处理不同废水MBR系统中微生物群落结构的比较

为了研究膜生物反应器中微生物群落结构与系统处理效能的关系.为工艺改进提供依据,从4种处理不同水质的MBR污泥中提取细菌总基因组DNA,采用PCR-DGGE和克隆测序技术对系统中的微生物群落结构进行了解析,根据序列数据进行同源性分析并建立了系统发育树.结果表明,在长期稳定运行后不同MBR中形成了各自特有的生态群落,进水水质对总细菌的群落结构有着较大的影响,处理含有较复杂成分废水的反应器中,种群多样性较高,Shannon指数分别为0.77和0.78.总细菌中,主要优势种群以Proteobacteria纲(8个OUTs)和Bacillus属(2个OUTs)为主.不同反应器中氨氧化菌群结构较为相似,存在着相同的顶级优势群落;测序结果表明MBR中存在着多种亚硝化菌属,其中以亚硝化单胞菌属最为普遍,并鉴定出2株反硝化菌属UncuItured Achromobacter sp.和Uncultured denitrifying bacterium,说明反应器中可能同时存在多种硝化和脱氮途径.     

利用PCR-DGGE研究膜生物反应器中微生物的群落结构

使用聚合酶链式反应-变性梯度凝胶电泳技术(PCR-DGGE)考查了天津某再生水处理厂膜生物反应器(Membrane Bioreactor,简称MBR)培养驯化直至正常运行全过程中总细菌群落结构的演替情况.结果表明,在MBR环境中,接种的传统活性污泥中的微生物群落在几天内发生了较大的变化,在进污水驯化时,微生物群落也遭受了冲击,最后经过培养驯化趋于稳定,一些菌种逐渐成长为顶级优势微生物,在反应器内占据主导地位.最终该反应器逐渐形成了自己独有的微生物群落生态系统.另外,对该微生物群落的部分优势总细菌进行了克隆测序和系统发育树分析,通过鉴定获得的10条总细菌的16S rDNA序列,它们分别与气单胞菌属、假单胞菌、亚硝酸菌属、丛毛单胞菌属和杆菌的同源性在97%以上,这些优势微生物在MBR反应器去除有机物的过程中起到了关键的作用.     

生物膜MBR反应器和MBR反应器处理洗涤废水比较

比较生物膜MBR反应器和MBR反应器处理洗涤废水的效果。结果表明,两个系统对COD、LAS及氨氮的去除均具有良好的处理效果。和MBR反应器相比,生物膜MBR反应器的运行条件要好。生物膜MBR反应器的运行条件:水力停留时间(HRT)4～4.5h,气水比351∶,而MBR反应器的运行条件:水力停留时间(HRT)9～10h,气水比451∶。通过两个反应器抗冲击负荷实验的研究,结果表明,在进水水质相同的条件下,就膜生物反应器的上清液而言,生物膜MBR反应器具有更好的抗冲击负荷能力。     

MBR处理腈纶废水的效能及微生物群落结构分析

采用序批式膜生物反应器(SBMBR)处理腈纶聚合废水和丙烯腈废水,考察了不同废水比例和运行条件下反应器的运行效果,并采用PCR-DGGE技术分析了反应器内微生物群落结构的变化.结果表明,SBMBR对腈纶废水具有较好的处理效果,碳源和碱度不足是影响脱氮效果的两个主要因素;在水力停留时间为24 h,90 min缺氧/150 min好氧交替运行的条件下,COD、NH+4-N和TN的平均去除率分别为82.5%、98.7%和74.6%,出水水质可以稳定达到国家污水综合排放标准(GB8978-1996)的一级标准;PCR-DGGE分析结果表明,反应器内微生物多样性与进水水质和运行条件密切相关,微生物群落呈现出较明显的演替过程;通过克隆测序,从9个污泥样品中成功鉴定获得22种微生物的16S r DNA序列,并筛选出7种降解腈纶废水的优势菌种.     

