Temporal and spatial changes of microbial community in an industrial effluent receiving area in Hangzhou Bay

Anthropogenic activities usually contaminate water environments, and have led to the eutrophication of many estuaries and shifts in microbial communities. In this study, the temporal and spatial changes of the microbial community in an industrial effluent receiving area in Hangzhou Bay were investigated by 454 pyrosequencing. The bacterial community showed higher richness and biodiversity than the archaeal community in all sediments. Proteobacteria dominated in the bacterial communities of all the samples; Marine_Group_Ⅰ and Methanomicrobia were the two dominant archaeal classes in the effluent receiving area. PCoA and AMOVA revealed strong seasonal but minor spatial changes in both bacterial and archaeal communities in the sediments. The seasonal changes of the bacterial community were less significant than those of the archaeal community, which mainly consisted of fluctuations in abundance of a large proportion of longstanding species rather than the appearance and disappearance of major archaeal species. Temperature was found to positively correlate with the dominant bacteria, Betaproteobacteria, and negatively correlate with the dominant archaea,Marine_Group_Ⅰ; and might be the primary driving force for the seasonal variation of the microbial community.     

悬浮填料对厌氧氨氧化MBR运行的影响特性及机理

向厌氧氨氧化（anammox）膜生物反应器（MBR）投加悬浮填料,考察其对反应器脱氮性能和膜污染的影响特性,并探究了相关机理.试验结果表明,投加填料后,反应器脱氮性能良好.当进水氨氮（NH₄⁺-N）160mg/L、亚硝态氮（NO₂^--N）180mg/L时,出水NH₄⁺-N和NO₂^--N均在15mg/L以下,硝态氮（NO₃^--N）在30mg/L以下,总氮去除率可达90%.投加填料显著减轻了膜污染,跨膜压差（TMP）稳定在8kPa左右.混合液中溶解性微生物产物（SMP）和胞外聚合物（EPS）成分分析结果表明,在第67~149d,蛋白质总量、多糖总量和总有机碳总量分别下降了49%、43%和61%,它们浓度的下降有利于延缓膜污染;此外,悬浮填料对膜组件的机械碰撞也起到了物理清洗作用.高通量测序结果显示,悬浮填料生物膜在anammox菌相对丰度方面显著高于混合液污泥,说明anammox菌更适宜于附着生长,投加填料可以为其提供更加稳定的生长环境.     

污泥性质对微波预处理-厌氧消化的影响及古菌群落结构分析

污泥性质对预处理强化厌氧消化工艺的效果有较大影响.本研究对比了某实际污水厂A~2O工艺和A~2O-MBR工艺产生的剩余污泥在微波预处理-厌氧消化过程中污泥性质的变化与产气效果,并考察了在预处理和厌氧消化过程中污泥的古菌群落结构变化.结果表明,A~2O工艺剩余污泥有机质含量比A~2O-MBR污泥高出16. 4%(分别为66. 4%和50. 0%),SCOD、溶解性蛋白质和多糖分别为后者1. 24、2. 02和4. 84倍,具有更好的可生物降解性.虽然预处理对生物降解性差的A~2O-MBR污泥有机物释放效果更好,但A~2O污泥在微波预处理-厌氧消化后产甲烷量比相应处理后的A~2O-MBR污泥多26. 1%.两种剩余污泥的古菌群落结构差异较大,A~2O-MBR污泥中甲烷丝菌属和甲烷八叠球菌属丰度分别比A~2O污泥多3. 68%和19. 73%.预处理对古菌群落的丰富度和均匀度影响相对较小,但厌氧消化后波动较大.污泥中有机组分不同是引起古菌群落结构变化的重要影响因素.     

膜生物反应器系统中原生动物的群落特征

与常规生物处理工艺相比,膜生物反应器(MBR)系统由于污泥停留时间长、污泥浓度高等特点,因此反应器内微生物种群结构呈现不同的特点.通过对MBR系统中原生动物的群落结构进行为期近2年连续的监测结果表明,过长的污泥龄对系统中原生动物的种类与数量均有负面的影响.研究发现:①在长期不排泥的MBR系统中,当环境温度从11℃上升到25℃,原生动物的多样性呈显著上升趋势;②当污泥龄从350 d下降到30 d,MBR系统中原生动物的种类与数量明显增加.特别在较低的环境温度条件下,系统中原生动物的种类与数量随着污泥龄的缩短呈显著的上升趋势;③与相同污泥龄(SRT=30d)的氧化沟(TOD)系统相比,MBR的污泥中原生动物的种类与数量均低于TOD.     

