厌氧氨氧化菌的种类、特性与检测

厌氧氨氧化是地球氮素循环的重要环节,也是废水生物脱氮和污染环境修复的重要基础;厌氧氨氧化菌作为厌氧氨氧化功能的执行者,近年来成为微生物、环境、地学等领域的研究热点.本文从厌氧氨氧化菌种类、菌种特性及检测手段3个方面综述近年国内外厌氧氨氧化菌研究进展.基于多相分类法,以遗传型分类为主,目前共鉴定厌氧氨氧化菌6属21种,其中Candidatus Anammoximicrobium为最新属.不同种厌氧氨氧化菌在形态结构、细胞组成、生理生化、生态分布存在异同,对温度、盐度、有机物等环境因素的敏感度导致其生态位的差异性,有利于工程应用.近来结合PCR-DGGE、FISH和q PCR等先进的现代分子生物学手段,已形成一套研究厌氧氨氧化菌种类、数量、分布和活性的有效方法,并获得系列基于An AOB 16S r RNA及功能基因(nir S,hzo)的特异性引物.最后提出完善厌氧氨氧化菌纯培物的分离方法、探明厌氧氨氧化菌的代谢途径和遗传特性、明确厌氧氨氧化菌在生境中的分布及贡献是未来研究的热点和难点.     

接种污泥源对厌氧氨氧化启动效能的影响

厌氧氨氧化(anaerobic ammonium oxidation,ANAMMOX)菌生长缓慢是影响其实际应用的主要问题之一,选取合适的接种污泥十分重要.本研究采用好氧污泥、厌氧颗粒污泥和厌氧消化污泥3种接种污泥,分别经过61、70和85 d的运行均实现了厌氧氨氧化过程,氨氮去除率分别为82%、92%和91%,总氮去除率达76%、82%和80%.分别接种3种污泥源的厌氧序批式反应器(ASBR)R1、R2、1t3出水pH值最终稳定在8.4、8.5、8.5.好氧污泥呈絮状,但沉降性比接种前好,厌氧颗粒污泥解体后最终形成粒径集中在0.5～1.0 mm的污泥,厌氧消化污泥则呈沙化状态,有细小颗粒出现.根据厌氧氨氧化细胞产率系数及NH4+-N、NO2--N去除量和NO3--N生成量之间的计量学关系,估算出厌氧氨氧化菌产率系数(以1 mol NH4+产生的CH2O0.5N0.15量计)分别为0.080、0.105和0.114 mol·mol-1,说明反应器内厌氧氨氧化菌有不同程度的衰减.总体而言,厌氧颗粒污泥是富集厌氧氨氧化菌的最适污泥源.     

厌氧氨氧化反应器研究进展

厌氧氨氧化是废水生物脱氮研究的新领域,厌氧氨氧化反应器影响厌氧氨氧化菌的富集、厌氧氨氧化过程的启动、运行的稳定性和处理效果,是其中十分重要的研究内容.本文根据厌氧氨氧化反应的基本特征,分析了反应过程对反应器的主要要求;通过对固定床反应器、流化床反应器、气提式反应器、上流式厌氧污泥床(UASB)等反应器的运行参数和运行结果的比较,分析了各种类型反应器的主要优缺点,并对反应器今后的发展方向提出了建议.表2参37     

金属元素对厌氧氨氧化生化过程和工艺运行的影响

厌氧氨氧化为高效低耗生物脱氮技术,但金属元素对该工艺的作用存在两面性,一方面其促进作用可提高工艺处理效果,另一方面抑制作用使其工程应用受到一定限制.为此本文从生物化学和污水处理工艺的角度,系统地总结了国内外金属对厌氧氨氧化影响的研究现状,重点分析与讨论了不同金属元素对厌氧氨氧化细胞结构和相关酶蛋白活性等的影响和作用机理;综述了金属元素对厌氧氨氧化工艺运行的影响,讨论与分析了不同厌氧氨氧化工艺的半抑制浓度及活性修复方法.     

厌氧氨氧化污水处理工艺及其实际应用研究进展

厌氧氨氧化(Anammox)反应是指在厌氧或者缺氧条件下,厌氧氨氧化微生物以NO2--N为电子受体,氧化NH4+-N为氮气的生物过程。该过程是一种新型自养生物脱氮反应,反应无需外加有机碳源,且污泥产生量小,相对于传统硝化/反硝化脱氮工艺具有显著优势,对处理含高氨氮废水特别是低有机碳源废水具有重大的潜在实际应用价值。近年来,厌氧氨氧化为主体的污水处理工艺已经在各种类型废水处理中得到成功应用,取得了显著的经济和环境效益。综述了厌氧氨氧化反应中常用的亚硝化-厌氧氨氧化工艺(Sharon-Anammox工艺)和完全自养脱氮工艺(CANON工艺)的作用原理、环境调控因子与功能性微生物种群动态分布等最新研究进展,且阐述了两工艺在垃圾渗滤液、厌氧消化液和猪场养殖废水等低碳氮比废水的处理应用效能和最优化控制参数等,为厌氧氨氧化为主体的污水处理工艺的工程化应用提供了技术支撑。最后,总结并介绍了国内外厌氧氨氧化工艺现场规模化应用实例和控制参数,同时,对厌氧氨氧化污水处理工艺实际应用的研究前景及亟待解决的问题进行了展望,认为现场应用中Anammox菌的快速富集培养、有机碳源对Anammox菌的抑制效应以及厌氧氨氧化工艺的广谱适用性等将是厌氧氨氧化工艺大规模应用的难点和热点问题。为厌氧氨氧化工艺实际应用控制和推广提供了理论基础,具有重要的理论和现实意义。     

