Changes in the structure and function of microbial communities in drinking water treatment bioreactors upon addition of phosphorus

Phosphorus was added as a nutrient to bench-scale and pilot-scale biologically active carbon (BAC) reactors operated for perchlorate and nitrate removal from contaminated groundwater. The two bioreactors responded similarly to phosphorus addition in terms of microbial community function (i.e., reactor performance), while drastically different responses in microbial community structure were detected. Improvement in reactor performance with respect to perchlorate and nitrate removal started within a few days after phosphorus addition for both reactors. Microbial community structures were evaluated using molecular techniques targeting 16S rRNA genes. Clone library results showed that the relative abundance of perchlorate-reducing bacteria (PRB) Dechloromonas and Azospira in the bench-scale reactor increased from 15.2% and 0.6% to 54.2% and 11.7% after phosphorus addition, respectively. Real-time quantitative PCR (qPCR) experiments revealed that these increases started within a few days after phosphorus addition. In contrast, after phosphorus addition, the relative abundance of Dechloromonas in the pilot-scale reactor decreased from 7.1 to 0.6%, while Zoogloea increased from 17.9 to 52.0%. The results of this study demonstrated that similar operating conditions for bench-scale and pilot-scale reactors resulted in similar contaminant removal performances, despite dramatically different responses from microbial communities. These findings suggest that it is important to evaluate the microbial community compositions inside bioreactors used for drinking water treatment, as they determine the microbial composition in the effluent and impact downstream treatment requirements for drinking water production. This information could be particularly relevant to drinking water safety, if pathogens or disinfectant-resistant bacteria are detected in the bioreactors.     

DGGE analysis of 16S rDNA of ammonia-oxidizing bacteria in chemical–biological flocculation and chemical coagulation systems

Microbial community DNA was extracted from activated sludge samples taken from a chemical bioflocculation process and a chemical coagulation process in Shanghai, China. 16S rDNA of ammonia-oxidizing bacteria (AOB)was amplified by nested polymerase chain reaction and fingerprinted by denaturing gradient gel electrophoresis for microbial structure analysis. The Shannon diversity index of each sample was determined. The results indicated that the microbial structure of AOB in chemical bioflocculation process was comparable at two operational conditions. The ammonia-oxidizing bacterial communities were similar in three channels of the chemical bioflocculation process and in three serial tanks in the chemical coagulation process at the same condition. The diversity of microbial structures in the chemical bioflocculation process was higher than in the chemical coagulation process, in which the microbial structure was similar to that in the influent. Although the microbial study provides insights to the nitrification removal, higher microbial diversity of AOB does not necessarily mean higher ammonia oxidization. Molecular analysis should be combined with chemical assays to optimize operational conditions.     

微生物电解池阳极生物膜功能菌群构建及群落特征分析

【目的】微生物电解电池(MEC)是近几年快速发展的利用电极呼吸微生物快速降解有机质,通过较小的辅助外加电压直接生成氢气的新工艺。MEC能够有效地富集高效率电子传递功能菌群,是未来工艺放大和快速启动的关键。【方法】采用不同驯化方法构建MEC电极微生物菌群,通过单链构象多肽性技术(Single-strand conformation poly-morphism,SSCP)快速检测分析启动后电子传递功能菌群特征。【结果】阳极生物膜接种MEC可以实现2 d的快速启动,库仑效率达到20%以上,7 d获得稳定产氢,氢气转化率达到30%,能量回收效率达到90%以上。通过SSCP群落分析发现,采用微生物燃料电池阳极生物膜构建的MEC主要电子传递功能相关的菌群包括Pseudomonas sp.、Flavobacterium sp.、Ochrobactrum sp.,而直接由产氢MEC阳极生物膜新启动的MEC功能菌群组成丰度更大,包括电子传递效能更高的Desulfovibrio、Pseudomonas和Shewanella成为主要优势电子传递菌群。通过稳定产氢运行,MEC阳极生物膜优势菌群中存在的较大比例的厌氧菌与电子传递辅助菌对体系的快速稳定运行十分重要。【结论】与MFC阳极生物膜相比,MEC生物膜作为启动菌源能够获得多样性更丰富的电极功能菌群,其库仑效率和产氢效率更具优势。     

