Combined anaerobic ammonium and methane oxidation for nitrogen and methane removal

Anammox (anaerobic ammonium oxidation) is an environment-friendly and cost-efficient nitrogen-removal process currently applied to high-ammonium-loaded wastewaters such as anaerobic digester effluents. In these wastewaters, dissolved methane is also present and should be removed to prevent greenhouse gas emissions into the environment. Potentially, another recently discovered microbial pathway, n-damo (nitrite-dependent anaerobic methane oxidation) could be used for this purpose. In the present paper, we explore the feasibility of simultaneously removing methane and ammonium anaerobically, starting with granules from a full-scale anammox bioreactor. We describe the development of a co-culture of anammox and n-damo bacteria using a medium containing methane, ammonium and nitrite. The results are discussed in the context of other recent studies on the application of anaerobic methane- and ammonia-oxidizing bacteria for wastewater treatment.     

Spatial separation of anaerobic ammonium oxidation and nitrite-dependent anaerobic methane oxidation in permeable riverbeds

Anaerobic ammonium oxidation (anammox) and nitrite-dependent anaerobic methane oxidation (n-damo) play important roles in nitrogen and carbon cycling in fresh waters but we do not know how these two processes compete for their common electron acceptor, nitrite. Here, we investigated the spatial distribution of anammox and n-damo across a range of permeable riverbed sediments. Anammox activity and gene abundance were detected in both gravel and sandy riverbeds and showed a simple, common vertical distribution pattern, while the patterns in n-damo were more complex and n-damo activity was confined to the more reduced, sandy riverbeds. Anammox was most active in surficial sediment (0–2 cm), coincident with a peak in hzsA gene abundance and nitrite. In contrast, n-damo activity peaked deeper down (4–8 cm) in the sandy riverbeds, coincident with a peak in n-damo 16S rRNA gene abundance and higher methane concentration. Pore water nitrite, methane and oxygen were key factors influencing the distribution of these two processes in permeable riverbeds. Furthermore, both anammox- and n-damo- activity were positively correlated with denitrification activity, suggesting a role for denitrification in supplying both processes with nitrite. Our data reveal spatial separation between anammox and n-damo in permeable riverbed sediments that potentially avoids them competing for nitrite.     

Simultaneous nitrite-dependent anaerobic methane and ammonium oxidation processes

Nitrite-dependent anaerobic oxidation of methane (n-damo) and ammonium (anammox) are two recently discovered processes in the nitrogen cycle that are catalyzed by n-damo bacteria, including "Candidatus Methylomirabilis oxyfera," and anammox bacteria, respectively. The feasibility of coculturing anammox and n-damo bacteria is important for implementation in wastewater treatment systems that contain substantial amounts of both methane and ammonium. Here we tested this possible coexistence experimentally. To obtain such a coculture, ammonium was fed to a stable enrichment culture of n-damo bacteria that still contained some residual anammox bacteria. The ammonium supplied to the reactor was consumed rapidly and could be gradually increased from 1 to 20 mM/day. The enriched coculture was monitored by fluorescence in situ hybridization and 16S rRNA and pmoA gene clone libraries and activity measurements. After 161 days, a coculture with about equal amounts of n-damo and anammox bacteria was established that converted nitrite at a rate of 0.1 kg-N/m(3)/day (17.2 mmol day(-1)). This indicated that the application of such a coculture for nitrogen removal may be feasible in the near future.     

水生生态系统中金属依赖型甲烷厌氧氧化过程的研究进展

甲烷是一种比CO_2更活跃的温室气体,微生物驱动的甲烷厌氧氧化(anaerobicoxidationof methane,AOM)过程对于降低全球甲烷的排放有着重要意义。参与AOM反应的最终电子受体主要分为三类,即硫酸盐、亚硝酸盐/硝酸盐以及以Fe(III)、Mn(IV)等为代表的金属离子。可溶性金属物质和不溶性金属矿物都可以被用作AOM的电子受体,这大大提高了参与金属依赖型甲烷厌氧氧化(metal-dependent anaerobic oxidation of methane,Metal-AOM)微生物的生态价值。目前研究聚焦在功能菌群、生态分布等方面。部分甲烷厌氧氧化古菌(anaerobic methanotrophic archaea,ANME)具有直接或间接参与Metal-AOM过程的能力。但由于功能菌群纯化富集和分离具有一定难度,有关其生理生化和生态学等特征的研究受到限制。同时,随着Metal-AOM被发现存在于不同水生生境中,其在污染治理领域的应用也被广泛讨论,但是河口生境尚缺乏深入研究。本文从Metal-AOM的发现入手,阐述了参与该过程的主要微生物及其在水域环境下的生态分布,并介绍了Metal-AOM的反应机制和在实际应用中的机遇与挑战。最后,根据现有研究结果,提出对功能菌群、机制及环保应用的研究展望,包括微生物分离纯化和影响因素、菌群代谢活性和作用机制的解析以及新型生产工艺的设计和发展应用,以期为今后的环境污染治理和工业应用提供借鉴意义。     