MBR中微生物演变及其对膜污染影响的试验研究

膜污染是制约膜生物反应器(MBR)应用的重要因素,对膜污染机理以及防治方法进行研究具有重要的现实意义。试验对MBR中微型动物演变及其对膜污染的影响进行了分析。随运行时间的延长MBR中优势种群依次为游动型纤毛虫、累枝虫和钟虫、表壳虫、红斑瓢体虫。优势种群对污泥状态具有指示作用,而污泥状态对膜污染有着直接影响,污泥状态恶化时膜污染速度约为污泥状态良好时的6倍。     

不同污泥龄膜生物反应器内微生物的群落结构特征

为研究不同污泥龄MBR(膜生物反应器)内的微生物群落结构特征,构建了SRT(污泥龄)分别为10、20、40和80 d的4个平行的MBR,通过PCR-DGGE技术获得各MBR内微生物的DNA指纹图谱,并对条带进行切胶测序. 结果表明,SRT不影响MBR对废水的处理效果,不同SRT下MBR对NH₄+-N和COD_Cr的去除率均能达到90%以上. DGGE结果表明,在运行过程中,各MBR内的微生物群落结构均发生了明显变化,并且不同SRT的MBR内总细菌群落结构变化特征相同;SRT影响同一微生物在各MBR中的出现速率;总细菌Shannon-Wiener多样性指数和丰富度指数均随着SRT的增加而升高,SRT为80 d的MBR内Shannon-Wiener多样性指数最高. 测序结果表明,不同SRT的MBR内的优势种属不同,其中Arcobacter sp.、beta proteobacterium及Thiothrix sp.为不同SRT的MBR中共同存在的关键菌属,对MBR的运行起着重要作用.      

利用PCR-DGGE技术分析生物陶粒硝化反应器中微生物群落动态

为了揭示生物陶粒硝化反应器中活性污泥的微生物种群多样性,从运行不同时期的反应器中提取活性污泥,利用PCR-DGGE技术初步分析了生物陶粒反应器细菌的种群演替情况.结果表明,随着进水COD值的逐阶段降低,氨氮去除率逐步提高到64.38%.群落结构和优势种群的数量具有时序动态性,微生物多样性与废水的处理效果出现协同变化的特征.测序结果表明,生物陶粒反应器中运行第21天的优势种群除了自养细菌,还有异养细菌存在.其中自养细菌为Nitrosospira.sp和Nitrobacter.sp,异养细菌为Bacillus.sp、Pseudomonas.sp和Pseudochrobactrum.sp.     

MBR中微生物群落结构的演变与分析

为了揭示膜.生物反应器中微生物群落结构多样性的演变过程,通过细胞裂解法直接提取不同时期污泥中的基因组DNA,利用基于16SrDNA的PCR-DGGE技术获得了微生物群落的DNA特征指纹图谱,并对条带进行了统计分析和切胶测序,使用序列数据进行了同源性分析并建立了系统发育树.DGGE分析表明,在反应器运行前17d内污泥中微生物群落结构变化很大,与接种污泥的相似性系数下降到了29.2%,从而说明MBR中处理工艺和进水水质的改变导致微生物群落结构多样性降低.在试验过程中,Pscudomonas和Aeromonas hydrophila等种群一直保持着较为稳定的优势地位,也有原始种群如Bacillus sp.的消亡和以Enterococcus faecalis、Comamonas sp.、Fusobacterium sp.等为代表的次级种群的强化和演变.UPGMA聚类分析将DGGE图谱区分为3大类群并对应于各自的运行时期.测序结果表明,MBR中微生物菌群间进化距离较大,其中Proteobaeteria纲和Bacillus属细菌较多.在反应器运行后期演变为优势地位的菌群(如Comamonas sp.)加剧了膜污染物的产生和积累.     

MBR和CAS工艺污泥在贫营养培养条件下的微生物群落结构研究

采用聚合酶链式-变性梯度凝胶电泳（PCR-DGGE）技术,研究了膜生物反应器（MBR）和传统活性污泥工艺（CAS）反应器中微生物在贫营养条件下的总细菌群落结构.结果表明,在培养过程中,污泥的微生物种群经历了一个比较明显的变化过程,且以CAS污泥微生物种群的变化更为明显,演替过程中既有原始优势种群的消亡,又有新的优势种群...     

膜-生物反应器污泥缺氧反硝化过程中SMP的形成

在不同C/N比下,研究了膜-生物反应器在缺氧状态下溶解性微生物产物(SMP)的形成规律.结果表明,不同C/N比的反硝化过程,随着底物的降解,SMP都呈现不同程度的累积,且高C/N比下的合成量(9~10mgCOD/gVSS)高于低C/N比下的合成量(3~4mgCOD/gVSS).从SMP的组分来看,不同C/N比下糖类的代谢规律相似,蛋白质则随着C/N比的升高其含量占SMP总量的比例增加,成为膜污染潜在的主要贡献者.随着反硝化过程的进行,体系中亚硝酸盐的累积对SMP的形成没有影响.     

