污水脱氮功能微生物的组学研究进展

生物脱氮是污水处理厂的核心,掌握生物脱氮过程相关微生物代谢特性,对于探索微生物资源和提高污水处理厂脱氮性能具有重要意义。近年来,分子生物学方法不断发展和改进,已被广泛应用于揭示脱氮微生物群落多样性、组成结构和潜在功能等方面,大幅提升了研究者们对污水生物脱氮系统中微生物,尤其是不可培养微生物的代谢机理、抑制调控原理及新型生物脱氮工艺途径的认识。本文对流行的分子生物学方法(16S rRNA基因测序、实时荧光定量PCR技术、宏基因组学、宏转录组学、宏蛋白质组学和代谢组学)进行了介绍,综述了其在硝化细菌、反硝化细菌、完全氨氧化细菌、厌氧氨氧化细菌、厌氧铁氨氧化细菌、硫酸盐型厌氧氨氧化细菌及亚硝酸盐/硝酸盐型厌氧甲烷氧化微生物等方面的研究进展,阐明了这些氮素转化微生物在氮循环过程的代谢途径和酶促反应,并从标准测定方法构建、不同方法的联用及跨学科结合和检测方法的简易化这3个方面展望了分子生物学方法的技术突破及其在污水生物处理系统中的应用前景。本综述从系统角度全面认识脱氮微生物群落及其结构,为未来污水处理生物脱氮微生物的研究提供了新方向。     

固相反硝化系统中微生物群落结构的研究进展

固相反硝化是一种新型的异养反硝化工艺,采用固体有机物同时作为反硝化微生物的碳源和生物膜载体,可用于地下水和低C/N比污水的脱氮处理。固相反硝化系统生物膜的微生物群落结构决定固体碳源的降解效率,进而决定反硝化脱氮的速率和系统的稳定运行。因此,微生物群落结构的研究对于固相反硝化工艺的优化以及反应机理的解析具有重要意义。对不同固相反硝化系统微生物群落结构的研究现状和进展进行了综述,并探讨了当前研究中存在的问题和发展趋势。     

人工湿地氮去除关键功能微生物生态学研究进展

人工湿地是一种能有效处理水体氮素污染的生态技术,其中微生物是驱动人工湿地系统中氮素去除的重要引擎。近20年来,随着分子生物学技术的广泛应用,有关人工湿地氮去除功能微生物生态学方面研究取得了一些重要进展。以硝化-反硝化作用和厌氧氨氧化作用这两种重要的人工湿地微生物脱氮途径为主,针对氨氧化细菌/古菌、厌氧氨氧化菌和反硝化菌等关键脱氮功能微生物的研究,重点归纳总结了目前有关这几类关键功能菌群在人工湿地中的丰度、活性、多样性、分布特征与影响因素,及其对废水中氮去除的作用,并在此基础上对今后的重点研究工作提出了展望。面向未来人工湿地氮去除关键功能微生物的研究应侧重其在污水净化和温室气体减排等方面的生态功能研究,同时加强其代谢过程与机制以及不同功能菌群间的关联研究。     

微生物生物膜研究的新进展

微生物在生长过程中为适应生存环境而形成了生物膜,Dr.Costerton JW在生物膜方面的研究为我们开拓了微生物学的新领域。微生物生物膜是由微生物群体及其包被的细胞外多聚物和基质网组成,它们彼此黏附或者黏附到组织或物体的表面。微生物生物膜与微生物的耐药性形成、基因的转移以及引起机体的持续性感染等都密切相关。目前对生物膜的研究重点已经深入到微生物相互间的信号传递、致病基因的转移以及如何干预微生物生物膜的形成等方面。此外,在治理污水和环境保护工程、生物材料工程和食品工业等方面,微生物生物膜技术已经得到了应用。     

全程硝化菌微生物学特性及在水处理领域的应用潜力

全程硝化菌是近期微生物氮循环领域的重大发现之一,引发了对其全球分布、系统发育特征和生理生化特性的广泛关注。本文综述了全程硝化菌在土壤、地表水、废水处理系统等生境的分布规律及影响因子;并从底物亲和力、代谢多样性等方面阐述了其与传统硝化微生物间的竞争互作和生态位分离机制;基于上述特征提出全程硝化菌在水处理领域中的应用前景,可能与其他脱氮微生物如反硝化菌、厌氧氨氧化菌和厌氧甲烷氧化菌等耦合实现在低氨氮、低溶解氧条件下的污水深度脱氮,从而节省能耗并降低温室气体排放。未来研究应继续深入研究全程硝化菌的生理生化特性,评价其生态功能和对氮素地球化学循环的贡献,并探索其在生物水处理等领域的应用潜力。     