Changes in functional diversity of soil microbial community with addition of antibiotics sulfamethoxazole and chlortetracycline

Potential effects of antibiotics on agricultural soil microflora have recently become increasing concerns with antibiotic-contaminated biosolid now being used in agricultural land. However, changes of soil microbial community function caused by the antibiotic-associated disturbance are less addressed. This paper investigated the changes in microbial functional diversity by spiking sulfamethoxazole (SMX) and chlortetracycline (CTC) in a loam paddy soil and then incubating for 21 days. The dose-effect and time-dependent changes of antibiotic-associated disturbance on soil microbial community were analyzed with the soils sampled at 7 and 21 days using Biolog EcoPlate. At day 7 following treatment, SMX decreased functional diversity of soil microbial community, and the treatment of 100 mg SMX kg?1 dry soil had a significant inhibition of average well color development (AWCD) and Shannon index as compared to the control (p?相似文献   

喹啉与吲哚驯化的反硝化反应器的微生物群落结构分析比较

[目的]本研究旨在比较分析分别以喹啉和吲哚为底物,在相同条件下驯化的两个反硝化生物反应器的微生物群落结构.[方法]采用相同的种子污泥和相同的驯化条件,经过大约6周的驯化后,两个反应器均达到稳定而高效的污染物去除能力,通过16S rDNA克隆文库技术对两个反应器的微生物群落结构进行研究.[结果]研究发现,微生物群落结构表现出很大的差异.喹啉驯化的群落中所有的OTU都属于Betaproteobacteria,而吲哚驯化的群落中Betaproteobacteria占56.3%,吲哚驯化的群落具有更高的多样性.两个群落的优势OTU也不同,喹啉驯化群落中Thauera及其它Rhodocyclaceae科的微生物占整个群落的73%,而吲哚驯化群落中优势OTU为Comamonadaceae科、Alcaligenaceae科和Rhodocyclaceae科等类型的微生物,其中Comamonadaceae科的一个OTU占整个群落的28.7%.[结论]不同的驯化底物对微生物群落的组成具有较强的选择作用.首次报道并比较了可高效降解喹啉和吲哚的反硝化生物反应器的微生物群落结构.     

废气处理生物滤器微生物生态学研究进展

废气生物处理是一项新兴的气体污染控制技术,已成为当前环境领域的研究热点.本文概述了生物滤器系统(包括生物过滤器系统和生物滴滤器系统)结构及其去除气体污染物的原理,重点阐述了生物滤器微生物的分离、鉴定及其降解特性,微生物丰度、活性与微生物群落结构多样性之间的相关性,运行条件对微生物群落的影响,微生物群落结构时空变化规律,生物膜形成机理和模型等方面的研究进展,并对今后废气处理生物滤器微生物生态学研究方向进行了展望.     

Spatial and temporal changes in microbial community structure associated with recharge-influenced chemical gradients in a contaminated aquifer

In a contaminated water-table aquifer, we related microbial community structure on aquifer sediments to gradients in 24 geochemical and contaminant variables at five depths, under three recharge conditions. Community amplified ribsosomal DNA restriction analysis (ARDRA) using universal 16S rDNA primers and denaturing gradient gel electrophoresis (DGGE) using bacterial 16S rDNA primers indicated: (i). communities in the anoxic, contaminated central zone were similar regardless of recharge; (ii). after recharge, communities at greatest depth were similar to those in uncontaminated zones; and (iii). after extended lack of recharge, communities at upper and lower aquifer margins differed from communities at the same depths on other dates. General aquifer geochemistry was as important as contaminant or terminal electron accepting process (TEAP) chemistry in discriminant analysis of community groups. The Shannon index of diversity (H) and the evenness index (E), based on DGGE operational taxonomic units (OTUs), were statistically different across community groups and aquifer depths. Archaea or sulphate-reducing bacteria 16S rRNA abundance was not clearly correlated with TEAP chemistry indicative of methanogenesis or sulphate reduction. Eukarya rRNA abundance varied by depth and date from 0 to 13% of the microbial community. This contaminated aquifer is a dynamic ecosystem, with complex interactions between physical, chemical and biotic components, which should be considered in the interpretation of aquifer geochemistry and in the development of conceptual or predictive models for natural attenuation or remediation.     