Nitrogen source effects on the denitrifying anaerobic methane oxidation culture and anaerobic ammonium oxidation bacteria enrichment process

The co-culture system of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) has a potential application in wastewater treatment plant. This study explored the effects of permutation and combination of nitrate, nitrite, and ammonium on the culture enrichment from freshwater sediments. The co-existence of NO₃ ⁻, NO₂ ⁻, and NH₄ ⁺ shortened the enrichment time from 75 to 30 days and achieved a total nitrogen removal rate of 106.5 mg/L/day on day 132. Even though ammonium addition led to Anammox bacteria increase and a higher nitrogen removal rate, DAMO bacteria still dominated in different reactors with the highest proportion of 64.7% and the maximum abundance was 3.07 ± 0.25 × 10⁸ copies/L (increased by five orders of magnitude) in the nitrite reactor. DAMO bacteria showed greater diversity in the nitrate reactor, and one was similar to M. oxyfera; DAMO bacteria in the nitrite reactor were relatively unified and similar to M. sinica. Interestingly, no DAMO archaea were found in the nitrate reactor. This study will improve the understanding of the impact of nitrogen source on DAMO and Anammox co-culture enrichment.

     

白洋淀富营养化湖泊湿地厌氧氨氧化菌的分布及对氮循环的影响

随着生物反应器和海洋生态系统中厌氧氨氧化反应的发现,自然生态系统的氮循环过程被重新认识,但是目前厌氧氨氧化过程是否也存在于富营养化湖泊湿地并发挥着重要作用,还未见报道。结合15N同位素示踪与分子生物学技术研究了白洋淀湖泊湿地沉积物中厌氧氨氧化菌的分布、菌群结构特性、生物多样性及其活性。结果表明,在藻类影响导致的高氨氮沉积物中,厌氧氨氧化菌具有广泛存在性。通过构建16S rRNA克隆文库发现,沉积物中厌氧氨氧化菌的生物多样性相对较低,在2%差异度的条件下,30个克隆序列只分为5个操作分类单元(OTUs),代表序列与Genebank数据库中已探明的厌氧氨氧化菌Candidatus Brocadia fulgida和Candidatus Brocadia anammoxidans同源性最高,分别可达97%和96%。同位素示踪结果表明,白洋淀湖泊湿地沉积物中厌氧氨氧化活性为0.19—7.78 nmol N g-1h-1,空间差异较大,产生的氮气占此沉积物总氮气生成量的0.64%—20.65%,体现了湿地的异质性。通过得出的厌氧氨氧化反应速率推算每年由厌氧氨氧化反应损失的氮量为1.8—78gN m-2a-1,对白洋淀氮循环起到非常重要的作用。富营养化湖泊湿地沉积物中厌氧氨氧化反应的发现为研究厌氧氨氧化对氮循环的重要影响提供了新证据。     

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。     

自然生态系统中的厌氧氨氧化

厌氧氨氧化（anaerobic ammonium oxidation,anammox）是由anammox菌在缺氧条件下以氨为电子供体、以亚硝酸为电子受体的生物反应,反应产物为氮气,该反应的发现为全球氮素循环增添了新的内容。参与anammox反应的微生物是anammox菌,anammox菌是一群分支很深的浮霉状菌,目前已发现的anammox菌有5个属8个种。催化anammox反应的是一特殊的细胞结构-厌氧氨氧化体,每种已发现的anammox菌中都存在该特殊结构。有关anammox反应的生化机理目前普遍认为,NO和联氨（N2H4）是anammox反应的重要中间体,NO可将NH4 直接氧化,形成N2H4,N2H4在联氨氧化酶的作用下最终转化为氮气。Anammox最初发现于人工脱氮系统,已发现的8种anammox菌中7种来自于人工系统。但越来越多的证据表明,anammox菌广泛分布于自然界的海洋、淡水和陆地生态系统中,在区域氮素循环中起着不同程度的作用。影响自然生态系统中anammox反应的主要环境因子包括有机质含量、NO3-浓度和盐度等,但在不同的生态系统,anammox反应的主导影响因子存在较明显差异。本文综述了anammox菌的类群和生化反应机理,总结了anammox菌在各种自然生态系统中的分布与生态多样性,并论述了anammox反应在全球氮素循环中的重要性以及影响此过程发挥的主要环境因子。     

