Effect of Changing VOC Influent Composition on the Microbial Community Structure of TBABs

Microbial communities in trickle bed air biofilters (TBABs) were evaluated under conditions of interchanging the feed volatile organic compounds (VOCs) and VOC mixtures. Three independent TBABs (Biofilter “A,” “B,” and “C”) were run under interchanging VOCs conditions with different initial VOCs. Two aromatic compounds (toluene and styrene) and two oxygenated compounds (methyl ethyl ketone (MEK) and methyl isobutyl ketone (MIBK)) were interchanged as single solutes. Two other TBABs “D” and “E” were run for two VOC mixtures. Biofilter “D” had a VOC mixture with equal molar ratio of the four components and Biofilter “E” received a VOC mixture with its composition based on EPA 2003 emission report. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes was used to assess the microbial richness in TBABs for treating the VOC mixtures and the impact of interchanging VOCs on the bacterial community structure in the biofilters. The results from DGGE indicated that the microbial community structure in the biofilter was different after each interchange of VOCs. Some bands of microbial species faded and some bands were strengthened. For the two TBABs treating VOC mixtures, the microbial species did not show significant difference, but the richness among these species was different from each other.     

城市污水强化生物除磷工艺中除磷菌种群结构分析

采用PCR-DGGE等分子生物学技术。从平行AN／AO工艺六个反应池的活性污泥样品中提取总DNA，采用套式PCR（nested PCR）进行两轮PCR扩增。对α-Proteobaeteria、β-Proteobacteria的16S rDNA片段进行DGGE（变性梯度凝胶电泳）分离，从而分析活性污泥样品中该类除磷菌的种群结构。结合测序技术并通过NCBI（美国国立生物技术信息中心）基因库比对，证实生物除磷过程中存在着丰富的α-Proteobacteria、β-Proteobacteria，同时可以得到其中优势类群的初步认识。结果表明，PCR-DGGE结舍测序技术可以用于污水处理过程中样品的除磷菌群落结构分析。     

污泥干化芦苇床中芦苇内生菌群多样性研究

以污泥干化芦苇床中芦苇内生菌的多样性作为主要研究对象,结合16S rDNA高通量测序技术与PCR-DGGE技术,对污泥干化芦苇床和天然湿地中污泥菌群及芦苇内生菌群多样性进行了比较分析。结果表明,污泥干化芦苇床芦苇内生菌的细菌类群包含了20门、30纲、73目、152科、467属、1 167种细菌。其中变形菌门(65.6%)和拟杆菌门(25.5%)占绝对优势,γ-变形菌纲是丰度最高的纲,占总数的43.4%;排前20位的优势菌种相对百分比之和为46.5%,分属于所对应的18个属、12科、10目、6纲、3门。DGGE分析表明,污泥干化芦苇床中的污泥和芦苇内生菌优势菌群的种类与天然湿地相比差异显著;同时由于污泥的影响,芦苇内生菌群种类也发生了较为明显的变化。     

固态发酵过程中微生物总DNA提取方法比较

为了分析固态发酵过程中微生物群落的多样性及演替情况,对比研究了从固态发酵中提取细菌和真菌DNA的3种方法--溶壁酶法、超声波法、液氮研磨 CTAB法.使用紫外分光光度计测定了由不同提取方法得到的DNA的产量与纯度;使用细菌16S rDNA基因通用引物(341F和907R)和真菌18S rDNA基因通用引物(NU-SSU-0817和NU-SSU-119)对DNA进行了PCR扩增;采用DGGE(变性梯度凝胶电泳)法对固态发酵中细菌和真菌的多样性进行了分析.结果显示,3种方法得到的粗提和纯化DNA长度均约为23 kb;细菌和真菌PCR产物长度分别约为586 bp和422 bp.细菌和真菌PCR产物的DGGE分析表明,3种方法提取的DNA所反映的微生物多样性比较一致;但紫外分光光度计测定结果表明溶壁酶法提取固态发酵中微生物总DNA产量最高,超声波法次之,液氮研磨 CTAB法最低.     

Effects of Cd（Ⅱ） and Cu（Ⅱ） on microbial characteristics in 2-chlorophenol-degradation anaerobic bioreactors

The effects of Cd^2＋ and Cu^2＋ at 300 mg/L on anaerobic microbial communities that degrade 2-cholorophenol （2-CP） were examined. Based on the polymerase chain reaction （PCR） of 16S rDNA, bacterial community diversity and archaeal community structure were analyzed with denaturing gradient gel electrophoresis （DGGE） and cloning, respectively. Degradation capabilities of the anaerobic microbial community were drastically abated and the degradation efficiency of 2-CP was reduced to 60% after shock by Cu^2＋ and Cd^2＋, respectively. The bacterial community structure was disturbed and the biodiversity was reduced after shock by Cu^2＋ and Cd^2＋ for 3 d. Some new metal-resistant microbes which could cope with the new condition appeared. The sequence analysis showed that there existed common Archaea species in control sludge and systems when treated with Cu^2＋ and Cd^2＋, such as Methanothrix soehngenii, Methanosaeta concilii, uncultured euryarchaeote, and so on. Both the abundance and diversity of archaeal species were altered with addition of Cd^2＋ and Cu^2＋ at high concentration. Although the abundance of the predominant archaeal species decreased with Cd^2＋ and Cu^2＋ addition for 3 d, they recovered to some extent after 10 d. The diversity of archaeal species was remarkably reduced after recovery for 10 d and the shift in archaeal composition seemed to be irreversible. The 2-CP-degradation anaerobic system was more sensitive to Cu^2＋ than Cd^2＋.     