Relationship between communities and processes; new insights from a field study of a contaminated ecosystem

We used a 93‐year‐old mine waste contamination gradient in alluvial soil to explore the relationship between ecosystem level functioning and community structure in a chronically stressed ecosystem. The sensitivity of broad functional parameters (in situ soil respiration, microbial biomass, above and below ground plant biomass) and microbial diversity [phospholipid fatty acid (PLFA) abundance and richness] were compared. Functional responses were linear with respect to contaminants while thresholds were detected in the community structural response to contamination along the gradient. For example, in situ soil respiration was negatively and linearly correlated to contamination concentration (R = ?0.783, P < 0.01), but changes in microbial community structure only became evident where contaminant concentrations were greater than 28 times above background levels. Our results suggest that functional redundancy does not prevent depression of ecosystem function in the long‐term.     

生物膜法强化净化氨氮污染水体及其微生物群落解析

【目的】比较不同营养条件及挂膜方式下生物膜法对氨氮污染水体的净化效果及其功能微生物群落结构。【方法】设置空白(Blank)、自然成膜(Raw)、预附脱氮菌强化挂膜(PCC)3组生物膜反应器,利用末端限制性片段长度多态性(T-RFLP)技术和非度量多维标度(NMDS)分析方法对生物膜反应器转化氨氮过程中微生物群落结构及其演替过程进行动态解析。【结果】在C/N=1:1时,除PCC在起始阶段短暂具有较高的氨氮脱除效率外,Blank、Raw和PCC最终均表现出较低的氨氮转化效率(10%-20%)。改变C/N=2:1后,Raw和PCC对人工合成污水中NH4+-N的转化率均提高至95%以上,而且Raw与PCC的群落结构在C/N=2:1时具有较高的相似性,优势菌群主要为γ-变形菌纲(Gammaproteobacteria)、放线菌纲(Actinobacteria)和硝化螺菌纲(Nitrospira)。【结论】C/N是影响生物膜反应器氨氮去除效果及驱动生物膜反应器中细菌群落结构发生改变的重要因子。     

GeoChip-based analysis of functional microbial communities during the reoxidation of a bioreduced uranium-contaminated aquifer

A pilot-scale system was established for in situ biostimulation of U(VI) reduction by ethanol addition at the US Department of Energy's (DOE's) Field Research Center (Oak Ridge, TN). After achieving U(VI) reduction, stability of the bioreduced U(IV) was evaluated under conditions of (i) resting (no ethanol injection), (ii) reoxidation by introducing dissolved oxygen (DO), and (iii) reinjection of ethanol. GeoChip, a functional gene array with probes for N, S and C cycling, metal resistance and contaminant degradation genes, was used for monitoring groundwater microbial communities. High diversity of all major functional groups was observed during all experimental phases. The microbial community was extremely responsive to ethanol, showing a substantial change in community structure with increased gene number and diversity after ethanol injections resumed. While gene numbers showed considerable variations, the relative abundance (i.e. percentage of each gene category) of most gene groups changed little. During the reoxidation period, U(VI) increased, suggesting reoxidation of reduced U(IV). However, when introduction of DO was stopped, U(VI) reduction resumed and returned to pre-reoxidation levels. These findings suggest that the community in this system can be stimulated and that the ability to reduce U(VI) can be maintained by the addition of electron donors. This biostimulation approach may potentially offer an effective means for the bioremediation of U(VI)-contaminated sites.     

