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%.     

废水同步脱硫脱氮关键工艺参数及微生物群落结构的研究

大量未经处理的含硫化物和硝酸盐废水的排放将带来严重的环境问题.根据以废治废原则,使用厌氧滴滤塔反应器构建的同步脱硫耦联反硝化脱氮反应(SDD)能很好的去除废水中S~(2-)和NO-x-N.其中以聚氨酯泡沫为填料的厌氧滴滤塔反应器中生物活性最强,脱氮脱硫效果最好.体系中功能菌优先将S~(2-)氧化成S0,待S~(2-)去除完全后,再进一步将S0氧化成SO_4~(2-).同时,SDD反应降解NO_3~--N的速率快于NO_2~--N.进水S/N摩尔比越大,产物中SO_4~(2-)相对含量越低.结合实际工程考虑,应控制进水S/N摩尔比在5/3~5/2之间,S~(2-)浓度控制在538 mg·L-1以下.微生物群落结构分析结果表明,Thiobacillus属在4组反应器上占绝对优势,其相对丰度均高于40%.其次相对丰度较高的Rhodanobacter、Arenimonas和Truepera属与厌氧反硝化作用密切相关.对4组反应器中微生物进行Alpha-多样性分析结果表明取得较好脱硫耦联反硝化效果的体系中物种多样性指数也较高.     

Amoxicillin effects on functional microbial community and spread of antibiotic resistance genes in amoxicillin manufacture wastewater treatment system

This study aimed to reveal how amoxicillin(AMX) affected the microbial community and the spread mechanism of antibiotic resistance genes(ARGs) in the AMX manufacture wastewater treatment system. For this purpose, a 1.47 L expanded granular sludge bed(EGSB) reactor was designed and run for 241 days treating artificial AMX manufacture wastewater. 454 pyrosequencing was applied to analyze functional microorganisms in the system. The antibiotic genes OXA_(-1), OXA_(-2), OXA_(-10), TEM_(-1), CTX-M_(-1), class I integrons(intI1) and 16 SrRNA genes were also examined in sludge samples. The results showed that the genera Ignavibacterium, Phocoenobacter,Spirochaeta, Aminobacterium and Cloacibacillus contributed to the degradation of different organic compounds(such as various sugars and amines). And the relative quantification of eachβ-lactam resistance gene in the study was changed with the increasing of AMX concentration.Furthermore the vertical gene transfer was the main driver for the spread of ARGs rather than horizontal transfer pathways in the system.     

Microbial community functional structure in response to micro-aerobic conditions in sulfate-reducing sulfur-producing bioreactor

Limited oxygen supply to anaerobic wastewater treatment systems had been demonstrated as an effective strategy to improve elemental sulfur(S0) recovery, coupling sulfate reduction and sulfide oxidation. However, little is known about the impact of dissolved oxygen(DO) on the microbial functional structures in these systems. We used a high throughput tool(GeoChip) to evaluate the microbial community structures in a biological desulfurization reactor under micro-aerobic conditions(DO: 0.02–0.33 mg/L). The results indicated that the microbial community functional compositions and structures were dramatically altered with elevated DO levels. The abundances of dsrA/B genes involved in sulfate reduction processes significantly decreased(p 0.05, LSD test) at relatively high DO concentration(DO: 0.33 mg/L). The abundances of sox and fccA/B genes involved in sulfur/sulfide oxidation processes significantly increased(p 0.05, LSD test) in low DO concentration conditions(DO: 0.09 mg/L) and then gradually decreased with continuously elevated DO levels. Their abundances coincided with the change of sulfate removal efficiencies and elemental sulfur(S0) conversion efficiencies in the bioreactor. In addition, the abundance of carbon degradation genes increased with the raising of DO levels, showing that the heterotrophic microorganisms(e.g., fermentative microorganisms) were thriving under micro-aerobic condition. This study provides new insights into the impacts of micro-aerobic conditions on the microbial functional structure of sulfatereducing sulfur-producing bioreactors, and revealed the potential linkage between functional microbial communities and reactor performance.     

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%.     