聚磷生物膜法磷回收研究进展

从污水中回收磷已经成为当前污水全面资源化研究领域的重要研究方向之一,聚磷生物膜法磷回收工艺因其在节省碳源、简化回收流程、提高回收效果、污泥减量化等方面的潜力,成为该领域的重要技术手段和研究热点。本文着重介绍了聚磷生物膜法磷回收的基本原理、功能微生物及其代谢特征;详细梳理了生物膜法磷回收工艺不同模式的回收思路,综合分析了不同工艺模式的技术特征和磷回收效果。此外,对聚磷生物膜系统微生物及其代谢的研究方法与技术进行了整理。文章在综合分析聚磷生物膜法磷回收研究现状的基础上,对该技术的应用前景进行了宏观展望,以期为开展聚磷生物膜法磷回收的基础研究和技术开发提供支撑。     

反硝化细菌在污水脱氮中的作用

反硝化是在反硝化细菌的作用下,以硝酸盐作为最终电子受体而进行的无氧呼吸过程。从污水脱氮的角度论述反硝化在污水脱氮中的作用、污水脱氮的机理、污水脱氮过程中反硝化作用的影响因素等。从反硝化的角度出发,论述了反硝化细菌的类群、反硝化作用的机理、反硝化细菌细胞中参与反硝化过程的关键酶。另外,还论述了近年来发现的有氧反硝化细菌、自养反硝化细菌及反硝化除磷细菌等方面的研究进展。     

生物膜中不同种属微生物的交流与合作

自然界中的微生物可附着于载体表面形成高度组织化、系统化的微菌落膜性聚合物, 即生物膜. 与浮游状态的微生物不同, 生物膜中不同种属微生物进行着复杂的交流与合作, 产生“1+1>2”的非线性相互作用, 从而使生物膜在整体上表现出一系列新的生物学特征. 生物膜给人们的生活生产带来很多不利的影响, 如引起顽固性感染性疾病威胁人类健康、破坏输水管道系统导致工业损失等; 但另一方面, 生物膜也可发挥积极作用, 如降解污染物修复环境, 形成生物屏障保护生态等. 如何认识生物膜、如何趋利避害使生物膜造福于人类已成为当今世界多领域关注和研究的重要课题. 从生物膜的形成、代谢产物与信号分子、水平基因转移及其与外界环境的关系等方面对生物膜中交流与合作的研究进展及意义进行了综述, 并提出应从“微生物组学”的高度整体认识生物膜.     

影响变形链球菌生物膜形成因素的研究进展

龋病的发生发展过程中,致龋微生物生物膜(biofilm)形成是龋病发生的首要因素。变形链球菌(Streptococcus mutans)等在口腔生物膜内进行产酸代谢活动是产生龋病的直接原因。因此,控制S.mutans生物膜的形成可以减少龋病的发生、发展。本文就影响变形链球菌生物膜形成的因素研究进展做一综述。     

土壤氮素转化的关键微生物过程及机制

微生物是驱动土壤元素生物地球化学循环的引擎.氮循环是土壤生态系统元素循环的核心之一,其四个主要过程,即生物固氮作用、氨化作用、硝化作用、反硝化作用,均由微生物所驱动.近10年来,随着免培养的分子生态学技术和高通量测序技术等的发展,在硝化微生物多样性及其作用机理、厌氧氨氧化过程和机理等研究方面取得了突破性进展.本文重点阐述了我国有关土壤硝化微生物方面的研究进展,在此基础上,简要介绍了反硝化微生物和厌氧氨氧化及硝酸盐异化还原成铵作用的研究进展,并对今后的研究工作提出了展望.今后土壤氮素转化微生物生态学的研究,应瞄准国际微生生态学发展的前沿,加强新技术新方法的应用,结合我国农业可持续发展、资源环境保护和全球变化研究的重大需求,重点开展以下几方面的工作:(1)开展大尺度上土壤硝化作用及氨氧化微生物分布的时空演变特征及驱动因子的研究;(2)加强氮素转化关键微生物过程与机理的研究,并与相关过程的通量(如氨挥发、N2O释放)和反应速率(如矿化速率、硝化速率)关联起来;(3)在特定生态系统中系统研究各个氮转化过程的耦合关系,构建相关氮素转化和氮素平衡模型,为定向调控土壤氮素转化过程,提高氮素利用效率并减少其负面效应提供科学依据.     

