铁矿物强化厌氧氨氧化效能及其微生物机制研究进展

自然界中的氮循环与铁循环相互交联,参与氮循环的厌氧氨氧化（anaerobic ammonium oxidation,anammox）菌的生长代谢及活性发挥也与铁元素紧密关联。自然界广泛存在的铁矿物因具有运行成本低廉、稳定性好、二次污染小等优势,在污水处理领域得到广泛应用。在厌氧氨氧化脱氮系统中引入适量铁矿物,不仅有助于促进anammox菌和铁还原菌的富集,提高功能基因丰度和相关酶活性,还可能通过影响污泥浓度、血红素c含量、胞外聚合物含量和颗粒化程度,改善污泥性能和提高厌氧氨氧化系统的稳定性。同时,铁矿物具有促进体系多种氮素转化途径（如anammox、铁自养反硝化、铁氨氧化、异化硝酸盐还原成铵和反硝化）相耦合的潜能,可以提高anammox污水处理系统的总氮去除率。本文基于铁矿物在促进污水生物脱氮方面的良好性能及其在anammox系统中的变化,从脱氮效能、污泥特性、微生物特征及酶活性等方面,系统综述了铁矿物对厌氧氨氧化系统的强化作用机制,并从anammox菌对铁矿物的利用及铁元素的摄取角度展望了后续的研究方向,以期为铁矿物强化厌氧氨氧化系统的实际应用提供理论和技术指导。     

湖泊微生物反硝化过程及速率研究进展

湖泊中微生物介导的反硝化过程对于区域乃至全球的气候环境变化有着深远的影响。因此,研究湖泊微生物反硝化过程及速率有助于我们深刻理解湖泊氮元素生物地球化学循环规律,全面认识湖泊生境对全球氮循环的贡献。本文综述了湖泊生境中反硝化过程(包括典型的反硝化过程及与其他物质循环耦合的反硝化过程,如与有机氮耦合的共反硝化作用、与碳循环耦合的硝酸盐/亚硝酸盐依赖型厌氧甲烷氧化、与铁循环耦合的硝酸盐依赖型铁氧化、与硫循环耦合的硝酸盐还原硫氧化)的速率、驱动微生物及其影响因素。最后对湖泊反硝化过程研究现状和未来发展方向提出总结与展望。     

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。     

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.     

亚硝酸盐型甲烷厌氧氧化及其功能微生物研究进展

亚硝酸盐型甲烷厌氧氧化（nitrite-dependent anaerobic methane oxidation,N-DAMO）是耦合氮循环和碳循环的关键环节,主要是由亚硝酸盐型甲烷厌氧氧化菌（Candidatus Methylomirabilis oxyfera）介导完成,对于研究全球氮和碳元素的生物地球化学循环具有重要意义。本文首先总结了国内外N-DAMO的影响因素和在不同自然生态系统中的分布;然后阐述了N-DAMO菌的生理生化特性及其富集培养优化实验和检测技术,最后探讨了N-DAMO技术的应用现状。本综述不仅有助于揭示全球碳氮循环的耦合作用机制,也为N-DAMO反应耦合其他厌氧生物处理过程应用到污水的除碳脱氮上提供了理论依据。     

New concepts of microbial treatment processes for the nitrogen removal in wastewater

Many countries strive to reduce the emissions of nitrogen compounds (ammonia, nitrate, NOx) to the surface waters and the atmosphere. Since mainstream domestic wastewater treatment systems are usually already overloaded with ammonia, a dedicated nitrogen removal from concentrated secondary or industrial wastewaters is often more cost-effective than the disposal of such wastes to domestic wastewater treatment. The cost-effectiveness of separate treatment has increased dramatically in the past few years, since several processes for the biological removal of ammonia from concentrated waste streams have become available. Here, we review those processes that make use of new concepts in microbiology: partial nitrification, nitrifier denitrification and anaerobic ammonia oxidation (the anammox process). These processes target the removal of ammonia from gases, and ammonium-bicarbonate from concentrated wastewaters (i.e. sludge liquor and landfill leachate). The review addresses the microbiology, its consequences for their application, the current status regarding application, and the future developments.     

