudomonas stutzeri）.在此基础上,利用含硝酸盐模拟废水,探讨了碳源类型、C/N（碳氮比）、初始pH、溶解氧（以不同摇床转速表征不同浓度的溶解氧）和培养温度对菌株HK13反硝化脱氮能力的影响,确定了该菌株的最优生长条件和最大脱氮效率.结果表明:①菌株HK13最适反硝化脱氮条件为以柠檬酸钠碳源,C/N 8,培养温度35℃,初始pH 8~10,摇床转速100 r/min.②初始ρ（NO₃--N）为106.67 mg/L下,反应12 h后菌株HK13对TN的去除率可达92.62%;反应9~12 h时,该菌株的脱氮速率最高,可达20.03 mg/（L·h）,其16 h的脱氮率在98%以上,且无亚硝酸盐积累.③菌株HK13适宜生长的温度和pH范围广泛,分别为20~40℃和7~10.研究显示,菌株HK13具有快速高效的脱氮能力及嗜碱特性,拓宽了生物脱氮工艺对环境条件的适用范围,在废水脱氮领域具有广泛的应用前景.      

宁波三江口水域原核生物群落结构分析

首次应用16S rRNA基因-Illumina Miseq高通量测序技术对甬江流域宁波三江口区域的表层水体原核生物群落进行了分析,共获得215 504条高质量序列.多样性指数分析表明,该流域的水体原核生物群落具有较高的遗传多样性和丰富度.菌群分类分析发现,β-变形菌纲(β-Proteobacterium)、放线菌门(Actinobacteria)和拟杆菌门(Bacteroidetes)是主要的优势菌群,其相对丰度占总群落的78.88%.对比不同站位的原核生物群落,发现水文环境可能是影响三江口水域原核生物群落组成和结构的重要因素之一.与以往报道对比分析表明,甬江流域分布有多种污水和粪便污染指示菌,其中余姚江污染指示菌的丰度最高,说明其受污染的风险可能较高.病原菌BLASTN分析表明,在种和亚种水平上,分别检出76和18种潜在病原菌,占序列总量的2.19%和0.40%.本研究为系统认识甬江流域原核生物群落结构及生态功能提供了重要基础数据.     

Effect of reclaimed water effluent on bacterial community structure in the Typha angustifolia L. rhizosphere soil of urbanized riverside wetland, China

In order to evaluate the impact of reclaimed water on the ecology of bacterial communities in the Typha angustifolia L. rhizosphere soil,bacterial community structure was investigated using a combination of terminal restriction fragment length polymorphism and 16S rRNA gene clone library. The results revealed significant spatial variation of bacterial communities along the river from upstream and downstream. For example,a higher relative abundance of γ-Proteobacteria,Firmicutes,Chloroflexi and a lower proportion ofβ-Proteobacteria and ε-Proteobacteria was detected at the downstream site compared to the upstream site. Additionally,with an increase of the reclaimed water interference intensity,the rhizosphere bacterial community showed a decrease in taxon richness,evenness and diversity. The relative abundance of bacteria closely related to the resistant of heavy-metal was markedly increased,while the bacteria related for carbon/nitrogen/phosphorus/sulfur cycling wasn't strikingly changed. Besides that,the pathogenic bacteria markedly increased in the downstream rhizosphere soil since reclaimed water supplement,while the possible plant growth-promoting rhizobacteria obviously reduced in the downstream sediment.Together these data suggest cause and effect between reclaimed water input into the wetland,shift in bacterial communities through habitat change,and alteration of capacity for biogeochemical cycling of contaminants.     