不同接种污泥的厌氧氨氧化反应器启动特性及菌群结构演替规律分析

采用两套相同的膨胀颗粒污泥床(EGSB),分别接种城市污水处理厂活性污泥(A)和厌氧颗粒污泥(B)启动厌氧氨氧化反应器,考察了A、B反应器中污泥的形态、脱氮性能、容积氮负荷和氮去除负荷的差异,同时利用高通量测序技术从分子生物学水平分析启动过程中菌群结构演替规律.结果表明:反应器运行119 d后,A、B反应器均成功培养出成熟的厌氧氨氧化颗粒污泥,其中相比于B反应器,A反应器缩短了厌氧氨氧化启动过程的菌体自溶和活性迟滞阶段.微生物群落结构分析结果进一步表明,在活性迟滞和活性提高阶段,A反应器中浮霉菌门(Planctomycetes)的丰度分别为1.18%、5.98%,而B颗粒污泥反应器中浮霉菌门(Planctomycetes)的丰度仅为0.92%、1.41%,说明A反应器Planctomycetes菌在菌体自溶和活性迟滞阶的增长速度大于B反应器.此外,经过119 d的启动,A反应器中Candidatus Brocadia丰度可以达到11.34%,而B反应器中Candidatus Brocadia丰度仅为7.28%.     

同步脱氮好氧颗粒污泥的特性及其反应过程

厌氧颗粒污泥经过驯化后,成为具有同步硝化与反硝化(SND)功能的好氧颗粒污泥.实验在2L反应器中进行,温度,pH值,溶解氧分别控制在25℃,pH7～8,3～4mg/L.在实验条件下,SND好氧颗粒污泥COD去除率90%,氨氮去除率100%,出水检测不出NO₂^--N和NO₃^--N.反应器中SND颗粒污泥粒径在2.0～2.7mm的占全部颗粒污泥的50%,SVI为15～30mL/gTSS;污泥所能承受的最大压力为23.236N;SND好氧颗粒污泥中挥发性固体为9.92mg/mL,占总固体的2/3.采用SND好氧颗粒污泥进行脱氮研究,反应6h后氨氮去除率达100%,废水中检测不到NO₂^--N,仅残留2mg/L的NO₃^--N.     

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.     

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).     

不同结构芳香酸对厌氧颗粒污泥特性与微生物群落的影响

以苯甲酸(BA)、邻苯二甲酸(PA)、连苯三甲酸(HA)、1-萘甲酸(1NA)为研究对象,探究了不同结构芳香酸对厌氧颗粒污泥理化特性与微生物群落的影响.结果表明,在40 d的接触实验中,1NA实验组对溶解性化学需氧量(SCOD)的去除率为86.09％,与空白对照组相比降低了7％.4个实验组污泥疏松胞外聚合物(LB-EP...     

反硝化脱硫工艺中微生物群落结构及动态分析

为了研究反硝化脱硫工艺(denitrifying sulfide removal,DSR)中微生物群落与工艺运行的相关性,实验提取了反应器运行不同阶段污泥样品中微生物的全基因组DNA,利用高通量宏基因组学技术———基因芯片来解析各阶段功能微生物群落的结构特征及其演替过程.通过对功能基因的Simpson、Shannon多样性指数和聚类分析表明,微生物群落结构会随着反应器运行的阶段进行相应调整,且变化较大.在运行的前两个阶段,由于不适应环境,微生物群落的多样性指数比起始时明显减少,随着反应器的运行污泥逐渐成熟,多样性指数迅速增加,也标志着反应器进入稳定运行阶段.通过对反硝化脱硫过程中关键基因相对丰度的分析发现,功能基因的丰度不仅能反映功能菌群的活性,而且与工艺处理效果密切相关.     

Bamboo charcoal addition enhanced the nitrogen removal of anammox granular sludge with COD: Performance, physicochemical characteristics and microbial community

The effects of different chemical oxygen demand(COD) concentrations on the anammox granular sludge with Bamboo Charcoal(BC) addition were evaluated in UASB reactor. The results showed that the average total nitrogen(TN) removal efficiency was reduced from85.9% to 81.4% when COD concentration was increased from 50 to 150 mg/L. However, the TN removal efficiency of BC addition reactors was dramatically 3.1%–6.4% higher than that without BC under different COD concentrations. The average diameter o...     