湿地微生物介导的甲烷排放机制

湿地生态系统是陆地上巨大的有机碳库,同时也是大气中甲烷(CH_4)的主要排放源。由于CH_4对全球的增温潜能是CO2的34倍,因此关于湿地CH_4排放在全球气候变化中有关碳汇、碳源的研究具有极其重要的意义。全球80%–90%的CH_4排放离不开微生物活动,湿地生态系统中产CH_4菌和CH_4氧化菌的种类组成、数量及功能与CH_4通量密切相关,但基于湿地生态系统中介导CH_4循环的功能微生物对甲烷排放通量的影响及作用机制研究相对比较分散。为更好地认识微生物介导的CH_4排放过程的微生物调控机制,本文综述了湿地生态系统中参与CH_4循环的功能微生物,对介导CH_4循环相关微生物活性的影响因子进行了回顾,重点总结了湿地生态系统微生物介导的CH_4排放机制,并对未来的相关研究方向进行了展望。由于湿地微生物介导的碳循环过程也可能决定了湿地生态系统对全球气候变暖的反馈,因此本文也能为全球气候变化研究提供微生物方面的参考。     

Phylogenetic analysis of a microbial community involved in anaerobic oxidation of ammonium nitrogen

The phylogenetic diversity of a microbial community involved in anaerobic oxidation of ammonium nitrogen in the DEAMOX process was studied. Analysis of clone libraries containing 16S rRNA gene inserts of Bacteria, (including Planctomycetes) and Archaea revealed the presence of nucleotide sequences of the microorganisms involved in the main reactions of the carbon, nitrogen, and sulfur cycles, including nitrifying, denitrifying, and ANAMMOX bacteria. In the bacterial clone library, 16S rRNA gene sequences of representatives of the phyla Proteobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Verrucomicrobia, Lentisphaerae, Spirochaetales, and Planctomycetes, as well as of some new groups, were detected. In the archaeal clone library, nucleotide sequences of methanogens belonging to the orders Methanomicrobiales, Methanobacteriales, and Methanosarcinales were found. It is possible that both ANAMMOX bacteria and bacteria of the genus Nitrosomonas are involved in anaerobic ammonium oxidation in the DEAMOX reactor. Many sequences were similar to those from the clone libraries obtained previously from the ANAMMOX community of marine sediments. It is also probable that the DEAMOX reactions occur in natural ecosystems (in marine and freshwater sediments and the oceanic water column), thereby providing for the coupling of the nitrogen and sulfur cycles.     

红树林生态系统微生物驱动的氮素循环过程研究进展

微生物驱动的氮循环过程在红树林生态系统物质循环、净化外来污染物、维持生态系统平衡等方面起重要作用。相较于其他自然生态系统,因红树林处于沿海陆地交界地带,其氮循环过程及其相关微生物的种类丰富,受交错复杂的环境因素影响与调控。本文梳理了红树林土壤性质及特性,综述了红树林生态系统中由微生物驱动的固氮、氮素矿化、硝化、厌氧氨氧化、反硝化、异化硝酸盐还原为铵等主要的氮循环过程,并讨论了氮循环与其他循环的耦合过程。最后讨论pH、盐度、季节、螃蟹活动、红树林树种等环境因素对氮循环过程及其相关微生物丰度、多样性的影响。本综述以期为红树林湿地生态系统的保护和修复提供理论参考。     