Performance and Microbial Structure of a Combined Biofilm Reactor

A novel combined biofilm reactor was established and applied as a single treatment unit for carbon and nitrogen removal of wastewater. The nitrogen removal performance of the reactor at different levels of organic carbon (COD) loading was investigated when the influent total nitrogen (TN) loading was 0.74 g TN/m² day. Continuous experimental results demonstrated that 80% nitrogen was eliminated when the influent COD loading ranged between 2.06 g and 3.92 g COD/m² day. Microbial composition in the reactor was analyzed using fluorescent in situ hybridization (FISH) and conventional batch tests. The relative abundance of ammonia-oxidizing bacteria in the aerobic zone of the reactor measured by FISH was consistent with the result from conventional batch tests.     

Sensitivity analysis of a biofilm model describing a one-stage completely autotrophic nitrogen removal (CANON) process.

A mathematical model for nitrification and anaerobic ammonium oxidation (ANAMMOX) processes in a single biofilm reactor (CANON) was developed. This model describes completely autotrophic conversion of ammonium to dinitrogen gas. Aerobic ammonium and nitrite oxidation were modeled together with ANAMMOX. The sensitivity of kinetic constants and biofilm and process parameters to the process performance was evaluated, and the total effluent concentrations were, in general, found to be insensitive to affinity constants. Increasing the amount of biomass by either increasing biofilm thickness and density or decreasing porosity had no significant influence on the total effluent concentrations, provided that a minimum total biomass was present in the reactor. The ANAMMOX process always occurred in the depth of the biofilm provided that the oxygen concentration was limiting. The optimal dissolved oxygen concentration level at which the maximum nitrogen removal occurred is related to a certain ammonium surface load on the biofilm. An ammonium surface load of 2 g N/m2. d, associated with a dissolved oxygen concentration level of 1.3 g O2/m3 in the bulk liquid and with a minimum biofilm depth of 1 mm seems a proper design condition for the one-stage ammonium removal process. Under this condition, the ammonium removal efficiency is 94% (82% for the total nitrogen removal efficiency) (30 degrees C). Better ammonium removal could be achieved with an increase in the dissolved oxygen concentration level, but this would strongly limit the ANAMMOX process and decrease total nitrogen removal. It can be concluded that a one-stage process is probably not optimal if a good nitrogen effluent is required. A two-stage process like the combined SHARON and ANAMMOX process would be advised for complete nitrogen removal.     

Rotating algal biofilm reactor and spool harvester for wastewater treatment with biofuels by-products

Maximizing algae production in a wastewater treatment process can aid in the reduction of soluble nitrogen and phosphorus concentrations in the wastewater. If harvested, the algae-based biomass offers the added benefit as feedstock for the production of biofuels and bioproducts. However, difficulties in harvesting, concentrating, and dewatering the algae-based biomass have limited the development of an economically feasible treatment and production process. When algae-based biomass is grown as a surface attached biofilm as opposed to a suspended culture, the biomass is naturally concentrated and more easily harvested. This can lead to less expensive removal of the biomass from wastewater, and less expensive downstream processing in the production of biofuels and bioproducts. In this study, a novel rotating algal biofilm reactor (RABR) was designed, built, and tested at bench (8 L), medium (535 L), and pilot (8,000 L) scales. The RABR was designed to operate in the photoautotrophic conditions of open tertiary wastewater treatment, producing mixed culture biofilms made up of algae and bacteria. Growth substrata were evaluated for attachment and biofilm formation, and an effective substratum was discovered. The RABR achieved effective nutrient reduction, with average removal rates of 2.1 and 14.1 g m(-2) day(-1) for total dissolved phosphorus and total dissolved nitrogen, respectively. Biomass production ranged from 5.5 g m(-2) day(-1) at bench scale to as high as 31 g m(-2) day(-1) at pilot scale. An efficient spool harvesting technique was also developed at bench and medium scales to obtain a concentrated product (12-16% solids) suitable for further processing in the production of biofuels and bioproducts.     

