厌氧氨氧化生物脱氮技术的研究进展

厌氧氨氧化是指在厌氧条件下,厌氧氨氧化混合菌直接以NH4 为电子供体,以NO3-或NO2-为电子受体,将NH4^ 、NO3-或NO2-转变成N2的过程。厌氧氨氧化作为一种新型的污水处理工艺具有较高的理论意义和良好的应用前景。本文主要阐述了厌氧氨氧化生物脱氮技术原理、厌氧氨氧化的可能途径、方法及其应用现状,并且讨论了厌氧氨氧化反应的微生物学机理和厌氧氨氧化工艺的开发,提出了今后研究的主要方向。     

厌氧氨氧化体的组成、结构与功能

厌氧氨氧化(Anammox)是微生物和环境领域的研究热点之一。厌氧氨氧化菌(AnAOB)是Anammox的功能载体。不同于大部分原核微生物,AnAOB具有独特的细胞器——厌氧氨氧化体,它是进行Anammox代谢的场所。研究厌氧氨氧化体有助于探明厌氧氨氧化菌的代谢特性。本文综述了厌氧氨氧化体的组成、结构与功能,以期为从事Anammox研究的同行提供参考。     

厌氧氨氧化细菌及其群落内细菌互作关系研究进展

厌氧氨氧化细菌具有的厌氧氨氧化反应是在厌氧条件下将氨氮和亚硝氮或一氧化氮转化为硝氮、生成氮气的过程,因其能够高效低能地处理低碳氮比废水而广受关注.目前,厌氧氨氧化细菌仍未实现纯培养,借助宏组学手段研究厌氧氨氧化细菌及其群落内细菌之间的互作关系是近年来的研究趋势.本文介绍了厌氧氨氧化细菌的种类和特性,综述了厌氧氨氧化细菌...     

厌氧氨氧化颗粒污泥聚集机制研究进展

厌氧氨氧化(anaerobic ammonium oxidation,anammox)工艺被认为是当前污水生物脱氮领域最经济的处理工艺,有利于实现污水处理厂的能源自给。厌氧氨氧化菌是该工艺的核心功能微生物。以厌氧氨氧化菌为主导微生物形成的厌氧氨氧化颗粒污泥具有沉速大、污泥持留能力强及对不利环境抵抗能力强等突出优势,是实现厌氧氨氧化工艺最有前景的污泥形态。本论文围绕厌氧氨氧化颗粒,介绍了厌氧氨氧化菌的特性、种类及代谢途径,综述了厌氧氨氧化颗粒污泥的形成假说及与厌氧氨氧化颗粒污泥聚集密切相关的胞外聚合物(extracellular polymeric substance,EPS)和群体感应研究现状,并对今后厌氧氨氧化颗粒的研究进行了展望,以期为后续厌氧氨氧化颗粒的研究及厌氧氨氧化颗粒工艺的优化提供参考。     

厌氧氨氧化菌特性及其在生物脱氮中的应用

在无分子氧环境中,同时存在NH4^＋和NO2^-时,NH4^＋作为反硝化的无机电子供体,NO2^-作为电子受体,生成氮气,这一过程称为厌氧氨氧化。目前已经发现了3种厌氧氨氧化菌（Brocadia anammoxidans,Kuenenia stuttgartiensis,Scalindua sorokinii）;对厌氧氨氧化菌的细胞色素、营养物质、抑制物、结构特征和生化反应机理的研究表明,厌氧氨氧化菌具有多种代谢能力。基于部分硝化至亚硝酸盐,然后与氨一起厌氧氨氧化,以及厌氧氨氧化菌与好氧氨氧化菌或甲烷菌的协同耦合作用,提出了几种生物脱氮的新工艺（ANAMMOX、SHARON—ANAMMOX、CANON和甲烷化与厌氧氨氧化耦合工艺）。     

海洋氮循环中细菌的厌氧氨氧化

细菌厌氧氨氧化过程是在一类特殊细菌的厌氧氨氧化体内完成的以氨作为电子供体硝酸盐作为电子受体的一种新型脱氮反应.厌氧氨氧化菌的发现,改变人们对传统氮的生物地球化学循环的认识:反硝化细菌并不是大气中氮气产生的唯一生物类群.而且越来越多的证据表明,细菌厌氧氨氧化与全球的氮物质循环密切相关,估计海洋细菌的厌氧氨氧化过程占到全球海洋氮气产生的一半左右.由于氮与碳的循环密切相关,因此可以推测,细菌的厌氧氨氧化会影响大气中的二氧化碳浓度,从而对全球气候变化产生重要影响.另外,由于细菌厌氧氨氧化菌实现了氨氮的短程转化,缩短了氮素的转化过程,因此为开发更节约能源、更符合可持续发展要求的废水脱氮新技术提供了生物学基础.     