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.     

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...     

MFC-MBR耦合系统中SMP与EPS特性的研究

将膜生物反应器与以剩余污泥为底物的微生物燃料电池结合建立了能够有效控制膜污染的MFC-MBR耦合系统,以传统MBR作为对照研究了MFC-MBR耦合系统内泥水混合液的溶解性微生物产物(SMP)与胞外聚合物(EPS)浓度、分子量分布及荧光特性的变化情况.结果表明与传统MBR相比耦合系统中SMP与TB-EPS浓度分别升高了8.5%与9.2%,而LB-EPS浓度则降低了47.9%,并且LB-EPS中芳香性蛋白质与腐植酸的浓度也显著降低.同时还考察了传统MBR与MFC-MBR耦合系统中膜污染情况,结果发现,在传统MBR中TMP达到30KPa所需时间为38d,而MFC-MBR耦合系统则为55d.说明MFC污泥回流能够有效减缓膜污染的发生.     

南海东北部珠江口盆地沉积物中古菌群落垂向分布研究

本文利用高通量测序及荧光定量PCR技术对南海东北部珠江口盆地柱状沉积物的表层（0~1 cm）、中层（20~21 cm）和底层（40~41 cm）3个样品中古菌群落结构及其垂向分布进行了研究。结果表明,沉积物中的古菌主要可被划分为奇古菌门、Woesearchaeota、深古菌门和广古菌门。表层样品中的古菌群落多样性和丰富度最低,类群Marine Group I（MGI）占绝对优势,中层和底层样品中的古菌则以MGI、Group C3、Marine Benthic Group B、Thermoplasmata为主要类群。执行氨氧化功能的亚硝化侏儒菌属（Nitrosopumilus）是研究区域的优势类群,预示该区域的氮代谢过程较为活跃。定量结果表明,古菌丰度为2.00×107~8.20×107copies/g,且随深度增加呈下降趋势。研究结果可为进一步探讨南海沉积物中的古菌群落特征提供依据。     

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.     

Monitoring biofouling based on aerobic respiration in reverse osmosis system

A monitoring method of biofouling in reverse osmosis (RO) system was proposed based on the fluorescent signal of resorufin, which is reduced by nicotinamide adenine dinucleotide released from viable cells during aerobic respiration. The fluorescent signal of resorufin reduced by planktonic cells and microorganisms of biofilm showed linearity, indicating its feasibility to monitor biofouling in a RO system. For the application of the method to the lab-scale RO system, the injection concentration of resazurin and the injection flow rate were optimized. Biofilm on RO membranes continuously operated in a lab-scale RO system was estimated by resorufin fluorescence under optimized detection condition. As a result, resorufin fluorescence on RO membrane showed a significant increase in which the permeability of RO system decreased by 30.48%. Moreover, it represented the development of biofilm as much as conventional biofilm parameters such as adenosine triphosphate, extracellular polymeric substances, and biofilm thickness. The proposed method could be used as a sensitive and low-cost technology to monitor biofouling without autopsy of membranes.     

PAC强化MBR工艺处理效果的研究进展

膜生物反应器（MBR）是将膜分离技术与活性污泥法相结合而形成的废水处理工艺。由于MBR具有较高的去除率使其被广泛地应用于废水处理中,但是膜堵塞和膜污染问题依旧是阻碍MBR工艺进一步推广的主要原因。颗粒活性炭（PAC）具有高的比表面积,其较强的吸附能力可有效地缓解MBR工艺运行过程中膜堵塞和膜污染问题。通过总结近年来对膜碳生物反应器（PAC-MBR）的研究进展,分析了影响MBR工艺的影响因素（pH、HRT、SRT、曝气量、SS）,综述了PAC-MBR联合工艺对废水中氨氮、BOD₅、COD、TC、TN、浊度的处理效果。     

Bioaugmentation with a pyridine-degrading bacterium in a membrane bioreactor treating pharmaceutical wastewater

