不同MBR反应器中硝化菌群落结构的研究

采用PCR-DGGE技术研究了正常运行工况下6个不同MBR反应器中的硝酸菌和反硝化菌的群落结构.结果表明,不同的MBR反应器好氧段中硝酸菌之间的相似性较高,相似性数值为78.7%~92.1%,但环境条件、进水水质及运行参数等因素的不同导致了不同MBR反应器硝酸菌群落的差异.MBR反应器中有5种硝酸菌和3种反硝化菌,其中优势硝酸菌种以α-变形细菌(Alpha proteobacteria)、生根瘤菌(Bradyrhizobium)、维氏硝酸菌(Nitrobacter winogradskyi)为主,反硝化菌以好氧反硝化菌-产碱杆菌属(Alcaligenes)、脱氮副球菌(Paracoccus denitrificans)、红球菌属(Rhodococcus)为主,说明反应器内同时存在好氧反硝化菌,推断MBR反应器内可能同时存在多种硝化和脱氮途径.     

Performance and role of N-acyl-homoserine lactone (AHL)-based quorum sensing (QS) in aerobic granules

The present study investigated the relationship between N-acyl-homoserine lactone（AHL）-based quorum sensing（QS） and the physico-chemical properties of aerobic granules.Stable mature granules were observed in SBR2 and SBR3 with average diameters of 0.96,and1.49 mm,respectively. The sludge densities of aerobic granules in SBR2 and SBR3 were1.0246,and 1.0201 g/mL,respectively,which were higher than that of flocculent sludge in SBR1（1.0065 g/mL）. The results showed that the activity of AHL-based QS in SBR2 and SBR3 amounted to 2.4- and 2.1-fold induction,however,that in SBR1 with flocculent sludge was1.6-fold induction. In addition,the results also showed that the activity of AHL-based QS in the three reactors rose in the feast condition,and then dropped with the consumption of substrate. However,the activity of AHL-based QS in these three reactors recovered again in prolonged starvation. Furthermore,the results showed that the enhancement of AHL-based QS favored the extracellular polymeric substance production of microorganisms in activated sludge. Thus,it could be concluded that aerobic granules showed higher AHL-based QS than flocculent sludge,which resulted from the higher sludge density of aerobic granules than flocculent sludge. AHL-based QS was related to the metabolism energy in the feast condition; however,in prolonged starvation,microorganisms would emit more AHL-like molecules to protect themselves to resist starvation. Moreover,the enhancement of AHL-based QS favored the EPS component productivity of the microorganisms in activated sludge,which contributed to maintain the aerobic granular structure.     

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).     

Effect of seed sludge on characteristics and microbial community of aerobic granular sludge

Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor.The influence of seed sludge on physical and chemical properties of granular sludge was studied;the microbial community structure was probed by using scanning electron microscope and polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE).The results showed that seed sludge played an important role on the formation of aerobic granules.Seed sludge taken from beer wastewater treatment plant(inoculum A) was more suitable for cultivating aerobic granules than that of sludge from municipal wastewater treatment plant(inoculum B).Cultivated with inoculum A, large amount of mature granules formed after 35 days operation, its SVI reached 32.75 mL/g, and SOUR of granular sludge was beyond 1.10 mg/(g·min).By contrast, it needed 56 days obtaining mature granules using inoculum B.DGGE profiles indicated that the dominant microbial species in mature granules were 18 and 11 OTU when inoculum A and B were respectively employed as seed sludge.The sequencing results suggested that dominant species in mature granules cultivated by inoculum A were Paracoccus sp., Devosia hwasunensi, Pseudoxanthomonas sp., while the dominant species were Lactococcus raffinolactis and Pseudomonas sp.in granules developed from inoculum B.     

Qualitatively and quantitatively assessing the aggregation ability of sludge during aerobic granulation process combined XDLVO theory with physicochemical properties

Inexact mechanism of aerobic granulation still impedes optimization and application of aerobic granules. In this study, the extended Derjaguin, Landau, Verwey, and Overbeek(XDLVO) theory and physicochemical properties were combined to assess the aggregation ability of sludge during aerobic granulation process qualitatively and quantitatively. Results show that relative hydrophobicity of sludge and polysaccharide content of extracellular polymeric substances(EPS) increased, while electronegativity of sludge decreased during acclimation phase. After 20 days' acclimation, small granules began to form due to high aggregation ability of sludge. Since then, coexisted flocs and granules possessed distinct physicochemical properties during granulation and maturation phase. The relative hydrophobicity decreased while electronegativity increased for flocs, whereas that for granules presented reverse trend. Through analyzing the interaction energy using the XDLVO theory, small granules tended to self-grow rather than self-aggregate or attach of flocs due to poor aggregation ability between flocs and granules during the granulation phase. Besides, remaining flocs were unlikely to self-aggregate owing to poor aggregation ability, low hydrophobicity and high electronegativity.     