Nitrogen removal from animal waste treatment water by anammox enrichment

The aim of this work was to examine the applicability of the anaerobic ammonium oxidation (anammox) process to three kinds of low BOD/N ratio wastewaters from animal waste treatment processes in batch mode. A rapid decrease of NO(2)(-) and NH(4)(+) was observed during incubation with wastewaters from AS and UASB/trickling filter and their corresponding control artificial wastewaters. This nitrogen removal resulted from the anammox reaction, because the ratio of removed NO(2)(-) and NH(4)(+) was close to the theoretical ratio of the anammox reaction. Comparison of the inorganic nitrogen removal rate of the actual wastewater and that of control artificial wastewater showed that these two kinds of wastewater were very suitable for anammox treatment. Incubation with wastewater from RW did not show a clear anammox reaction; however, diluting it by half enabled the reaction, suggesting the presence of an inhibitory factor. This study showed that the three kinds of wastewater from animal waste treatment processes were suitable for anammox treatment.     

极端酸性矿山环境微生物基于CRISPR系统的适应性免疫机制

【目的】通过对酸性矿山环境中嗜酸硫杆菌属(Acidithiobacillus)、脱硫弧菌属(Desulfovibrio)、钩端螺旋菌属(Leptospirillum)、硫化杆菌属(Sulfobacillus)、酸原体属(Acidiplasma)和铁质菌属(Ferroplasma)的100株冶金微生物基因组中CRISPR-Cas系统的结构特征和同源关系进行生物信息学分析,在基因组水平上解析冶金微生物基于CRISPR系统对极端环境的适应性免疫机制。【方法】从NCBI网站下载基因组序列,采用CRISPR Finder定位基因组中潜在的CRISPR簇。分析CRISPR系统的组成结构与功能:利用Clustal Omega对重复序列(repeat)分类;将间隔序列(spacer)分别与nr数据库、质粒数据库和病毒数据库比对,获得注释信息;根据Cas蛋白的种类和同源性对酸性矿山环境微生物的CRISPR-Cas系统分型。【结果】在100株冶金微生物基因组中共鉴定出415个CRISPR簇,在176个c CRISPR簇中共有80种不同的重复序列和4147条间隔序列。对重复序列分类,发现12类重复序列均能形成典型的RNA二级结构,Cluster10中的重复序列在冶金微生物中最具有代表性。间隔序列注释结果表明,这些微生物曾遭受来自细菌质粒与病毒的攻击,并通过不同的防御机制抵抗外源核酸序列的入侵。冶金微生物细菌的大部分CRISPR-Cas系统属于I-C和I-E亚类型,而古菌的CRISPR-Cas系统多为I-D亚类型,两者基于CRISPR-Cas系统的进化过程中存在显著差异。【结论】酸性矿山环境微生物的CRISPR结构可能采用不同免疫机制介导外源核酸序列与Cas蛋白的相互作用,为进一步揭示极端环境微生物的适应性进化机理奠定了基础。     

硝态氮还原型厌氧甲烷氧化的研究进展及展望

以硝态氮为电子受体的甲烷厌氧氧化(Nx-damo)是近年被证实的微生物驱动的地球氮、碳循环机制,对于认识重要元素地球化学循环的微生物驱动机制和自然环境中甲烷的源与汇具有重大意义;在废水生物脱氮及其温室气体减排方面也具有潜在工程应用价值。从功能微生物富集及其影响因素、生理特性、生物代谢的可能机理等方面对Nx-damo的最新进展进行了梳理和讨论;评估了其应用于废水处理的潜力和优势;对未来的研究方向进行了展望,以期推动该领域更广泛的研究,并为其提供有价值的参考。     

微生物甲烷氧化反硝化耦合反应研究进展

甲烷氧化反硝化耦合过程是连接碳循环和氮循环的重要桥梁.该过程的深入研究有助于完善人们对全球碳氮生物化学循环的认识.甲烷作为反硝化外加气体碳源,既能调控大气甲烷平衡,有效减缓由甲烷引起的温室效应,又能降低反硝化工艺中因投入外加碳源带来的成本.因此近年来甲烷氧化反硝化耦合反应及其机理研究倍受关注.本文主要讨论了好氧和厌氧两种类型的甲烷氧化反硝化过程,重点对其微生物耦合反应机理及其影响因素进行了综述,同时指出了其工程化应用存在的问题,并对其应用前景提出展望.
     