Operational and microbiological aspects of a bioaugmented two-stage biotrickling filter removing hydrogen sulfide and dimethyl sulfide

A two-stage biotrickling filter was developed for removing dimethyl sulfide (DMS) and hydrogen sulfide (H2S). The first biotrickling filter (ABF) was inoculated with Acidithiobacillus thiooxidans and operated without pH control, while the second biotrickling filter (HBF) was inoculated with Hyphomicrobium VS and operated at neutral pH. High DMS elimination capacities were observed in the HBF (8.2 g DMS m(-3) h(-1) at 90% removal efficiency) after 2 days. Maximal observed elimination capacities were 83 g H2S m(-3) h(-1) (100% removal efficiency) and 58 g DMS m(-3) h(-1) (88% removal efficiency) for the ABF and the HBF, respectively. The influence of a decreasing empty bed residence time (120 down to 30 sec) and the robustness of the HBF towards changing operational parameters (low pH, starvation, and DMS and H2S peak loadings) were investigated. Suboptimal operational conditions rapidly resulted in lower DMS removal efficiencies, but recovery of the HBF was mostly obtained within a few days. The H2S removal efficiency in the ABF, however, was not influenced by varying operational conditions. In both reactors, microbial community dynamics of the biofilm and the suspended bacteria were investigated, using denaturing gradient gel electrophoresis (DGGE). After a period of gradual change, a stable microbial community was observed in the HBF after 60 days, although Hyphomicrobium VS was not the dominant microorganism. In contrast, the ABF biofilm community was stable from the first day and only a limited bacterial diversity was observed. The planktonic microbial community in the HBF was very different from that in the biofilm.     

Bacterial diversity and bioaugmentation in floodwater of a paddy field in the presence of the herbicide molinate

This work aimed at studying variations on the diversity and composition of the bacterial community of a rice paddy field floodwater, subjected to conventional management, namely by using the herbicide molinate. The promotion of the herbicide biodegradation either by the autochthonous microbiota or by a bioaugmentation process was also assessed. This study comprehended four sampling campaigns at key dates of the farming procedures (seeding, immediately and 6 days after application of the herbicide molinate, and after synthetic fertilization) and the subsequent physic-chemical and microbiological characterization (pH, DOC and molinate contents, total cells, cultivable bacteria and DGGE profiling) of the samples. Multivariate analysis of the DGGE profiles showed temporal variations in the bacterial community structure and the Shannon’s index values indicated that the bacterial diversity reached its minimum at the molinate application day. The highest bacterial diversity coincided with the periods with undetectable concentrations of the herbicide, although microcosm assays suggested that other factors than molinate may have been responsible for the decrease of the bacterial diversity. The ability of autochthonous microorganisms to degrade molinate and the influence of the herbicide on the bacterial community composition were assessed in microcosm assays using floodwater collected at the same dates. Given molinate was not degraded by autochthonous microorganisms, and considering it represents an environmental contaminant, bioaugmentation microcosms were assayed aiming the assessment of the feasibility of a bioremediation process to clean contaminated floodwater. A molinate-mineralizing culture, previously isolated, promoted molinate removal, induced alterations in the autochthonous bacterial community structure and diversity, and was undetected after 7 days of incubation, suggesting the feasibility of the process.     

DG-DGGE分析产氢发酵系统微生物群落动态及种群多样性

应用双梯度-变性梯度凝胶电泳(DG-DGGE)对生物制氢反应器微生物种群的动态变化及多样性进行监测。间隔7d从反应器取厌氧活性污泥,以细菌16SrDNA通用引物进行V2～V3区域PCR扩增,长约450bp的PCR产物经DGGE分离后,获得污泥微生物群落的16SrDNA指纹图谱。污泥接种到反应器后微生物群落中既有原始种群的消亡和增长,也有次级种群的强化和演变。反应器在运行初期群落演替迅速,15d时微生物群落结构变化最大。群落结构的相似性随着演替时间的增加而逐渐升高,种群动态变化后形成稳定的群落结构。29d时微生物多样性基本保持不变,微生物优势种属达到19个OTU。在细菌竞争和协同作用制约下,种群多样性降低后趋于稳定,形成顶级群落。有些种群在群落结构中一直存在,是群落建成的原始种群,原始种群与次级种群在代谢过程中具有协同作用,表现出群落的综合生态特征。     