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...     

硝化污泥保藏特性的研究

研究了不同保藏方法对硝化污泥硝化活性和相关性状的影响.结果表明,不同饥饿保藏条件下,各种方法下的硝化活性均呈指数衰减,自然环境、4 ℃、-20 ℃、-20℃加15%体积甘油方法下的活性衰减速率分别为0.012 d-1、0.021 d-1、0.019 d-1、0.013 d-1,半衰期分别为1.71、1.09、1.22、...     

同时产甲烷反硝化颗粒污泥中微生物群落结构

在UASB成功运行同时产甲烷反硝化小试基础上,针对反应器内颗粒污泥,通过构建古菌和细菌的16SrDNA基因文库、RADAR遗传变异分型和测序比对等技术进行了微生物系统发育关系和群落结构分析.结果表明,在颗粒污泥的古菌中,产甲烷髦毛菌和产甲烷杆菌是主要菌群,随机选出的88个古菌克隆子,这两类古菌的16SrDNA序列分别占古菌总量71.59%和22.73%;而颗粒污泥中细菌的多样性要高于古菌,低GC革兰氏阳性菌和ε变型菌纲分支的细菌是细菌的主要菌群,在随机选出的133个细菌克隆子中,这2类细菌的16SrDNA序列分别占细菌总量的49.62%和12.03%.     

河床原位修复对城市河流微生物群落的影响

采用分子生物学检测方法对实施了河床原位修复的河段进行全面的微生物群落分析。结果表明:河床生态修复后微生物群落结构及多样性发生了明显的变化。从微生物多样性指数来看,河床修复段的下游大于上游;从微生物群落结构来看,上游断面中ε-变形菌纲为最主要的细菌类型,多为致病病原菌,与该采样点周边人口密集而水质 经其他处理有关。经河床原位修复后,下游断面ε-变形菌纲丰度减少,α-变形菌纲和β-变形菌纲成为最优势的类群,其他细菌比例也相对均衡,NH₃-N和其他理化指标有所好转,说明微生物载体原位修复措施对城市景观河流黑臭的改善有一定的效果。     

Isolation and characterization of a strain with high microbial attachment in aerobic granular sludge

Aerobic granule is a special microbial aggregate associated with biofilm structure. The formation of aerobic granular sludge is primarily depending on its bacterial community and relevant microbiological properties. In this experiment, a strain with high microbial attachment was isolated from aerobic granular sludge, and the detailed characteristics were examined. Its high attachment ability could reach 2.34 (OD_600nm), while other low attachment values were only around 0.06-0.32, which indicated a big variation among the different bacteria. The strain exhibited a very special morphology with many fibric fingers under SEM observation. A distinctive behaviour was to form a spherical particle by themselves, which would be very beneficial for the formation and development of granular sludge. The EPS measurement showed that its PN content was higher than low attachment bacteria, and 3D-EEM confirmed that there were some different components. Based on the 16S rRNA analysis, it was identified to mostly belong to Stenotrophomonas. Its augmentation to particle sludge cultivation demonstrated that the strain could significantly promote the formation of aerobic granule. Conclusively, it was strongly suggested that it might be used as a good and potential model strain or chassis organism for the aerobic granular sludge formation and development.     

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.     

菌-藻共生好氧颗粒污泥的稳定性机理

在非曝气条件下接种好氧颗粒污泥(AGS)和绿藻藻种,经过18d的培养,成功构建了菌-藻共生好氧颗粒污泥系统(ABGS).研究表明,在非曝气条件下,与传统的AGS相比,ABGS具有更高的生物活性、除污染效能和机械强度,说明ABGS的稳定性更优.对ABGS的稳定性机理进行分析,发现在颗粒化过程中,胞外聚合物(EPS)特别是...     