微电解耦合同步硝化反硝化工艺强化煤化工废水脱氮效能及微生物群落结构研究

研究探讨微电解耦合同步硝化反硝化(ICME-SND)工艺对煤化工废水氮去除的强化效能及机理.结果表明,在ICME-SND工艺中,煤化工废水的COD、氨氮和总氮的去除率分别为85.99％、85.81％和75.59％,SND效率达到83.06％,显著高于非强化SND工艺.同时,ICME-SND工艺中活性污泥与微电解反应释放...     

上升流速对CANON工艺稳定性及微生物群落的影响

采用膨胀颗粒污泥床(EGSB)反应器作为全程自养脱氮(CANON)工艺启动运行的装置,考察了不同上升流速对CANON工艺脱氮性能的影响,并对固定生物膜-活性污泥(IFAS)系统内颗粒污泥粒径的变化和生物膜上的生物量进行定量分析,同时对颗粒污泥和生物膜上的微生物进行高通量分析,探究在不同聚集体上微生物群落结构的特点.结果...     

生物-电协同驱动型同步脱氮除硫工艺性能及功能菌群分析

同步脱氮除硫燃料电池可同时去除氮硫污染物并产生电能,其同步脱氮除硫功能受到生物驱动和电能驱动的双重作用.为探究该新型工艺中生物驱动和电能驱动的贡献,分别构建了电能驱动型、生物驱动型和生物-电驱动型同步脱氮除硫系统,并考察了其基质去除性能、产物类型分布和电能产生情况.结果表明,生物驱动型和生物-电驱动型系统皆具有良好的同步脱氮除硫功能,而电能驱动系统仅具有硫氧化功能.通过衡算不同驱动型系统对基质去除的贡献率以及对产物产生的贡献比,发现在生物-电驱动系统中,电能驱动和生物驱动在硝酸盐还原,硫酸盐和氮气的生成以及电量产生方面皆具有协同效应.并通过高通量测序分别从门和属分类水平对生物驱动系统和生物-电驱动系统阳极室内污泥细菌群落进行分析,相对丰度较高的菌门都是Proteobacteria和Bacteroidetes,而生物驱动系统的优势菌属为Halothiobacillus和Thauera,生物-电驱动系统的优势菌属为Marinilabiaceae和Rhodanobacter.     

长期储存亚硝化颗粒污泥的活化及菌群结构变化

采用无机人工配水,通过逐级提高进水氨氮负荷(0.32~0.64kg/(m³·d))和设定合适的初始游离氨浓度(3.7~7.2mg/L),在SBR反应器中对常温(24~29℃)下储存1a的亚硝化颗粒污泥(NGS)进行了活化,并使用Miseq高通量测序技术分析了污泥中微生物多样性的变化情况.结果表明,NGS的亚硝化性能可在短时间内恢复.运行8d后,反应器的氨氮去除率达到95%以上,亚硝态氮累积率超过了80%,但污泥粒径持续减小,胞外聚合物(EPS)含量明显降低.活化至第20d,NGS的氨氮比去除速率和亚硝态氮比累积速率分别达到24.6mg/(gVSS·h)、23.8mg/(gVSS·h),平均粒径稳定在0.5mm左右.在活化期间,绝大部分厌氧、异养菌属被洗脱,污泥的微生物多样性显著降低.Nitrosomonas等氨氧化菌的相对丰度由活化前的1%上升至约58%,同时,Nitrospira等硝化菌的生长受到了选择性抑制.这意味着即使经历长期的常温储存,NGS仍可作为SBR的接种污泥,实现反应器的快速启动.     

Long-term impacts of graphene oxide and Ag nanoparticles on anammox process: Performance, microbial community and toxic mechanism

The increasing application of engineered nanoparticles (NPs) has posed an emerging challenge to constructed wetland wastewater treatment. The performance, microbial community and toxic mechanism of anammox-based unplanted subsurface-flow constructed wetlands (USFCWs) were investigated under the long-term exposure of different graphene oxides (GOs) and Ag NP concentrations. Results showed that the addition of GO could promote TN removal, manifesting as function anammox bacteria C. Anammoxoglobus having a relative high abundance, for GO did not cause significant damage to the cell integrity though there was an increase in ROS concentrations. TN removal would not be obviously affected under exposure of 1?mg/L Ag NPs, for the function gene related to cell biogenesis and repair was up-regulated; while the addition of 10?mg/L Ag NPs would have an inhibiting effect on TN removal in the USFCWs, for the disappearance of some species having anammox ability. Key enzymes of anammox process (NIR and HDH) decreased to some extent under GO and Ag NP exposure, and function gene of defense mechanisms had an increase trend in samples.     