厌氧氨氧化菌的研究进展

近年来,有关厌氧氨氧化过程这一特殊的生化机制以及微生物类群的研究引起了人们的极大关注,尤其是这类微生物的生态生境可能比人们预想的范围更加广泛,因而在自然界N循环中可能具有重要意义。对这类菌结构特征、系统发育地位以及厌氧氨氧化小体和厌氧氨氧化机制的更深入认识将大大促进它们在污水处理工程中的应用。综述了近年来有关厌氧氨氧化菌的生理特性、生化机制、结构特点、生态生境以及工程应用等方面的最新进展。     

海洋厌氧氨氧化细菌分子生态学研究进展

厌氧氨氧化细菌是能在厌氧的条件下将氨氧化为氮气的一类细菌,这类细菌执行着以前未被人们所认知的一个独特的过程--氧氨氧化过程,据估计厌氧氨氧化过程对于海洋氮气的形成有30%～50%的贡献率;海洋厌氧氨氧化细菌能与氨氧化细菌及氨氧化古菌存在潜在的耦合作用,对于海洋氮循环复杂机制的阐述有着非常重要的意义;同时海洋厌氧氨氧化细菌独特的细胞和基因组结构,也成为了解海洋细菌进化重要的模式微生物之一.本文综述了近年来国内外厌氧氨氧化细菌分子生态学方面的进展,并结合作者的工作对未来的研究进行展望.     

厌氧氨氧化菌内含物——糖原颗粒

厌氧氨氧化菌(anaerobic ammonium-oxidizing bacteria, AnAOB)是分类学上新近建立的细菌类群。由于生长缓慢，培养困难，迄今没有获得纯培物。与已知细菌类群相比，AnAOB具有诸多特异性细胞结构和功能。AnAOB是化能自养型细菌，但在其细胞内经常可见贮藏性的内含物——糖原颗粒。探讨这种糖原颗粒的性状与动态，可深化人们对AnAOB的认识。本文结合文献报道及前期研究基础，对厌氧氨氧化菌糖原颗粒的结构、代谢和功能特性进行了探讨，分析认为AnAOB糖原颗粒分布于核糖细胞质内，且处于多途径合成与多位点消耗的动态平衡中；此外，糖原颗粒具有提供能量、碳架和应激保护等能力，对逆境下AnAOB的生存具有重要意义。本综述可为厌氧氨氧化菌的深入研究和工程应用提供支撑。     

厌氧氨氧化细菌的研究进展

厌氧氨氧化是指微生物在无氧条件下,以NO_2~–为电子受体,将NH_4~+氧化成N_2的过程,该过程主要由浮霉菌门下的厌氧氨氧化细菌参与。厌氧氨氧化细菌广泛存在于海洋生态系统、淡水生态系统、陆地生态系统及其他一些特殊生境中,其在废水生物脱氮和地球氮循环中扮演着重要角色。本文从厌氧氨氧化细菌的发现历程、种类、特性、代谢途径、分布、检测方法及应用上进行了较为全面的总结;最后对厌氧氨氧化细菌研究前沿问题和未来发展方向进行了探讨与展望。     

Anaerobic ammonium oxidation in constructed wetlands with bio-contact oxidation as pretreatment