Redox-stratification controlled biofilm (ReSCoBi) for completely autotrophic nitrogen removal: the effect of co- versus counter-diffusion on reactor performance

A multi-population biofilm model for completely autotrophic nitrogen removal was developed and implemented in the simulation program AQUASIM to corroborate the concept of a redox-stratification controlled biofilm (ReSCoBi). The model considers both counter- and co-diffusion biofilm geometries. In the counter-diffusion biofilm, oxygen is supplied through a gas-permeable membrane that supports the biofilm while ammonia (NH(4)(+)) is supplied from the bulk liquid. On the contrary, in the co-diffusion biofilm, both oxygen and NH(4)(+) are supplied from the bulk liquid. Results of the model revealed a clear stratification of microbial activities in both of the biofilms, the resulting chemical profiles, and the obvious effect of the relative surface loadings of oxygen and NH(4)(+) (J(O(2))/J(NH(4)(+))) on the reactor performances. Steady-state biofilm thickness had a significant but different effect on T-N removal for co- and counter-diffusion biofilms: the removal efficiency in the counter-diffusion biofilm geometry was superior to that in the co-diffusion counterpart, within the range of 450-1,400 microm; however, the efficiency deteriorated with a further increase in biofilm thickness, probably because of diffusion limitation of NH(4)(+). Under conditions of oxygen excess (J(O(2))/J(NH(4)(+)) > 3.98), almost all NH(4)(+) was consumed by aerobic ammonia oxidation in the co-diffusion biofilm, leading to poor performance, while in the counter-diffusion biofilm, T-N removal efficiency was maintained because of the physical location of anaerobic ammonium oxidizers near the bulk liquid. These results clearly reveal that counter-diffusion biofilms have a wider application range for autotrophic T-N removal than co-diffusion biofilms.     

Performance of a down-flow fluidized bed reactor under sulfate reduction conditions using volatile fatty acids as electron donors

The use of a down-flow fluidized bed (DFFB) reactor for the treatment of a sulfate-rich synthetic wastewater was investigated to obtain insight into the outcome of sulfate reduction in a biofilm attached to a plastic support under a down-flow regime. Fine low-density polyethylene particles were used as support for developing a biofilm within the reactor. The reactor treated a volatile fatty acids mixture of acetate or lactate, propionate, and butyrate at different chemical oxygen demand (COD) to sulfate ratios ranging from 1.67 to 0.67 (g/g). Organic loading rate changed from 2.5 to 5 g COD/L x day and sulfate loading rate increased from 1.5 to 7.3 g SO(4) (2-)/L x day. At the beginning of continuous operation, methanogenesis was the predominant process; however, after 187 days, sulfate reduction became the main ongoing biological process. After 369 days, a COD removal of 93% and a sulfate removal of 75% were reached. Total sulfide concentrations in the reactor ranged from 105, when the reactor was mainly methanogenic, to around 1,215 mg/L at the end of the experiment. The high sulfide concentrations did not affect the performance of the reactor. Results demonstrated that the configuration of the DFFB reactor was suitable for the anaerobic treatment of sulfate-rich wastewater.     

Sequencing batch membrane biofilm reactor for simultaneous nitrogen and phosphorus removal: novel application of membrane-aerated biofilm

A sequencing batch membrane biofilm reactor (SBMBfR) was developed for simultaneous carbon, nitrogen, and phosphorus removal from wastewater. This reactor was composed of two functional parts: (1) a gas-permeable membrane on which a nitrifying biofilm formed and (2) a bulk solution in which bacteria, mainly denitrifying polyphosphate-accumulating organisms (DNPAOs), were suspended. The reactor was operated sequentially under anaerobic condition and then under membrane aeration condition in one cycle. During the anaerobic period, organic carbon was consumed by DNPAOs; this was accompanied by phosphate release. During the subsequent membrane aeration period, nitrifying bacteria utilized oxygen supplied directly to them from the inside of the membrane. Consequently, the nitrite and nitrate products diffused into the bulk solution, where they were used by DNPAOs as electron acceptors for phosphate uptake. In a long-term sequencing batch operation, the mean removal efficiencies of total organic carbon (TOC), total nitrogen (T-N), and total phosphorus (T-P) under steady-state condition were 99%, 96%, and 90%, respectively. In addition, fluorescence in situ hybridization (FISH) clearly demonstrated the difference in bacterial community structure between the membrane biofilm and the suspended sludge: ammonia-oxidizing bacteria belonging to the Nitrosomonas group were dominant in the region adjacent to the membrane throughout the operation, and the occupation ratio of the well-known polyphosphate-accumulating organism (PAO) Candidatus "Accumulibacter phosphates" in the suspended sludge gradually increased to a maximum of 37%.     