不同DO下MBR内微生物群落结构与运行效果关系

应用A/O-MBR处理实际生活污水,考察了不同溶解氧(DO)条件下,微生物群落结构与其处理效果的对应关系.结果表明,DO浓度在0.2~4.0mg/L对COD去除效果无明显影响,COD平均去除率均在90%以上.DO浓度变化对NH4+-N去除影响较大,DO浓度下降到0.2mg/L时,NH4+-N平均去除率由99%下降到65%.通过PCR-DGGE分析,较高DO条件下(4.0,2.0mg/L)的总细菌微生物群落多样性高于较低DO条件(0.5,0.2mg/L),但其群落结构变化与与反应器的处理效果对应关系不明显;氨氧化菌的群落结构变化较明显,且在不同的DO条件下起主要作用的氨氧化菌的菌属不同,其群落变化与反应器的NH4+-N去除效果相对应,DO为2.0,0.5mg/L时氨氧化菌群落结构比较相似,此时反应系统的脱氮效果也比较好.     

Shifts in microbial community structure and diversity in a MBR combined with worm reactors treating synthetic wastewater

The chemical oxygen demand (COD) and NH₃-N removal, membrane fouling, sludge characteristics and microbial community structure in a membrane bioreactor (MBR) coupled with worm reactors (SSBWR) were evaluated for 210 days. The obtained results were compared to those from a conventional MBR (C-MBR) operated in parallel. The results indicated that the combined MBR (S-MBR) achieved higher COD and NH₃-N removal efficiency, slower increase in membrane fouling, better sludge settleability and higher activities of the related enzymes in the activated sludge. Denaturing gradient gel electrophoresis was used to analyze the microbial community structures in the C-MBR and the S-MBR. The microbial community structure in the S-MBR was more diverse than that in the C-MBR. Additionally, the slow-growing microbes such as Saprospiraceae, Actinomyces, Frankia, Clostridium, Comamonas, Pseudomonas, Dechloromonas and Flavobacterium were enriched in the S-MBR, further accounting for the sludge reduction, membrane fouling alleviation and wastewater treatment.     

高密度微阵列基因芯片技术在微生物分子生态学研究中的运用

应用基于16S rDNA 的高密度微阵列基因芯片(Microarray)技术对膜生物反应器内的微生物多样性进行研究.结果表明,膜生物反应器具有较高的微生物多样性,Microarray共检测到1 019种微生物.Proteobacteria是其中的优势种,共含有657种微生物,占总种群数量的64.5%左右.gamma Proteobacteria是Proteobacteria中的优势种,占35.8%左右,但相对含量一般并不高.beta Proteobacteria虽然种群数量稍少,但其在荧光强度最高的前25种和50种微生物中比重最大,分别占40%和36%左右.通过比较微生物的相对荧光强度,发现Clostridia是系统中的优势微生物种属.一些常见的硝化细菌如Nitrosomonadaceae、Nitrospiraceae等也具有较高的含量.Microarray作为一种实时、高效、准确的分子生物学手段,可应用于废水处理中的微生物多样性研究.     

Isolation and characterization of heterotrophic nitrifying bacteria in MBR

The study presented the method for isolating the heterotrophic nitrifiers and the characterization of heterotrophic nitrification. Continuous tests via a membrane bioreactor (MBR) were operated under the controlled conditions to proliferate the nitrifiers. Heterotrophic nitrifying bacteria were isolated from the system in which the efficiency of total nitrogen(TN) removal was up to 80%. Since no autotrophic ammonium and nitrite oxidizers could be detected by fluorescence in situ hybridization(FISH), oxidized-N production was unlikely to be catalyzed by autotrophic nitrifiers during the heterotrophic nitrifiers‘ isolation in this study. The batch test results indicate that the isolated heterotrophic bacteria were able to nitrify. After 3 weeks incubation, the efficiencies of the COD removal by the three isolated bacterial strains B1,B2 and B3 were 52.6%, 71.7%, and 77.7%, respectively. The efficiencies of the TN removal by B1, B2, and B3 were 35.6%, 61.2% and 68.7%, respectively.     

Influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules

In order to evaluate the influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules, these granules were cultivated with different seed sludge, and the variation of microbial community and dominant bacterial groups that impact the nitrogen removal efficiency of the aerobic nitrifying granules were analyzed and identified using 16s rDNA sequence and denaturing gradient gel electrophoresis (DGGE) profiles. The results presented here demonstrated that the influence of the community structure of seed sludge on the properties of aerobic nitrifying granules was remarkable, and the granules cultivated by activated sludge from a beer wastewater treatment plant showed better performance, with a stable sludge volume index (SVI) value of 20 mL/g, high extracellular polymeric substance (EPS) content of 183.3 mg/L, high NH₄⁺-N removal rate of 89.42% and abundant microbial population with 10 dominant bacterial groups. This indicated that activated sludge with abundant communities is suitable for use as seed sludge in culturing aerobic nitrifying granules.     

不同好氧颗粒污泥中微生物群落结构特点

为了探讨活性污泥好氧颗粒化过程对微生物种群的影响、不同底物及不同颗粒化方法培养的好氧颗粒污泥中微生物群落结构的差异,以接种污泥、模拟废水好氧颗粒污泥和分别投加粉末活性炭和硅藻土的实际生活污水好氧颗粒污泥为研究对象,利用PCR-DGGE对比分析了接种污泥和好氧颗粒污泥中的微生物群落结构.结果表明:活性污泥好氧颗粒化过程会减少微生物种群多样性,影响颗粒污泥稳定性的细菌被淘汰,而聚磷菌、反硝化菌、难降解有机物降解菌等污水处理功能微生物都在颗粒化过程中得到保留.活性污泥好氧颗粒化过程中能够实现亚硝化细菌(AOB)一定程度的富集.与接种活性污泥相比,好氧颗粒污泥中AOB的多样性指数与均匀性指数均有提高.好氧颗粒污泥中的优势菌群主要分布于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和未培养菌(uncultured bacterium).其中AOB均属于β-Proteobacteria的亚硝化单胞菌属(Nitrosomonas).     

Assessing the impact of fungicide enostroburin application on bacterial community in wheat phyllosphere

Fungicides have been used extensively for controlling fungal pathogens of plants. However, little is known regarding the effects that fungicides upon the indigenous bacterial communities within the plant phyllosphere. The aims of this study were to assess the impact of fungicide enostroburin upon bacterial communities in wheat phyllosphere. Culture-independent methodologies of 16S rDNA clone library and 16S rDNA directed polymerase chain reaction with denaturing gradient gel electrophoresis (PCR-DGGE) were used for monitoring the change of bacterial community. The 16S rDNA clone library and PCR-DGGE analysis both confirmed the microbial community of wheat plant phyllosphere were predominantly of the γ-Proteobacteria phyla. Results from PCR-DGGE analysis indicated a significant change in bacterial community structure within the phyllosphere following fungicide enostroburin application. Bands sequenced within control cultures were predominantly of Pseudomonas genus, but those bands sequenced in the treated samples were predominantly strains of Pantoea genus and Pseudomonas genus. Of interest was the appearance of two DGGE bands following fungicide treatment, one of which had sequence similarities (98%) to Pantoea sp. which might be a competitor of plant pathogens. This study revealed the wheat phyllosphere bacterial community composition and a shift in the bacterial community following fungicide enostroburin application.     

Evaluation of factors influencing soluble microbial product in submerged MBR through hybrid ASM model

In this study, a mathematical model was established to predict the formation of the soluble microbial product (SMP) in a submerged membrane bioreactor. The developed model was calibrated under the reference condition. Simulation results were in good agreement with the measured results under the reference condition. The calibrated model was then used in the scenario studies to evaluate the effect of three chosen operating parameters: hydraulic retention time (HRT), dissolved oxygen concentration, and sludge retention time (SRT). Simulation results revealed that the SMP dominated the soluble organic substances in the supernatant. The scenario studies also revealed that the HRT can be decreased to 1 h without deteriorating the effluent quality; dissolved oxygen concentration in the reactor can be kept at 2–3 mg/L to maintain the effluent quality, reduce the content of SMP, and minimize operating costs; the optimal SRT can be controlled to 10–15 d to achieve complete nitrification process, less membrane fouling potential, and acceptable organic removal efficiency.