Anaerobic ammonium oxidation (ANAMMOX) may provide an effective nitrogen removal pathway for constructed wetlands with low C/N influent. In a study of domestic sewage treatment, anaerobic ammonium oxidation process was identified in the pilot-scale constructed wetland of a bio-ecological process which was composed of a bio-contact oxidation reactor and a horizontal subsurface flow constructed wetland (CW). To investigate the ANAMMOX establishment in the bio-ecological process, two new CWs (planted and unplanted) were developed to be a control for the pre-existing CW. Under operational conditions of DO 2-3 mg/l, HRT 3.5 h for the bio-contact oxidation reactor, HRT 3 days for CWs, and domestic sewage as influent, the process achieved more than 90% TN removal rate after the ANAMMOX was established. The ANAMMOX bacteria on the media of the constructed wetlands were analyzed by specific polymerase chain reaction (PCR) with ANAMMOX specific primer set AMX818F-AMX1066R. The result of the genetic sequencing showed that the PCR product was related to Candidatus B. anammoxidans (AF375994.1) with 98% sequence similarity. Copy numbers of 16S rRNA gene of ANAMMOX bacteria in the pre-existing CW, the new planted CW and new unplanted CW were 3.47 × 10⁵, 3.02 × 10⁵ and 1.30 × 10⁵, respectively. These results demonstrated that the ANAMMOX process was successfully established and operated consistently in the constructed wetlands with a bio-contact oxidation reactor as a pretreatment, and that vegetation positively affected the growth and enrichment of ANAMMOX bacteria.     

厌氧氨氧化污泥中效应菌的分子生物学研究

对具有厌氧氨氧化作用的细菌进行更深入的分析和了解有助于该新型生物脱氮过程在实践中的应用,采用分子生物学方法从已培养的具有厌氧氨氧化活性的污泥中提取细菌总DNA,经纯化、特异引物PCR扩增、克隆、测序等过程,得到厌氧氨氧化菌部分16S rDNA序列(长度为836bp),少部分克隆具有1～2个碱基的突变。此外,进化分析结果显示培养获得的细菌与已发现的Candidatus Brocadia anammoxidans、Anaerobic ammoniumoxidizing Planctomycete、Uncultured anoxic sludge bacterium KU1细菌在进化上关系较近,但比对分析结果表明所研究的细菌与上述细菌的DNA序列相似度不高,这表明自然环境中还存在一种以前未被发现的可进行厌氧氨氧化的细菌。     

厌氧氨氧化工艺的应用现状和问题

厌氧氨氧化(Anaerobic ammonium oxidation,ANAMMOX)工艺因其高效低耗的优势,在废水生物脱氮领域具有广阔的应用前景。在过去的20年中,许多基于ANAMMOX反应的工艺得以不断研究和应用。预计到2014年末,全球范围内的ANAMMOX工程将会超过100座。综述了各种形式的ANAMMOX工艺,包括短程硝化-厌氧氨氧化、全程自养脱氮、限氧自养硝化反硝化、反硝化氨氧化、好氧反氨化、同步短程硝化-厌氧氨氧化-反硝化耦合、单级厌氧氨氧化短程硝化脱氮工艺。对一体式和分体式工艺运行条件进行了比较,结合ANAMMOX工艺工程(主要包括移动床生物膜,颗粒污泥和序批式反应器系统)应用现状,总结了工程化应用过程中遇到的问题及其解决对策,在此基础上对今后的研究和应用方向进行了展望。今后的研究重点应集中于运行条件的优化和水质障碍因子的解决,尤其是工艺自动化控制系统的开发和特殊废水对工艺性能影响的研究。     

Inactivation of ANAMMOX communities under concurrent operation of anaerobic ammonium oxidation (ANAMMOX) and denitrification

A concurrent operation of anaerobic ammonium oxidation (ANAMMOX) and denitrification was investigated in a well known UASB reactor seeding with both ANAMMOX and anaerobic granular sludges. ANAMMOX activity was confirmed by hydroxylamine test and the hybridization of biomass using the gene probes of Amx 820 and EUB 338 mixed. Denitrification was observed through the reductions of both COD and nitrate-nitrite concentrations under anaerobic/anoxic conditions. By providing a stoichiometric ratio of nitrite to ammonium nitrogen with addition nitrate nitrogen, a gradual reduction of ANAMMOX activity was found with an increase of COD concentration in a range of 100-400 mg l(-1). This is equivalent to the COD to N ratio of 0.9-2.0. The COD concentration was found to be a control variable for process selection between ANAMMOX reaction and denitrification. A reduction of COD and nitrite-nitrate concentrations in all reactors confirmed the undergone concurrent denitrification which thrives when sufficient organic matter is available. COD concentration over 300 mg l(-1) was found to inactivate or eradicate ANAMMOX communities.     