有机碳源下废水厌氧氨氧化同步脱氮除碳

为明确有机碳源胁迫下,厌氧氨氧化反应器的同步脱氮除碳规律及功能微生物群落结构的动态变化,采用成功启动的厌氧氨氧化UASB反应器,通过逐步提升进水有机负荷,探究有机碳源下废水厌氧氨氧化同步脱氮除碳。研究表明,当进水化学需氧量(Chemical oxygen demand,COD)浓度从172 mg/L升至620 mg/L,反应器维持较高的脱氮效率,氨氮和总氮去除率均在85%以上,并对COD具有平均56.6%的去除率,高浓度COD未对Anammox菌活性构成显著抑制作用。聚合酶链式反应和变性梯度凝胶电泳(PCR-DGGE)图谱和割胶测序结果表明,变形菌门Proteobacteria、浮霉菌门Planctomycetes、绿曲挠菌门Chloroflexi以及绿菌门Chlorobi等微生物共存于同一反应体系中,推测反应器内存在复杂的脱氮除碳途径。而且,代表厌氧氨氧化的部分浮霉菌门微生物能耐受高浓度有机碳源,在高有机负荷下依旧发挥着高效的脱氮作用,为反应器高效脱氮提供了保障。     

厌氧氨氧化菌脱氮机理及其在污水处理中的应用

厌氧氨氧化细菌(anammox)可以将亚硝酸盐和氨氮转化为氮气从而缩短氨氮转化的过程,它已经成为新型生物污水脱氮技术研究的热点之一。当前,有关厌氧氨氧化菌特有的生理结构特点、种群分类及其功能酶等方面的研究取得了一定突破,为实现其工业应用奠定了良好的理论基础;同时分子生物学技术在厌氧氨氧化细菌种群分布、群落多样性及其共生关系等方面的应用也大大促进了污水生物脱氮技术的革新和进步。总结了厌氧氨氧化菌主要的生理生化特点、细胞结构特点、脱氮机理、污水处理体系中的应用以及分子生物学方法对污水处理体系中厌氧氨氧化菌种群分析的研究现状,并指出未来anammox细菌在生物特性及在污水脱氮处理实际应用的研究中的热点问题。生物特性方面的主要研究热点有：（1）anammox细菌除厌氧氨氧化作用外,其它新陈代谢途径有待探索;（2）anammox细菌在不同环境中分布的倾向性问题;（3）新型anammox细菌的确定。污水处理的实际应用方面的主要研究热点有：（1）anammox污泥的快速高效富集问题;（2）设计高特异性引物;（3）anammox细菌和其他微生物的共生关系。     

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.     

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.     

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。     

Patterns and drivers of anaerobic nitrogen transformations in sediments of thermokarst lakes

Significant attention has been given to the way in which the soil nitrogen (N) cycle responds to permafrost thaw in recent years, yet little is known about anaerobic N transformations in thermokarst lakes, which account for more than one-third of thermokarst landforms across permafrost regions. Based on the N isotope dilution and tracing technique, combined with qPCR and high-throughput sequencing, we presented large-scale measurements of anaerobic N transformations of sediments across 30 thermokarst lakes over the Tibetan alpine permafrost region. Our results showed that gross N mineralization, ammonium immobilization, and dissimilatory nitrate reduction rates in thermokarst lakes were higher in the eastern part of our study area than in the west. Denitrification dominated in the dissimilatory nitrate reduction processes, being two and one orders of magnitude higher than anaerobic ammonium oxidation (anammox) and dissimilatory nitrate reduction to ammonium (DNRA), respectively. The abundances of the dissimilatory nitrate reduction genes (nirK, nirS, hzsB, and nrfA) exhibited patterns consistent with sediment N transformation rates, while α diversity did not. The inter-lake variability in gross N mineralization and ammonium immobilization was dominantly driven by microbial biomass, while the variability in anammox and DNRA was driven by substrate supply and organic carbon content, respectively. Denitrification was jointly affected by nirS abundance and organic carbon content. Overall, the patterns and drivers of anaerobic N transformation rates detected in this study provide a new perspective on potential N release, retention, and removal upon the formation and development of thermokarst lakes.     