Investigation of physico-chemical properties and microbial community during poultry manure co-composting process

Co-composting of poultry manure and rubber wood sawdust was performed with the ratio of 2:1 (V/V) for a period of 60 days. An investigation was carried out to study the extracellular enzymatic activities and structural degradation utilizing Fourier transform infrared spectroscopy (FT-IR), thermogravimetry and differential thermal analysis (TG/DTA) and scanning electron microscopy (SEM). The microbial succession was also determined by using denaturing gel gradient electrophoresis (DGGE). The compost was able to reach its highest temperature of 71°C at day 3 and stabilized between 30 and 40°C for 8 weeks. CMCase, FPase and β-glucosidase acted synergistically in order to degrade the cellulosic substrate. The xylanase activities increased gradually during the composting and reached the peak value of 11.637 U/g on day 35, followed by a sharp decline. Both LiP and MnP activities reached their peak values on day 35 with 0.431 and 0.132 U/g respectively. The FT-IR spectra revealed an increase in aromaticity and a decrease in aliphatic compounds such as carbohydrates as decomposition proceeded. TGA/DTG data exhibited significant changes in weight loss in compost samples, indicating degradation of organic matter. SEM micrographs showed higher amounts of parenchyma exposed on the surface of rubber wood sawdust at day 60, showing significant degradation. DGGE and 16S rDNA analyses showed that Burkholderia sp., Pandoraea sp., and Pseudomonas sp. were present throughout the composting process. Ornithinibacillus sp. and Castellaniella ginsengisoli were only found in the initial stage of the composting, while different strains of Burkholderia sp. also occurred in the later stage of composting.     

Mixing regime shapes the community assembly process, microbial interaction and proliferation of cyanobacterial species Planktothrix in a stratified lake

Lake mixing influences aquatic chemical properties and microbial community composition,and thus, we hypothesized that it would alter microbial community assembly and interaction. To clarify this issue, we explored the community assembly processes and cooccurrence networks in four seasons at two depths(epilimnion and hypolimnion) in a mesotrophic and stratified lake(Chenghai Lake), which formed stratification in the summer and turnover in the winter. During the stratification period, the epilimni...     

Investigation of physico-chemical properties and microbial community during poultry manure co-composting process

Co-composting of poultry manure and rubber wood sawdust was performed with the ratio of 2:1(V/V) for a period of 60 days. An investigation was carried out to study the extracellular enzymatic activities and structural degradation utilizing Fourier transform infrared spectroscopy(FT-IR), thermogravimetry and differential thermal analysis(TG/DTA)and scanning electron microscopy(SEM). The microbial succession was also determined by using denaturing gel gradient electrophoresis(DGGE). The compost was able to reach its highest temperature of 71°C at day 3 and stabilized between 30 and 40°C for 8 weeks.CMCase, FPase and β-glucosidase acted synergistically in order to degrade the cellulosic substrate. The xylanase activities increased gradually during the composting and reached the peak value of 11.637 U/g on day 35, followed by a sharp decline. Both Li P and Mn P activities reached their peak values on day 35 with 0.431 and 0.132 U/g respectively. The FT-IR spectra revealed an increase in aromaticity and a decrease in aliphatic compounds such as carbohydrates as decomposition proceeded. TGA/DTG data exhibited significant changes in weight loss in compost samples, indicating degradation of organic matter. SEM micrographs showed higher amounts of parenchyma exposed on the surface of rubber wood sawdust at day 60, showing significant degradation. DGGE and 16 S r DNA analyses showed that Burkholderia sp., Pandoraea sp., and Pseudomonas sp. were present throughout the composting process. Ornithinibacillus sp. and Castellaniella ginsengisoli were only found in the initial stage of the composting, while different strains of Burkholderia sp. also occurred in the later stage of composting.