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.     

Batch culture enrichment of ANAMMOX populations from anaerobic and aerobic seed cultures

Discharge of nitrate and ammonia rich wastewaters into the natural waters encourage eutrophication, and contribute to aquatic toxicity. Anaerobic ammonium oxidation process (ANAMMOX) is a novel biological nitrogen removal alternative to nitrification-denitrification, that removes ammonia using nitrite as the electron acceptor. The feasibility of enriching the ANAMMOX bacteria from the anaerobic digester sludge of a biomethanation plant treating vegetable waste and aerobic sludge from an activated sludge process treating domestic sewage is reported in this paper. ANAMMOX bacterial activity was monitored and established in terms of nitrogen transformations to ammonia, nitrite and nitrate along with formation of hydrazine and hydroxylamine.     

Characteristics of aerobic and anaerobic ammonium-oxidizing bacteria in the hyporheic zone of a contaminated river

Both β-proteobacterial aerobic ammonium-oxidizing bacteria (AOB) and anaerobic ammonium-oxidizing (ANAMMOX) bacteria were investigated in the hyporheic zone of a contaminated river in China containing high ammonium levels and low chemical oxygen demand. Fluorescence in-situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE) and cloning-sequencing were employed in this study. FISH analysis illustrated that AOB (average population of 3.5?%) coexisted with ANAMMOX bacteria (0.7?%). The DGGE profile revealed a high abundance and diversity of bacteria at the water-air-soil interface rather than at the water-soil interface. The redundancy analysis correlated analysis showed that the diversity of ANAMMOX bacteria was positively related to the redox potential. The newly detected sequences of ANAMMOX organisms principally belonged to the genus Candidatus "Brocadia", while most ammonia monooxygenase subunit-A gene amoA sequences were affiliated with Nitrosospira and Nitrosomonas. These results suggest that the water-air-soil interface performs an important function in the nitrogen removal process and that the bioresources of AOB and ANAMMOX bacteria can potentially be utilized for the eutrophication of rivers.     

ANAMMOX process start up and stabilization with an anaerobic seed in Anaerobic Membrane Bioreactor (AnMBR)

ANaerobic AMMonium OXidation (ANAMMOX) process, an advanced biological nitrogen removal alternative to traditional nitrification--denitrification removes ammonia using nitrite as the electron acceptor without oxygen. The feasibility of enriching anammox bacteria from anaerobic seed culture to start up an Anaerobic Membrane Bioreactor (AnMBR) for N-removal is reported in this paper. The Anammox activity was established in the AnMBR with anaerobic digester seed culture from a Sewage Treatment Plant in batch mode with recirculation followed by semi continuous process and continuous modes of operation. The AnMBR performance under varying Nitrogen Loading Rates (NLR) and HRTs is reported for a year, in terms of nitrogen transformations to ammoniacal nitrogen, nitrite and nitrate along with hydrazine and hydroxylamine. Interestingly ANAMMOX process was evident from simultaneous Amm-N and nitrite reduction, consistent nitrate production, hydrazine and hydroxylamine presence, notable organic load reduction and bicarbonate consumption.     

Quantitative comparison of stability of ANAMMOX process in different reactor configurations

A new stability evaluating system for ANAMMOX comprising three instability indices i.e. coefficient of variation ratio, coefficient of range ratio and coefficient of regression function derivative was established. Three lab-scale ANAMMOX reactors viz upflow anaerobic sludge blanket (UASB) reactor, upflow stationary fixed film (USFF) reactor and anaerobic sequencing batch reactor (ASBR) were compared for their stability based on the established criterion against the hydraulic and substrate concentration shocks. The results showed that all ANAMMOX reactors under investigation were more tolerant to the hydraulic shock than substrate concentration shock. The UASB reactor was the most stable reactor configuration towards substrate concentration shock, followed by the USFF reactor and ASBR. However, the ASBR proved the most tolerant to hydraulic shock, followed by the UASB reactor and USFF reactor. In terms of stability, UASB reactor was more suitable configuration compared with USFF reactor. The instability indices proved to be effective and explicit for the evaluation of ANAMMOX systems.