Degradation of Polycyclic Aromatic Hydrocarbons in the Rhizosphere of Festuca arundinacea and Associated Microbial Community Changes

A phytoremediation growth chamber study was conducted to evaluate the contribution of soil microbial diversity to the contaminant degradation. Target contaminant removal from soil was assessed by monitoring concentrations of polycyclic aromatic hydrocarbons (PAHs), along with changes in the bacterial community structure over a time period of 10 months in the presence of tall fescue (Festuca arundinacea). Enhanced degradation of PAHs was observed in rhizosphere soil, with a maximum reduction in pyrene at a rate 36% higher than that noted for the unvegetated control. The dissipation of < 4-ring PAHs, 4-ring PAHs, and > 4-ring PAHs in unvegetated soil was 70%, 54%, and 49% respectively, whereas a higher dissipation rate was observed in tall fescue treated soil of 78%, 68%, and 61% at the end of the study. Microbial enumeration results showed greater total bacterial numbers and PAH-degrading bacteria in rhizosphere soil when compared to unvegetated soil. The results from the terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that there was a shift in the rhizosphere bacterial community during the phytoremediation process.     

土壤微生物群落对麻栎-刺槐混交林凋落物分解的影响

以麻栎-刺槐混交林和麻栎纯林为研究对象,采用野外定点采样、室内分析与高通量测序的方法,对凋落物分解过程中土壤微生物菌群多样性特征及其对凋落物分解速率的影响进行了研究。结果表明:(1)麻栎-刺槐混交林凋落物的分解速率高于麻栎纯林。两种林分凋落物有机碳(TOC)、全氮(TN)发生释放,全磷(TP)发生积累-释放的过程。(2)两种林分土壤细菌优势类群为放线菌门(Acidobacteria)、变形菌门(Proteobacteria)、酸杆菌门(Actinobacteria)和疣微菌门(Verrucomicrobia),土壤真菌优势类群为担子菌门(Basidiomycota)、子囊菌门(Ascomycota)和被孢霉门(Moritierellomycota)。(3)凋落物分解过程中,麻栎-刺槐混交林土壤微生物菌群丰富度指数和菌群多样性指数变化范围小于麻栎纯林。(4)凋落物分解速率与土壤细菌菌群丰富度指数和菌群多样性指数呈显著正相关,与土壤真菌菌群丰富度指数呈显著正相关。土壤微生物群落对麻栎-刺槐混交林和麻栎纯林凋落物分解速率具有重要影响,研究结果为深入开展混交林土壤微生物多样性对凋落物分解的影响研究提供理论依据。     

Long-term effect of re-vegetation on the microbial community of a severely eroded soil in sub-tropical China

The long-term (18 years) effects of re-vegetating eroded soil on soil microbial biomass, community structure and diversity were investigated in a forest soil derived from Quaternary clay in the Red Soil Ecological Experimental Station of the Chinese Academy of Sciences. Large areas of land in this region of China have been subjected to severe soil erosion, characterised by the removal of the fertile surface soil and even the exposure of parental rock in some areas due to a combination of deforestation and heavy rainfall. The effects of planting eroded or uneroded soil with Pinus massoniana, Cinnamomum camphora or Lespedeza bicolor on the soil microbial community and chemical properties were assessed. Total soil microbial community DNA was extracted and bacterial 16 S rRNA gene fragments were amplified by PCR and analysed by terminal restriction fragment length polymorphism (T-RFLP). Microbial biomass carbon (C_mic) was measured by chloroform fumigation-extraction. Following the restoration there were significant increases in both C_mic and bacterial diversity (Shannon index), and significant changes in bacterial community structure. Erosion factors were significant only in minor dimensions suggesting that the restoration had been largely successful in terms of bacterial community structure. Compared with uneroded soil, C_mic recovered in L. bicolor and P. massoniana restored eroded plots and was significantly greater under these tree species than C. camphora, although soils in C. camphora restored plots displayed the highest bacterial diversity. The recovery of microbial biomass and diversity in the eroded plots was, to large extent, accompanied by the development of the same bacterial community structure as in the uneroded plots with erosion having relatively little effect on bacterial community structure.     

