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。     

中试厌氧氨氧化反应器的启动与调控

研究了中试厌氧氨氧化(Anaerobic ammonium oxidation,Anammox)反应器的启动性能。结果表明,以硝化反硝化污泥、短程硝化污泥、厌氧絮体污泥和厌氧颗粒污泥混合接种,经过255d的运行,可在常温下(5oC～27oC)成功启动中试Anammox反应器,反应器的基质氮去除速率可达1.30kg/(m3·d)。厌氧氨氧化是致碱反应,厌氧氨氧化成为反应器内的主导反应后,进水pH宜控制在厌氧氨氧化适宜范围的偏低水平(6.8左右)。亚硝酸盐既是Anammox菌的基质,也是抑制剂,控制进水亚硝酸盐浓度(13～36mg/L)有助于厌氧氨氧化反应。菌种是生物反应器的功能之源,向中试装置投加少量厌氧氨氧化污泥(投加比2%),可大大加速中试Anammox反应器的启动进程。     

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。     

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。     

污水处理系统中厌氧氨氧化菌分布及影响因素

基于厌氧氨氧化的污水生物脱氮工艺近年来发展迅速,污水处理系统中厌氧氨氧化菌的分布和多样性成为了重要的研究方向。目前,在污水处理系统中曾检测出多种厌氧氨氧化(Anaerobic ammonium oxidation,Anammox)菌,最常被检测出的是待定布罗卡地菌Candidate Brocadia和待定斯图加特库氏菌Candidate Kuenenia的Anammox菌,并且研究发现单一生境下往往只存在一种类型的Anammox菌,但是影响Anammox菌分布和多样性的因素与机制目前仍不明确。系统总结了污水处理系统中,不同工艺形式和运行条件下的Anammox菌分布情况,归纳分析了关键因素对Anammox菌分布的影响,包括底物浓度和微生物比生长速率、污泥性质与微生物生境、多重因素的联合作用和影响等。在此基础上,阐述了Anammox菌分布机制研究的工程意义,并对该领域的研究方向和思路进行了展望。     

三种接种物启动Anammox-EGSB反应器的性能

为了优选接种物和加速Anammox反应器启动,分别以厌氧产甲烷污泥 (Anaerobic methanogenic sludge,AMS)、新鲜厌氧氨氧化污泥 (Fresh Anammox sludge,FAS) 和储藏厌氧氨氧化污泥 (Stored Anammox sludge,SAS) 作为接种物,研究了厌氧氨氧化膨胀颗粒污泥床 (Anammox-EGSB) 反应器 (R1、R2和R3) 的启动性能。结果表明：3种接种物均能成功启动Anammox-EGSB反应器,启动性能的优劣次序为：R2 (接种物为     

厌氧氨氧化颗粒污泥研究进展

厌氧氨氧化(Anaerobic ammonium oxidation,Anammox)工艺是一种新的生物脱氮技术。一经问世即得到人们青睐,现已成为废水脱氮的升级技术。厌氧氨氧化菌(Anaerobicammoniumoxidation bacteria,AnAOB)是Anammox工艺的功能之源。以颗粒污泥形态存在的AnAOB是Anammox颗粒污泥床脱氮系统的重要支柱。由于AnAOB生长缓慢且对环境条件变化敏感,Anammox脱氮系统不仅启动缓慢,而且运行极易失稳甚至崩溃。值得庆幸的是,AnAOB可自主选择、组合和固定功能菌群落而形成Anammox颗粒污泥,并通过其优良的重力沉降性能和高效的基质转化性能保障Anammox脱氮系统的持续工作。本文综述了AnAOB的种类和特性及Anammox颗粒污泥的组成、结构和功能,以期为Anammox工艺的优化和拓展提供参考。     

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.     

厌氧氨氧化反应器启动过程的影响因素及微生物群落变化研究进展

自厌氧氨氧化反应发现以来,由于其具有低能耗、无需外加碳源等优点,已成为人们在污水生物脱氮研究与应用中的最新关注点。然而,由于极低的生长速率、极长的倍增时间以及严格的代谢条件等特点,限制了厌氧氨氧化菌的应用。综述了厌氧氨氧化菌富集培养过程中的影响因素,介绍了不同污泥来源的厌氧氨氧化优势菌属、分子鉴定方法,提供了部分用于厌氧氨氧化菌鉴定使用的引物序列和厌氧氨氧化菌最新发现的属与种。最后,对未来的研究方向提出一些建议思考,以期为厌氧氨氧化工艺在污水处理中的应用提供参考。     