玉门油田污染荒漠土壤石油降解菌多样性

为探索石油污染荒漠土壤石油降解微生物多样性、筛选高效石油降解菌,采用涂布平板法从石油污染荒漠土壤分离具有石油降解能力细菌,采用细菌形态观察和16S rRNA基因序列分析其多样性,并设计特异性引物,对分离细菌降解相关基因进行检测.结果表明,分离的37株细菌分别属于放线菌纲(Actinobacteria)、γ变形菌纲(Gammaproteobacteria)、β变形菌纲(Betaproteobacteria)、芽孢杆菌纲(Bacilli)和α变形菌纲(Alphaproteobacteria),分别占35.14%、32.43%、13.51%、13.51%、5.41%,归属于21个属的34个种类.优势菌属为假单胞菌属(Pseudomonas)、红球菌属(Rhodococcus)、微球菌属(Micrococcus)、寡养单胞菌属(Stenotrophomonas)、无色杆菌属(Achromobacter)和葡萄球菌属(Staphylococcus),占总数的51.35%,其中有36株细菌能以石油为唯一碳源稳定生长,对原油有明显的降解能力.在石油质量浓度为1 500 mg/L的基础培养基中,菌株YM43在培养7 d后对石油的降解率达55.47%,另有8株细菌的降解率不低于30.55%,11株细菌的降解率介于10.05%~28.37%,18株细菌的降解率不高于8.05%. PCR检测表明,有25株细菌含有烷烃单加氧酶基因,6株含芳烃双加氧酶基因,6株含联苯双加氧酶基因,4株含萘双加氧酶基因,3株含甲苯双加氧酶基因,2株含邻苯二酚双加氧酶基因.研究显示,石油污染荒漠土壤中可培养细菌具有高度多样性,分离的菌株有较强的石油降解能力,其降解功能与所存在的降解基因有关.      

垃圾填埋场中真细菌群落16S rRNA基因的分析

回灌式垃圾填埋场渗滤液中真细菌群落的多样性同样通过不依赖于微生物培养的分析而获得。利用特异性的引物对,选择性地扩增渗滤液DNA中的真细菌16S rRNA基因片断(16S rDNA),并用于构建16S rDNA克隆文库。文库内真细菌16S rDNA的遗传多样性通过限制片断长度多态性分析(RFLP,限制性内切酶Hin PII和Msp I)而获得。初步的结果表明,随机选出的200个真细菌克隆子被分为147个不同的RFLP型(组),当中丰度最高的两个型均仅含有9个克隆子,两者共占所有被分析克隆子的不到10%;克隆子数≥2的型共有21个(包括以上2个丰度最高的型),它们共代表74个克隆子,占所有被分析克隆子的37%;而剩下的126个型均只含有1个克隆子,它们共占整个基因文库的63%。由此可见,李坑垃圾渗滤液中的真细菌具有非常复杂的群落结构。     

Alga-lysing bioreactor and dominant bacteria strain

Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH4^＋-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time （HRT） of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% （in term of Chl-α） of coke reactor and filler reactor, respectively. And that of NH4^＋-N were 52.3% and 52.7%. The removal rates of CODMn were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.     

应用分子生物学与同位素示踪技术研究厌氧氨氧化菌活性及功效

厌氧氨氧化是一个新发现的氮循环途径,与反硝化作用都产生氮气,但准确衡量厌氧氨氧化活性及功效的相关方法还是个难题.本研究联合应用分子生物学与同位索示踪技术,以北京某污水处理厂的活性污泥为实验样品,研究其中厌氧氨氧化菌的存在、活性和功效.厌氧氨氧化菌的分子生物学分析是通过16S rRNA进行,选择两步引物分别为pla46-...     

Archaeal community structure along a gradient of petroleum contamination in saline-alkali soil

The impact of petroleum on archaeal community in salinealkali soils was investigated, which will expand the knowledge of the archaeal population involved in the natural attenuation of hydrocarbons in extreme environments.     

上海大气降水中细菌气溶胶的多样性研究

采用分子生物学技术16S rRNA克隆文库方法,以上海市2012年不同月份的大气降水样品为模板构建细菌16S rRNA克隆文库,对连续5个月的大气降水样品中细菌群落结构组成及多样性进行了研究,并对其群落结构的生物多样性、物种丰度及覆盖率等进行了分析比较.结果表明,变形菌门(Proteobacteria)(α-,β-,γ-)是上海市降水样品中细菌的优势菌群(32.5%~94.1%),另外还包括拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、异常球菌-栖热菌门(Deinococcus-Thermus)、蓝藻门(Cyanobacteria)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)共7个主要门类的细菌,以及未定菌(TM7).多样性指数分析结果显示,不同月份降水样品的细菌群落结构组成与多样性均存在着差异性,夏季6月样品的细菌群落生物多样性及物种丰度明显高于其它样品,其次是秋季8月的样品.