PCR-DGGE技术对城市餐厨垃圾堆肥中细菌种群结构分析

使用基于16S rDNA的PCR-DGGE(变性梯度凝胶电泳)技术对城市餐厨垃圾堆肥过程中细菌种群结构随时间的变化进行了研究.在堆肥不同时间取样,进行了堆肥温度、pH、含水率、有机质、C/N的变化分析和DGGE分析.结果显示,堆肥温度高于50℃的天数为7d,最高达65℃,可有效杀灭致病菌;最终pH值接近7.5;C/N为20.04.PCR-DGGE图谱显示,不同时间堆肥样中细菌DGGE图谱有着明显的差异性;堆肥升温期细菌种群丰富,优势种群不明显;高温期细菌种群减少,优势种群明显;降温期细菌种群结构基本保持稳定.温度对堆肥过程中细菌种群具有明显的筛选作用.堆肥各阶段DGGE图谱相似性Cs值比较低,堆肥处理后细菌种群结构与堆肥原料之间存在明显差异.     

沉积物再悬浮对浮游细菌群落结构的影响

本研究通过小宇宙的模拟实验,设置3组处理,即无再悬浮的对照组(C)、强(SR)和弱(WR)2个不同再悬浮强度的处理组,采用末端限制性片段长度多态性(terminal restriction fragment length polymorphism,T-RFLP)和变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)分析不同处理组中浮游细菌群落结构的变化.结果表明,再悬浮处理组浮游细菌的物种丰度高于对照组,并且弱再悬浮中浮游细菌物种丰度高于强再悬浮处理.弱再悬浮中细菌多样性也高于强再悬浮处理.浮游细菌与环境因子的典型对应分析(CCA)和冗余分析(RDA)结果表明,浮游动物枝角类(Cladocera)生物量和颗粒态磷(PP)浓度与浮游细菌群落结构动态变化均表现出显著的相关性(p0.05).由此可见,沉积物再悬浮对浅水湖泊浮游细菌的种类和多样性存在重要影响,其可能通过影响浮游动物群落结构以及水体中营养盐的形态和浓度对浮游细菌群落产生影响.     

接种纤维素降解菌对牛粪堆肥微生物群落的影响

以牛粪和稻草为堆肥原料,利用传统培养法和PCR-DGGE技术相结合对接种纤维素降解菌后堆肥的微生物群落的动态变化进行了分析.结果表明,接种菌堆肥与自然堆肥相比,微生物数量的变化趋势快于自然堆肥,接种菌在堆肥初期能够激发微生物增殖,快速启动堆肥发酵,缩短堆肥进程.接种菌堆肥比自然堆肥的DGGE凝胶图谱条带的差异性大,接种...     

原油降解中微生物群落的16S rDNA-PCR-DGGE分析

为对原油降解过程中微生物群落结构的变化,本文采用直接向原油中加入营养物刺激的方法,获得原油降解菌体系。提取不同培养时间条件下降解菌体系的总DNA,并以之为模板采用PCR-DGGE技术研究了在原油降解过程中微生物群落的组成,结果表明,随着培养时间的不同,微生物的群落结构表现出差异,随着时间的变化其微生物群落也会发生改变。回收了DGGE图谱中的18个条带,测序结果经过Blast比对表明其中10个条带代表的细菌是不可培养的,显示了DGGE技术的优越性。同时采用气相色谱技术对降解过程中的原油烃的变化进行了分析,结果显示原油烃类组分会随着降解菌多样性的变化而发生变化,降解菌体系表现出了其对烷烃类物质的降解能力。     

农业废物好氧堆肥过程因子对细菌群落结构的影响

采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术研究了农业废物堆肥过程中细菌种群随时间的变化.同时,应用Quantity One2.0和Canoco4.5软件对获得的堆肥细菌种群数据与堆肥过程因子:环境温度、堆体温度、pH、含水率、水溶性有机碳(WSC)、C/N、水溶性氨氮(NH4+-N)和硝氮(NO3--N)进行冗余分析,并做出样点、种群与堆肥过程因子的二维排序图.结果表明,细菌群落(样点)以堆肥过程因子为梯度大体可划分为升温期(1～2d)、高温期(3～11d)、降温期(12～18d)和腐熟期(19～36d)4个阶段,每一阶段均有对应种群存在.不同的堆肥过程因子对细菌种群的影响大小依次为:NO3--N堆体温度WSCC/NNH4+-N含水率pH环境温度,其中,堆体温度、WSC、NO3--N、NH4+-N对细菌种群的影响极显著(p0.01),C/N、pH对细菌种群的影响显著(p0.05),含水率、环境温度对细菌种群的影响不显著.