Inhibitory effects of free ammonia on Anammox bacteria

Anammox bacteria can effectively treat high ammonia and nitrite concentrations under anoxic environments. However, the presence of high ammonia and nitrite concentrations may cause free ammonia and nitrous acid inhibition at high pH and temperature environments. In this study, the inhibitory effect of free ammonia on Anammox bacteria was investigated in a lab-scale upflow fixed-bed reactor with Kaldnes biofilm carriers. Results of continuous operation showed that inhibition was not observed in the Anammox reactor when the free ammonia concentration gradually increased up to 150?mg/L. However, Anammox activity suddenly dropped to 10 % when the free ammonia concentration reached to 190?mg/L. Nevertheless, high influent ammonia and nitrite concentrations up to 1,500 and 500?mg/L, respectively, did not noticeably inhibit the Anammox activity. Gradually decreasing Anammox activity was also supported by fluorescent in situ hybridization (FISH) analysis. FISH and 16S rRNA gene analysis results revealed that main Anammox organisms were phylogenetically related to Candidatus Kuenenia stuttgartiensis, Candidatus Jettenia asiatica and Candidatus Brocadia anammoxidans.     

厌氧铵氧化过程中关键酶及相关分子标记在生态学研究中的应用进展

厌氧铵氧化是由微生物介导的氮素循环过程中的重要途径之一。近20年来,通过对厌氧铵氧化细菌生态学、基因组学和生理代谢特性的探索,人们对其微生物学机制已经有了较多的认识:厌氧铵氧化细菌通过亚硝酸盐还原酶将亚硝酸根离子还原为一氧化氮,进而与铵离子结合在联氨合成酶的作用下生成联氨,最后通过联氨氧化酶的催化产生终产物氮气。同时,对参与这些过程的关键酶及其功能基因的认识有助于选择新的分子标记,从而为研究厌氧铵氧化细菌的多样性和分子生态学特征提供新的工具,以弥补16S rRNA基因特异性相对较低且难以与生态功能关联等方面的不足。对目前已知的参与厌氧铵氧化过程的3种关键酶的研究历程和现状进行了评述,并总结了利用3种功能基因进行厌氧铵氧化细菌生态学研究的最新进展。     

16S rRNA gene and lipid biomarker evidence for anaerobic ammonium-oxidizing bacteria (anammox) in California and Nevada hot springs

Anammox, the oxidation of ammonium with nitrite to dinitrogen gas under anoxic conditions, is an important process in mesophilic environments such as wastewaters, oceans and freshwater systems, but little is known of this process at elevated temperatures. In this study, we investigated anammox in microbial mats and sediments obtained from several hot springs in California and Nevada, using geochemical and molecular microbiological methods. Anammox bacteria-specific ladderane core lipids with concentrations ranging between 0.3 and 52 ng g⁻¹ sediment were detected in five hot springs analyzed with temperatures up to 65 °C. In addition, 16S rRNA gene analysis showed the presence of genes phylogenetically related to the known anammox bacteria Candidatus Brocadia fulgida, Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis (96.5–99.8% sequence identity) in three hot springs with temperatures up to 52 °C. Our data indicate that anammox bacteria may be able to thrive at thermophilic temperatures and thus may play a significant role in the nitrogen cycle of hot spring environments.     

Anammox反应器启动前后微生物群落结构差异性

接种A^2/O回流污泥启动Anammox-UASB反应器,研究了上升流速对系统脱氮性能影响,利用高通量测序对反应器中微生物群落结构变化进行了研究。结果表明,历时35 d成功启动Anammox反应器。上升流速升高可以明显促进脱氮效果,在最佳上升流速为1.14 m/h时TN去除率达84.74%,去除速率高达0.766 gTN/(L·d)。高通量分析表明,Anammox污泥群落Alpha多样性较接种污泥明显减少,Anammox污泥中的Anammox菌主要为Candidatus Jettenia和Candidatus Brocadia两个属,同时检测到大量的其他脱氮微生物菌属,系统中这些脱氮微生物的大量增值使系统脱氮能力逐步提高。     

高通量测序分析多种典型生境中厌氧氨氧化细菌的多样性分布特征

【背景】厌氧氨氧化过程是氮素循环过程的重要途径之一,在氮素循环中发挥重要作用。先前的研究已经证实了厌氧氨氧化细菌存在于多种生境中,但对其多样性分布还没有系统的研究。【目的】对厌氧氨氧化细菌在不同类型生境中的多样性分布规律进行深入分析,充分展示其在不同生境中的群落结构特点,并揭示多样性分布与环境因素之间的关系。【方法】在建立厌氧氨氧化细菌16S rRNA基因序列数据库的基础上,运用高通量测序技术分析其在不同生境中的多样性分布特征。【结果】厌氧氨氧化细菌在红树林、海湾和河口生境中的多样性水平较高,而污泥和红壤的多样性水平明显较低。系统发育分析表明,这些生境中的厌氧氨氧化细菌主要由Candidatus Brocadia、Ca.Scalindua和未明确分类地位的菌属组成;从河流到红树林生态系统,随着盐度的增加,厌氧氨氧化细菌的优势种属由Ca. Brocadia转变到Ca. Scalindua,相关性分析也表明了盐度是导致不同生境中厌氧氨氧化细菌群落结构差异的主要因素。【结论】不同生境中存在不同的厌氧氨氧化细菌种群结构,环境条件的差异影响了厌氧氨氧化细菌的种群分布和系统演化。     