厌氧氨氧化菌群体感应系统研究

厌氧氨氧化(Anammox)是以铵为电子供体将亚硝酸盐转化为氮气的生物过程。厌氧氨氧化菌(AAOB)生理代谢和细胞结构均十分特殊,且在氮素循环中起着十分重要的作用。厌氧氨氧化已成为环境学、微生物学、海洋学等领域的研究热点。但是,至今人们未能对厌氧氨氧化菌进行纯培养,这严重限制了对厌氧氨氧化菌的深入研究。群体感应是一种普遍存在于微生物细胞之间的通讯机制,它具有根据菌群密度和周围环境变化调节基因表达,以控制细菌群体行为的功能。厌氧氨氧化菌活性的细胞密度效应和生物团聚行为与细菌中普遍存在的群体感应现象相符。探讨了厌氧氨氧化菌群体感应系统存在的可能性、工作机制及其生态学意义,以期为厌氧氨氧化菌的分离培养、团聚体培育等提供理论指导。     

Enhancing mainstream nitrogen removal by employing nitrate/nitrite-dependent anaerobic methane oxidation processes

Due to serious eutrophication in water bodies, nitrogen removal has become a critical stage for wastewater treatment plants (WWTPs) over past decades. Conventional biological nitrogen removal processes are based on nitrification and denitrification (N/DN), and are suffering from several major drawbacks, including substantial aeration consumption, high fugitive greenhouse gas emissions, a requirement for external carbon sources, excessive sludge production and low energy recovery efficiency, and thus unable to satisfy the escalating public needs. Recently, the discovery of anaerobic ammonium oxidation (anammox) bacteria has promoted an update of conventional N/DN-based processes to autotrophic nitrogen removal. However, the application of anammox to treat domestic wastewater has been hindered mainly by unsatisfactory effluent quality with nitrogen removal efficiency below 80%. The discovery of nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) during the last decade has provided new opportunities to remove this barrier and to achieve a robust system with high-level nitrogen removal from municipal wastewater, by utilizing methane as an alternative carbon source. In the present review, opportunities and challenges for nitrate/nitrite-dependent anaerobic methane oxidation are discussed. Particularly, the prospective technologies driven by the cooperation of anammox and n-DAMO microorganisms are put forward based on previous experimental and modeling studies. Finally, a novel WWTP system acting as an energy exporter is delineated.     

厌氧氨氧化组合脱氮工艺研究进展

厌氧氨氧化工艺是一项高效、低耗的生物脱氮工艺,但受限于底物类型、硝氮积累等问题,其在主流应用中仍然面临一些挑战。近些年来,针对上述问题,厌氧氨氧化组合工艺得到了广泛关注。通过对近年来所开发的厌氧氨氧化组合工艺,从工艺原理、优缺点、影响因素、工艺拓展性及其在推广应用中存在的关键瓶颈等角度进行探讨,并结合课题组相关工作,展望了厌氧氨氧化组合工艺在城市生活污水处理中的发展前景。     

Key physiology of anaerobic ammonium oxidation.

The physiology of anaerobic ammonium oxidizing (anammox) aggregates grown in a sequencing batch reactor was investigated quantitatively. The physiological pH and temperature ranges were 6.7 to 8.3 and 20 to 43 degrees C, respectively. The affinity constants for the substrates ammonium and nitrite were each less than 0.1 mg of nitrogen per liter. The anammox process was completely inhibited by nitrite concentrations higher than 0.1 g of nitrogen per liter. Addition of trace amounts of either of the anammox intermediates (1. 4 mg of nitrogen per liter of hydrazine or 0.7 mg of nitrogen per liter of hydroxylamine) restored activity completely.     

Nitrogen removal with the anaerobic ammonium oxidation process

Anaerobic ammonium-oxidizing (anammox) bacteria convert ammonium to N₂ with nitrite as the terminal electron acceptor in the absence of O₂. Nitritation–anammox bioreactors provide a cost-effective and environment-friendly alternative to conventional nitrification/denitrification nitrogen removal systems. Currently, this process is only applied for ammonium removal from wastewater with high ammonium load and temperature. Nevertheless, recent results obtained with laboratory-scale bioreactors suggest new possible routes of application of the Nitritation–anammox technology including (1) municipal wastewater treatment, removal of (2) methane in combination with nitrite-reducing methane-oxidizing bacteria, (3) nitrate coupled to organic acid oxidation and (4) nitrogen oxides. The current review summarizes the state-of-the-art of the application of Nitritation–anammox systems and discusses the possibilities of utilizing these recent results for wastewater treatment.     

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NO2-为电子受体,将NH4+氧化成N2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、...