The bacterial strain Paracoccus denitrificans W12, which could utilize pyridine as its sole source of carbon and nitrogen, was added into a membrane bioreactor （MBR） to enhance the treatment of a pharmaceutical wastewater. The treatment efliciencies investigated showed that the removal of chemical oxygen demand, total nitrogen, and total phosphorus were similar between bioaugmented and non-bioaugmented MBRs, however, significant removal of pyridine was obtained in the bioaugmented reactor. When the hydraulic retention time was 60 hr and the influent concentration of pyridine was 250-500 mg/L, the mean effluent concentration of pyridine without adding W12 was 57.2 mg/L, while the pyridine was degraded to an average of 10.2 mg/L with addition of W12. The bacterial community structure of activated sludge during the bioaugmented treatment was analyzed using polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE）. The results showed that the W12 inoculum reversed the decline of microbial community diversity, however, the similarity between bacterial community structure of the original sludge and that of the sludge after bioaugmentation decreased steadily during the wastewater treatment. Sequencing of the DNA recovered from DGGE gel indicated that sp., Sphingobium sp., Comamonas sp., and Hyphomicrobium sp. were the dominant organisms in time sequence in the bacterial community in the bioaugmented MBR. This implied that the bioaugmentation was affected by the adjustment of whole bacterial community structure in the inhospitable environment, rather than being due solely to the degradation performance of the bacterium added.     

Biofilm structure and its influence on clogging in drip irrigation emitters distributing reclaimed wastewater

Using reclaimed wastewater for crop irrigation is a practical alternative to discharge wastewater treatment plant effluents into surface waters.However,biofouling has been identified as a major contributor to emitter clogging in drip irrigation systems distributing reclaimed wastewater.Little is known about the biofilm structure and its influence on clogging in the drip emitter flow path.This study was first to investigate the microbial characteristics of mature biofilms present in the emitters and the effect of flow path structures on the biofilm microbial communities.The analysis of biofilm matrix structure using a scanning electron microscopy (SEM) revealed that particles in the matrix of the biofilm coupled extracellular polysaccharides (EPS) and formed sediment in the emitter flow path.Analysis of biofilm mass including protein,polysaccharide and phospholipid fatty acids (PLFAs) showed that emitter flow path style influenced biofilm community structure and diversity.The correlations of biofilm biomass and discharge reduction after 360 h irrigation were computed and suggest that PFLAs provide the best correlation coefficient.Comparatively,the emitter with the unsymmetrical dentate structure and shorter flow path (Emitter C) had the best anti-clogging capability.By optimizing the dentate structure,the internal flow pattern within the flow path could be enhanced as an important method to control the biofilm within emitter flow path.This study established electron microscope techniques and biochemical microbial analysis methods that may provide a framework for future emitter biofilm studies.     

基因工程菌生物强化处理系统微生物群落分析

在膜-生物反应器(MBR)和传统活性污泥反应器(CAS)中,考察了基因程菌生物强化对阿特拉津的去除效果,并通过PCRDGGE分析了反应器不同运行阶段污泥微生物群落的变化.结果表明,基因工程菌生物强化实现了阿特拉津的高效生物去除,MBR和CAS阿特拉津平均去除率分别达到88.6％和85.3％.阿特拉津生物强化去除有助于反...     

Molecular analysis of long-term biofilm formation on PVC and cast iron surfaces in drinking water distribution system

To understand the impacts of different plumbing materials on long-term biofilm formation in water supply system, we analyzed microbial community compositions in the bulk water and biofilms on faucets with two different materials-polyvinyl chloride （PVC） and cast iron, which have been frequently used for more thanlO years. Pyrosequencing was employed to describe both bacterial and eukaryotic microbial compositions. Bacterial communities in the bulk water and biofilm samples were significantly different from each other. Specific bacterial populations colonized on the surface of different materials. Hyphomicrobia and corrosion associated bacteria, such as Acidithiobacillus spp., Aquabacterium spp., Limnobacter thiooxidans, and Thiocapsa spp., were the most dominant bacteria identified in the PVC and cast iron biofilms, respectively, suggesting that bacterial colonization on the material surfaces was selective. Mycobacteria and Legionella spp. were common potential pathogenic bacteria occurred in the biofilm samples, but their abundance was different in the two biofilm bacterial communities. In contrast, the biofilm samples showed more similar eukaryotic communities than the bulk water. Notably, potential pathogenic fungi, i.e., Aspergillus spp. and Candida parapsilosis, occurred in similar abundance in both biofilms. These results indicated that microbial community, especially bacterial composition was remarkably affected by the different pipe materials （PVC and cast iron）.     