Effects of Environmental Factors on Anammox Bacterial Community Structure in Sediments of a Freshwater Aquaculture Farm,Yangcheng Lake

Previous studies have reported wide distribution of anaerobic ammonia oxidation (anammox) bacteria in various ecosystems. However, little is known about the distribution of anammox bacteria under varying environmental conditions in intensive aquaculture systems. In Yangcheng Lake, a famous crab farm situated in the Yangtze River Delta, sediment samples were collected in October (feeding period) and January (nonfeeding period) to analyze the distribution and diversity of anammox bacteria and their relationships with environmental factors. Based on the functional biomarker of Anammox bacteria, hzo gene, anammox bacterial clone libraries were constructed and their abundances were determined by quantitative PCR (qPCR). The Anammox bacteria were detected in the lake with the abundances ranging from 0.70 × 10⁵ to 6.05 × 10⁵ copies per gram of sediment. Sequences from eight clone libraries yielded seven unique operational taxonomic units (OTUs), distantly related to the Candidatus Jettenia genera with a similarity of about 91%. The Anammox bacterial community structures, diversities and abundances varied spatiotemporally with environmental conditions. In October, the level of the nitrogen compounds, the diversity, evenness and abundance of Anammox bacteria were higher than in January. The predominant OTU of samples changed from HZO-OTU-1 (34.25%) in January to HZO-OTU-2 (28.90%) in October. Moreover, the site (SW) nearing to sewage inlet was lack of HZO-OTU-7 in January. Canonical correspondence analysis (CCA) showed that the pore water NO₂^? concentration, ammonium to nitrogen oxides ratio (NH₄⁺/NO_x^?) and total organic carbon to total nitrogen ratio (TOC/TN) contributed most to Anammox bacterial community structures variances. Pearson correlations analysis revealed that the Anammox bacteria abundance had positive co-relationships with TN, NH₄⁺, NO₃^? concentrations, and negative correlation with TOC/TN in porewater.     

Characterization of functional microbial community in a membrane-aerated biofilm reactor operated for completely autotrophic nitrogen removal

The 16S rDNA-based molecular technique was applied to investigate the functional microbial community of a membrane-aerated biofilm (MAB) that was used for completely autotrophic nitrogen removal over nitrite (CANON). The relationships among two kinds of key bacteria responsible for CANON: aerobic ammonia-oxidizing bacteria (AOB) and Anammox bacteria, and their possible distributions in the MAB were discussed based on the microbial community analysis. FISH analysis showed the existence of two visible active layers in experimental MAB. One is the partial nitrifying layer located in the region of oxygen-rich membrane-biofilm interface, dominated by NSO190-positive AOB. The other is the Anammox active layer located in the region of anoxic liquid-biofilm interface, dominated by PLA46 and AMX820-positive Anammox microorganisms. As a result of this study, the AOB as well as Anammox bacteria were present and active in experimental MABR, and the cooperation between AOB and Anammox bacteria was considered to be responsible for CANON.     

Candidatus "Scalindua brodae", sp. nov., Candidatus "Scalindua wagneri", sp. nov., two new species of anaerobic ammonium oxidizing bacteria

Anaerobic ammonium oxidation (anammox) is both a promising process in wastewater treatment and a long overlooked microbial physiology that can contribute significantly to biological nitrogen cycling in the world's oceans. Anammox is mediated by a monophyletic group of bacteria that branches deeply in the Planctomycetales. Here we describe a new genus and species of anaerobic ammonium oxidizing planctomycetes, discovered in a wastewater treatment plant (wwtp) treating landfill leachate in Pitsea, UK. The biomass from this wwtp showed high anammox activity (5.0 +/- 0.5 nmol/mg protein/min) and produced hydrazine from hydroxylamine, one of the unique features of anammox bacteria. Eight new planctomycete 16S rRNA gene sequences were present in the 16S rRNA gene clone library generated from the biomass. Four of these were affiliated to known anammox 16S rRNA gene sequences, but branched much closer to the root of the planctomycete line of descent. Fluorescence in situ hybridization (FISH) with oligonucleotide probes specific for these new sequences showed that two species (belonging to the same genus) together made up > 99% of the planctomycete population which constituted 20% of the total microbial community. The identification of these organisms as typical anammox bacteria was confirmed with electron microscopy and lipid analysis. The new species, provisionally named Candidatus "Scalindua brodae" and "Scalindua wagneri" considerably extend the biodiversity of the anammox lineage on the 16S rRNA gene level, but otherwise resemble known anammox bacteria. Simultaneously, another new species of the same genus, Candidatus "Scalindua sorokinii", was detected in the water column of the Black Sea, making this genus the most widespread of all anammox bacteria described so far.