Anaerobic treatment of highly concentrated aniline wastewater using packed-bed biofilm reactor

The performance of packed-bed biofilm reactor (PBBR) with self-floating bio-carriers was investigated to treat highly concentrated organic nitrogenous aniline wastewater with a COD value as high as 24,000 mg/L. With 45 vol% of carrier charge inside the reactor, the aniline wastewater can be effectively treated with 94% of COD removal efficiency at a low organic loading rate (OLR) of 0.9 kg COD/(m³ d). The removal efficiency decreased gradually down to 75% when OLR increased to 12.27 kg COD/(m³ d) that corresponded to 1 day of HRT. Separate tests with biofilm alone showed that the conversion contribution of the biofilm was about half of the overall COD conversion by the biofilm plus sludge system at the same OLRs of 3–4 kg COD/(m³ d), and that the biofilm had higher activity than suspended sludge. Ammonium released from decomposed aniline was increased gradually from 500 to 1700 mg/L with the OLR increase from 0.9 to 12.27 kg COD/(m³ d), which resulted in inhibitory effect to the microorganism due to the toxicity of free ammonia. Batch anaerobic toxicity tests showed that the biofilm was less sensitive to toxic compounds than suspended sludge and could tolerate higher concentration of free ammonia.     

Rapid startup and high rate nitrogen removal from anaerobic sludge digester liquor using a SNAP process

In this study, a single-stage autotrophic nitrogen removal reactor, packed with a novel acrylic fiber biomass carrier material (Biofix), was applied for nitrogen removal from sludge digester liquor. For rapid start-up, conventional activated sludge was added to the reactor soon after the attachment of anammox biomass on the Biofix carriers, which allowed conventional activated sludge to form a protective layer of biofilm around the anammox biomass. The Nitrogen removal efficiency reached 75% within 1 week at a nitrogen loading rate of 0.46 kg-N/m³/day for synthetic wastewater treatment. By the end of the synthetic wastewater treatment period, the maximum nitrogen removal rate had increased to 0.92 kg-N/m³/day at a nitrogen loading rate of 1.0 kg-N/m³/day. High nitrogen removal rate was also achieved during the actual raw digester liquor treatment with the highest nitrogen removal rate being 0.83 kg-N/m³/day at a nitrogen loading rate of 0.93 kg-N/m³/day. The thick biofilm on Biofix carriers allowed anammox bacteria to survive under high DO concentration of 5–6 mg/l resulting in stable and high nitrogen removal performance. FISH and CLSM analysis demonstrated that anammox bacteria coexisted and surrounded by ammonium oxidizing bacteria.     

Importance of the operating pH in maintaining the stability of anoxic ammonium oxidation (anammox) activity in moving bed biofilm reactors

Two bench-scale parallel moving bed biofilm reactors (MBBR) were operated to assess pH-associated anammox activity changes during long term treatment of anaerobically digested sludge centrate pre-treated in a suspended growth partial nitrification reactor. The pH was maintained at 6.5 in reactor R1, while it was allowed to vary naturally between 7.5 and 8.1 in reactor R2. At high nitrogen loads reactor R2 had a 61% lower volumetric specific nitrogen removal rate than reactor R1. The low pH and the associated low free ammonia (FA) concentrations were found to be critical to stable anammox activity in the MBBR. Nitrite enhanced the nitrogen removal rate in the conditions of low pH, all the way up to the investigated level of 50 mg NO₂-N/L. At low FA levels nitrite concentrations up to 250 mg NO₂-N/L did not cause inactivation of anammox consortia over a 2-days exposure time.     

基于好氧反硝化及反硝化聚磷菌强化的低温低碳氮比生活污水生物处理中试研究

【背景】低碳氮比生活污水很难达标处理,多级A/O工艺、生物强化技术及生物膜技术的有机结合可有效解决这一问题。【目的】开发出一种泥膜共生多级A/O工艺并进行中试研究,驯化出高效脱氮除磷菌剂并对系统进行生物强化。【方法】通过测定中试设备出水及污水处理厂出水化学需氧量(Chemical oxygen demand,COD)、氨氮(NH_4~+-N)、硝氮(NO_3~--N)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)对比分析两种工艺的污染物去除效能,利用高通量测序技术对比生物强化技术对系统微生物群落结构的影响。【结果】中试设备对COD、NH_4~+-N、NO_3~--N、TN、TP的去除效果均优于污水处理厂的处理工艺;驯化的低温好氧反硝化菌TN去除率最大值可达84.21%,驯化的低温反硝化聚磷菌群对磷的去除率最高可达85.75%;利用驯化菌群对中试设备进行生物强化后较好地改善了系统NH_4~+-N、NO_3~--N、TN、TP的去除效果;经生物强化后,具有好氧反硝化和反硝化聚磷功能的Pseudomonas菌群明显增多。【结论】泥膜共生多级A/O工艺对于低碳氮比生活污水的处理具有很好的效果,利用生物强化技术可有效提高低温条件下系统污染物去除效能。