反硝化聚磷菌快速富集、培养及其荧光原位杂交技术鉴别

以污水处理厂A2/O厌氧段污泥为种泥,采用膜生物反应器(membrane bio-reactor,MBR)对反硝化聚磷菌(denitrifyingphosphate-removal bacteria,DPB)进行快速富集及培养,并提供一种鉴别方法.试验中以乙酸钠为碳源,并在缺氧段投加一定浓度的硝酸盐,结果表明,在膜组件的高效截留作用下,经过厌氧-好氧和厌氧-缺氧2个阶段的富集培养,35 d内反硝化聚磷菌占全部聚磷菌(phosphate-accumulating organisms)的比例从24%上升到93%.此时系统的脱氮、除磷效率均可保持在90%以上.通过荧光原位杂交技术(fluorescence in situ hybridization,FISH)并结合常规测定手段对活性污泥进行鉴别,确定Pseudomonas sp.和Rhodocyclus sp.为主要的优势菌.     

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.      

MBR与SMBR脱氮除磷特性及膜污染控制

为提高污水深度处理效能和工艺运行的稳定性,研究以序批式膜生物反应器(SMBR)与传统膜生物反应器(MBR)为对象,对比研究其脱氮除磷特性、缺氧时间对工艺效率的影响及膜污染控制策略,同时应用分子生物学技术对两种工艺中微生物群落结构和组成进行分析.结果表明,间歇曝气能强化系统脱氮,使SMBR工艺去除总氮效果优于MBR,而在氨氮、总磷、COD、浊度去除方面两者无明显差异,去除率分别为94%、78%、80%、97%.延长SMBR工艺缺氧时间对COD、氨氮去除无显著影响,降低了总氮、总磷的去除率,总氮去除率由61%下降到46%,总磷由74%下降到52%.采用间歇曝气和投加一定浓度的粉末活性炭(PAC)均有利于减缓膜污染.微生物群落分析发现,两种工艺中微生物群落结构和组成无显著差异,硝化螺菌属(Nitrospira)和脱氯单胞菌属(Dechloromonas)为系统中的高丰度功能菌群,为工艺高效运行提供了生物学基础.     

SMBR工艺处理废水时操作方式的研究

膜生物反应器(MBR)在废水处理中必须能保证长期稳定运行,而操作方式与MBR运行性能密切相关。通过对小试规模的浸没式MBR(SMBR)恒流和恒压两种操作方式的短期以及长期实际运行研究,发现次临界区恒流操作和经济操作压力TMP以下的恒压操作有利于MBR的长期稳定运行;并从理论上分析了对应于不同的操作区域的膜污染机理。     

响应土壤阴离子类型的盐碱土古细菌群落多样性研究

为初步解析我国黑龙江苏打盐碱地（HA）、新疆荒漠盐碱地（XD）、山西平原盐碱地（SY）、江苏滨海盐碱地（JD）和天津滨海盐碱地（TB）5个地区的土壤古细菌群落结构多样性,以及其响应土壤不同阴离子类型的分布特征,采用基于Illumin-Hiseq平台的高通量测序技术获得489~604个古细菌可操纵分类单元（OTU）.OTU多样性分析结果表明XD的古细菌群落物种丰富度最高,SY和HA分别具有最高和最低的古细菌群落多样性和均匀度;OTU的物种注释结果表明HA以奇古菌门（Thaumarchaeota）为优势菌门,其他土壤均以广古菌（Euryarchaeota）为优势;综合5个样品古细菌群落结构、优势古细菌属和土壤阴离子进行的典范对应分析（CCA）结果显示,XD古细菌群落结构和Haloterrigena属（XD中最优势,相对丰度20.2%）明显的响应土壤SO₄^2-浓度;HA古细菌群落结构和Nitrososphaera属（HA中最优势,62.3%）明显的响应土壤HCO₃^-/CO₃^2-浓度和pH;JD、SY和TB古细菌群落结构、Halorubrum属（JD和TB中最优势,24.4%和15.6%）,和Natronomonas属（SY和JD中第二优势,8.1%和9.2%）明显的响应土壤Cl^-浓度.研究结果说明我国不同地域分布的盐碱土受其土壤阴离子类型及其浓度的影响,呈现不同的古细菌群落结构和分布特征.研究结果为我国不同类型盐碱土古细菌资源的挖掘提供依据,也为揭示我国不同类型盐碱土古细菌群落的生态功能提供参考.