Eubacteria and Archaea community of simultaneous methanogenesis and denitrification granular sludge

Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and community structure of microbes in the SMD granule in the UASB reactor were investigated by the construction of the Eubacteria and Archaea 16S rDNA clone libraries, fragment length polymorphism, and sequence blast. Real time quantitative-polymerase chain reaction (RTQ-PCR) technique was used to quantify the contents of Eubacteria and Archaea in the SMD granule. The contents of some special predominant methanogens were also investigated. The results indicated that the Methanosaeta and Methanobacteria were the predominant methanogens in all Archaea in the SMD granule, with contents of 71. 59% and 22. 73% in all 88 random Archaea clones, respectively. The diversity of Eubacteria was much more complex than that of Archaea. The low GC positive gram bacteria and Б-Protebacteria were the main predominant Eubacteria species in SMD granule, their contents were 49. 62% and 12. 03% in all 133 random Eubacteria clones respectively. The results of RTQ-PCR indicated that the content of Archaea was less than Eubacteria, the Archaea content in total microorganisms in SMD granule was about 27. 6%.     

受四环素影响的活性污泥胞内外聚合物特征

利用离心加超声的方法对处理四环素抗生素废水的活性污泥胞外聚合物(EPS)和胞内聚合物(IPS)进行提取,将其分为-溶解型胞外聚合物(slime)、松散附着型胞外聚合物(LB)、紧密附着型胞外聚合物(TB)、胞内聚合物(IPS).用BCA法和蒽酮-硫酸法测定了各层EPS和IPS溶液中的蛋白质和多糖含量,同时用三维荧光光谱扫描(EEM)和同步荧光光谱扫描(SFS)对其中的溶解性有机物(DOM)进行定性和定量分析,结果表明在EPS中的蛋白质和多糖的含量高于IPS,且蛋白质含量均比多糖含量高,而在slime和IPS中的蛋白质和多糖的含量高于LB和TB;低激发波长类色氨酸和高激发波长类色氨酸是EPS和IPS溶液的主要成分,且溶液中的溶解性代谢产物和芳香蛋白类物质相对含量在slime、LB、TB和IPS逐渐增加,而难降解有机物-腐殖酸类物质的相对含量逐渐降低;进水四环素浓度与污泥各层EPS和IPS溶液中DOM也具有较好的相关性;三维荧光法检测溶液中的DOM与化学分析测试出的蛋白质含量具有较好的相关性.     

处理含油废水的好氧颗粒污泥形成过程及其特性研究

含油废水中因含有较高浓度的油脂类物质和聚合物而对环境造成危害,威胁人类健康,同时,为解决采用传统膜分离工艺运行成本较高的难题,开展了基于好氧颗粒污泥技术的含油废水处理研究.结果表明,以含油废水启动反应器,经35 d好氧颗粒污泥培养成熟,COD和溶解性油的去除率高达86.0%和94.2%;在絮状污泥颗粒化过程中,污泥胞外聚合物中蛋白质类物质含量提高3.7倍,蛋白质类物质与多糖类物质比值升高到2.72,证明胞外聚合物内蛋白质类物质浓度增加是活性污泥颗粒化的重要因素;好氧颗粒污泥荧光光谱结果显示好氧颗粒污泥中蛋白质类物质的稳定存在是好氧颗粒污泥形成的重要因素.选取好氧颗粒污泥技术处理含油废水的效果和成本均优于常规生物处理工艺和膜分离工艺,由于污泥及其胞外聚合物中多糖类和蛋白质类物质含量均较高,适用于回收污泥资源,对含油污泥的资源化利用意义重大.     

信号分子在好氧颗粒污泥形成过程中的作用

采用人工配水成功培养好氧颗粒污泥,针对絮状污泥颗粒化过程中污泥的理化性质、污染物去除效率、胞外聚合物和信号分子变化进行相关分析.结果发现,在好氧颗粒污泥的形成过程中,污泥对污染物去除能力明显提高.与接种污泥相比,成熟的好氧颗粒污泥对COD、NH₄⁺-N和PO₄^3--P去除率分别提高20%、36%和57%.好氧颗粒污泥中胞外蛋白质和多糖含量分别增加了116和31mg/gMLVSS.采用激光共聚焦扫描显微镜对接种污泥和好氧颗粒污泥进行空间表征,发现后者蛋白质含量明显增加,说明蛋白质在污泥颗粒化过程中具有重要作用;磷酸二酯酶活性总体呈现上升趋势,第二信使环二鸟苷酸（cyclic diguanylic,c-di-GMP）含量先增加后降低,与胞外聚合物中蛋白质和多糖的变化规律相似.通过SPSS软件进一步分析得知,c-di-GMP与蛋白质相关系数R²=0.92871,呈极显著相关性;与紧密结合型胞外聚合物中蛋白质相关系数R²=0.89025,呈显著相关性,推测c-di-GMP可能是通过指导紧密结合型胞外聚合物中的蛋白质合成以促进好氧颗粒污泥的形成.     