Differences in vegetation composition and plant species identity lead to only minor changes in soil-borne microbial communities in a former arable field

To examine the relationship between plant species composition and microbial community diversity and structure, we carried out a molecular analysis of microbial community structure and diversity in two field experiments. In the first experiment, we examined bacterial community structure in bulk and rhizosphere soils in fields exposed to different plant diversity treatments, via a 16S rRNA gene clone library approach. Clear differences were observed between bacterial communities of the bulk soil and the rhizosphere, with the latter containing lower bacterial diversity. The second experiment focused on the influence of 12 different native grassland plant species on bacterial community size and structure in the rhizosphere, as well as the structure of Acidobacteria and Verrucomicrobia community structures. In general, bacterial and phylum-specific quantitative PCR and PCR-denaturing gradient gel electrophoresis revealed only weak influences of plant species on rhizosphere communities. Thus, although plants did exert an influence on microbial species composition and diversity, these interactions were not specific and selective enough to lead to major impacts of vegetation composition and plant species on below-ground microbial communities.     

造纸废水活性污泥的微生物群落结构及多样性分析

为探究造纸废水活性污泥中微生物群落结构多样性以及对造纸废水处理效果的影响，利用Illumina MiSeq 高通量测序方法，分析在处理造纸废水过程中，同一运行阶段两个并联氧化沟内活性污泥的微生物群落与多样性组成。结果表明，系统中处理造纸废水的活性污泥在同一废水条件下微生物群落结构总体稳定，优势细菌为绿弯菌门(Chloroflexi)、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）、Myxococcota、放线菌门(Actinobacteria）、厚壁菌门(Firmicutes）等。最重要的优势细菌类群为Chloroflexi，相对丰度占比为47.67%~48.22%，远远高于其他废水中Chloroflexi的占比，其中厌氧绳菌纲（Anaerolineae）是其主要成员，占比84.39%~88.34%，可针对性地去除造纸废水中的污染物。造纸废水活性污泥样品中存在大量特殊功能菌群，其在废水中污染物尤其是木质素的去除中发挥着重要作用。     

长期人为干扰对桂西北喀斯特草地土壤微生物多样性及群落结构的影响

人为干扰是喀斯特草地正向演替的主要障碍因素, 从土壤微生物角度探讨草地对干扰的响应可为喀斯特生态恢复与科学利用提供依据。该研究依托中国科学院环江喀斯特生态系统观测站长期干扰监测样地, 通过高通量测序测定3个干扰模式(火烧、刈割、刈割除根)和1个对照(封育)处理的土壤细菌、真菌α多样性和群落结构, 并分析它们与环境因子(坡位、土壤理化性质)的关系, 揭示人为干扰对桂西北喀斯特草地土壤微生物的影响及机制。结果显示: 1)对于α多样性, 在中坡和下坡, 火烧均显著降低了真菌Chao1指数, 刈割均显著降低了细菌的香农指数和谱系多样性指数; 刈割除根在中、下坡分别显著降低了真菌Chao1指数和细菌香农指数。2)对于群落结构, 火烧、刈割和刈割除根在中、下坡均显著降低了酸杆菌门细菌相对丰度, 火烧在下坡使子囊菌门真菌相对丰度从74.49%显著降低至34.72%。3)冗余分析显示, 微生物生物量碳含量解释细菌和真菌α多样性变化的29.8%和26.8%以及细菌群落结构变化的31.7%; 细根生物量解释细菌α多样性和真菌群落结构变化的13.9%和10.3%。可见, 长期人为干扰对土壤微生物α多样性产生负面影响, 并显著改变微生物群落结构, 影响程度因干扰方式、微生物种类不同而异, 且受坡位调控。土壤微生物生物量碳含量和细根生物量的减少是人为干扰影响土壤微生物多样性和群落结构的重要因素。α多样性及子囊菌的减少将不利于土壤生态系统稳定性的维持, 酸杆菌的减少将不利于土壤有机质降解和铁循环。因此, 长期火烧、刈割等人为干扰将导致喀斯特草地生态系统功能发生退化。