Reactivation of biocatalysts after long-term storage and startup of the DEAMOX process

Studies of our laboratory have led to elaboration of the DEAMOX (DEnitrifying AMmonia OXidation) technology intended for removal of nitrogen contaminants from wastewater. The DEAMOX process comprises two anaerobic stages implemented by the same sludge biocatalyst, namely, denitratation (conversion of nitrate to nitrite) and anammox reaction (ANaerobic AMmonium nitrogen OXidation by nitrite). The results of reactivation of biocatalysts after their long-term storage (5 and 16 months) and successful startup of the DEAMOX process in two modifications (S- and O-) are described. An S-DEAMOX process was launched using a sludge biocatalyst with restored anammox activity of 20.1 mg N/g VSS/day; this process provided removal of 78% of nitrogen in reactor over 20 days. The launched O-DEAMOX process with the sludge biocatalyst with anammox activity of 6.1 mg N/g VSS/day provided for 87% removal of the total nitrogen compounds over 30 days. Two different electron donors were used at the stage of nitrate conversion to nitrite, namely, an inorganic donor, sulfide (S-DEAMOX), and an organic one, acetate (O-DEAMOX).     

自然生境中亚硝酸盐型厌氧甲烷氧化细菌研究进展

随着功能微生物介导的亚硝酸盐型厌氧甲烷氧化(nitrite-dependent anaerobic methane oxidation,N-DAMO)过程被发现,人们对自然界的碳氮循环有了全新的认识,该过程成为自然生态系统中温室气体甲烷的汇,同时还是氮污染的消减途径。本文系统介绍了N-DAMO过程反应机理以及参与该过程的亚硝酸盐型厌氧甲烷氧化细菌(Candidatus Methylomirabilis oxyfera)的生理生化特征,并对研究该功能菌的分子微生物方法进行了汇总。通过对不同自然生境中该细菌的研究报道进行总结分析,揭示各生境中年均降水量、年均温度、所处不同自然区等大尺度宏观环境因子及碳源、氮源、pH和氧气含量等生存因子对其群落结构的潜在影响,最后在展望中提出此功能菌在未来可深入研究的方向,期望能厘清厌氧甲烷氧化过程及其功能菌在碳、氮循环中的生态学功能。     

双阳极室甲烷微生物燃料电池同步脱氮除硫性能及微生物群落分析

【背景】甲烷厌氧氧化(anaerobic oxidation of methane, AOM)包含反硝化型甲烷厌氧氧化和硫酸盐还原型甲烷厌氧氧化。目前,人们向水体中排放过量的含氮及含硫污染物,引起了严重的环境污染和生态破坏。【目的】利用甲烷厌氧氧化微生物燃料电池(microbial fuel cell, MFC)研究同步脱氮除硫耦合反应机理及反应过程中微生物的多样性信息。【方法】构建了3个微生物燃料电池(N-S-MFC、N-MFC、S-MFC),以甲烷作为唯一碳源,探究其同步脱氮除硫性能,并采用16S rRNA基因高通量测序技术对微生物群落结构进行分析。【结果】N-S-MFC中硝酸盐和硫酸盐的去除率分别为90.91%和18.46%。阳极室中微生物的相对丰度提高,与反硝化及硫酸盐还原菌相关的微生物大量富集,如门水平上拟杆菌门(Bacteroidota)、厚壁菌门(Firmicutes)和脱硫杆菌门(Desulfobacterota),同时属水平上Methylobacterium＿Methylorubrum、Methylocaldum、Methylomonas等常见的甲烷氧化菌增多。【结论...