Effect of wastewater COD/N ratio on aerobic nitrifying sludge granulation and microbial population shift

The effect of COD/N ratio on the granulation process and microbial population succession was investigated. Four identical sequencing batch reactors, R1, R2, R3 and R4, were operated with various initial COD/N ratios ranging from 0/200 to 800/200 (m/m). Ethanol was fed as the source of COD. Aerobic granules were successfully cultivated in R2 and R3, operating with the COD/N ratio of 200/200 and 400/200, respectively. Scanning electron microscope observations indicated that short rod-shaped and spherical bacteria were dominant in R2, while granules produced in R3 were surrounded with a large amount of filamentous bacteria. The average specific nitritation rate in R2 and R3 were 0.019 and 0.008 mg N/(mg MLVSS.hr), respectively. Fluorescence in situ hybridization results demonstrated that nitrifying bacteria population was enriched remarkably in R2. It indicated that nitrification ability and nitrifying bacteria population were enriched remarkably at low COD/N ratio. However, no granules were formed in R1 and R4 which might attribute to either limited or excessive extracellular polymeric substances production. This study contributed to a better understanding of the role of COD/N ratio in nitrifying sludge granulation.     

连续进水对好氧颗粒污泥稳定维持的影响

为了考察连续进水对好氧颗粒污泥稳定维持的影响,在SBR反应器中接种成熟的好氧颗粒污泥,采用连续进水、间歇出水的方式运行.在反应器运行过程中,部分好氧颗粒污泥解体,完整存在的好氧颗粒污泥表面附着生长树枝状污泥和少量丝状菌.SVI在最初的13d内逐步上升,随后趋于稳定.同时,考察了好氧颗粒污泥在连续进水运行过程中EPS含量的变化以及EPS的空间分布变化:胞外多糖的含量在运行过程中呈下降趋势;胞外蛋白的含量整体呈先上升后下降的趋势;反应器运行15d后,胞外蛋白和胞外多糖的含量趋于稳定.连续进水运行30d后,EPS空间分布最大变化是β-D吡喃葡萄糖,它大量聚集在颗粒的表层及颗粒中伸出的树枝状污泥及丝状菌的表层.本研究表明,连续进水的运行方式不利于好氧颗粒污泥的稳定维持,底物浓度梯度为好氧颗粒污泥稳定维持的重要因素之一;EPS的分泌量以及空间分布对好氧颗粒污泥的形成及其稳定维持起着至关重要的作用.     

有机物特性对AAO系统污泥沉降性能的影响

研究了不同进水混合型有机物组成下AAO系统中活性污泥菌群结构演替规律以及微生物胞内、胞外聚合物的变化对污泥沉降性能的影响.结果表明,当进水中有机物全部为溶解态时,污泥沉降性能最佳,污泥体积指数(sludge volume index,SVI)为70 m L·g-1,且优于进水有机物以溶解态为主(SVI=120 m L·g-1)和以颗粒态为主(SVI=280 m L·g-1)的系统.根据菌群结构分析可知Thiothrix、Chryseolinea和Trichococcus这3种菌属对污泥沉降性能的影响至关重要.其中颗粒态有机物可促进Trichococcus的生长,而溶解态有机物可促进Thiothrix和Chryseolinea的生长.此外,菌群结构的改变也对胞内及胞外聚合物的变化有重要影响,从而加剧污泥沉降性能改善或恶化的进程.较高的溶解态有机物含量可提高胞内聚合物贮存能力,并改善污泥沉降性能.同时,污泥沉降性能也与松散附着胞外聚合物(loosely bound extracellular polymeric substances,LB-EPS)中多糖、蛋白质和Zeta电位呈显著负相关关系.     

微塑料对变形杆菌生物膜生长发育的影响

微塑料对生物膜的影响现阶段主要集中于对成熟生物膜群落组成结构变化的研究,缺乏对不同发育阶段生物膜的研究.为探究微塑料对不同发育阶段生物膜的影响,选用环境中广泛存在的变形杆菌和1 μm聚苯乙烯微塑料(PS-MPs)作为研究对象,通过暴露实验研究PS-MPs对生物膜生物量、胞外聚合物(EPS)组成和胞外酶活性的影响.结果表...