硝化作用微生物的分子生物学研究进展

回顾了硝化作用微生物的分子生物学研究进展 ,主要介绍了硝化作用微生物的种类 ,包括氨氧化菌、亚硝酸氧化菌、异养氨氧化菌和厌氧氨氧化菌 .这些微生物的系统发育分析表明亚硝化菌的系统发育相对简单 ,而硝化细菌则要复杂许多 .还介绍了在硝化作用微生物生态学研究中应用的分子生物学技术 ,如PCR、变性梯度胶电泳 (DGGE)、原位荧光杂交 (FISH)等 ,以及不同类群细菌中与硝化作用相关的酶类及其基因 .文章的最后还提出了一些研究展望 .图 3参 35     

碳氮比对晚期垃圾渗滤液自养脱氮的影响

采用缺氧/好氧(A/O)短程硝化与升流式厌氧污泥床(UASB)厌氧氨氧化组合的自养脱氮系统处理实际晚期渗滤液,重点考察了碳氮比(C/N)对该系统的综合影响.在较低C/N范围内(1.45到1.95),氨氮去除率变化不显著,当C/N达到2.46以上时,硝化和厌氧氨氧化活性恶化导致其去除率从97.7%降为83.3%.化学需氧量和总氮去除率整体随C/N的提高而呈现先增加后减少的趋势.适度反硝化补充的碱度不仅为硝化菌提供充足的无机碳源,且有利于保障较高的p H和游离氨以维持稳定的短程硝化,亚硝积累率随C/N的提升呈现小幅上升.种群结构分析表明高C/N与厌氧氨氧化菌群所占的比例呈反比关系,从1.65%降为0.31%,维持厌氧氨氧化菌和反硝化菌的动态平衡是保证系统正常运行的关键因素.C/N为1.95时系统整体性能最优.     

铁氨氧化污水脱氮的应用研究进展

铁氨氧化(Feammox)是一种以廉价、易得的铁作为微生物电子供体的新型自养生物脱氮技术，即Fe(Ⅲ)还原与厌氧氨氧化的结合工艺，拥有成本低廉、无需有机碳源、污泥产量小、无温室气体产生等优势，是污水处理的一种潜在脱氮途径.本文对铁氨氧化反应的机理、功能菌种的种类和特性及电子穿梭体对其的影响进行了介绍，总结了铁氨氧化在污水环境中的脱氮效果及其与厌氧氨氧化、硝酸盐依赖型亚铁氧化和生物电化学系统的耦合技术，并指出目前铁氨氧化的应用问题及该技术未来的研究方向和重点可能是菌分离纯化、工艺参数控制.     

太湖梅梁湾沉积物中铁氧化氨作用及其脱氮贡献

铁氧化氨反应是最近发现的一种新型氮转化途径,在林地、水稻田和湿地土壤氮循环过程中具有重要作用。然而,鲜有研究关注富营养化湖泊沉积物中的铁氧化氨过程。该研究在调查太湖梅梁湾沉积物理化性质和主要铁还原菌丰度的基础上,采用同位素示踪技术和C_2H_2抑制法研究了沉积物的铁还原速率和铁氧化氨过程,以实验过程中~(30)N_2和~(29)N_2的产生速率核算了沉积物的铁氧化氨速率。通过考察沉积物相关理化性质、铁还原菌丰度与铁氧化氨速率之间相关性,确定了这些因子对铁氧化氨的影响。结果表明:在太湖梅梁湾4个采样点的沉积物中均存在铁氧化氨过程,该过程能够在厌氧条件下将NH_4~+直接氧化为N_2,或者将NH_4~+氧化为NO_2~-、NO_3~-,然后厌氧氨氧化或反硝化过程将NO_2~-、NO_3~-转化为N_2导致沉积物氮损失。梅梁湾沉积物铁氧化氨速率范围为0.28~0.43 kg~(-1)·d~(-1),占太湖人为输入无机氮的6.1%~9.4%。沉积物Fe(Ⅲ)含量和TOC含量与铁氧化氨速率之间呈显著相关性(P0.05),在铁氧化氨过程中起重要作用。相反,pH与铁氧化氨之间无显著相关性(P0.05)。地杆菌属(Geobacteraceae spp.)、希瓦氏茵属(Shewanella spp.)、酸微菌科(Acidimicrobiaceae)及微酸菌A6属(Acidimicrobiaceae A6)与铁氧化氨呈显著相关性(P0.05),表明铁还原菌可能直接参与铁氧化氨过程。综上,铁氧化氨是富营养化湖泊沉积物中氮素迁移转化的重要途径之一。     

废水生物脱氮中N_2O和NO_x的产生和作用

废水生物脱氮中N2O和NOx来源于硝化、反硝化、厌氧氨氧化和化学反硝化等过程.电子受体和供体浓度、pH、缓冲剂类型、有机负荷、微生物种类及其相互作用等都会影响这些气态中间产物的产生.NO2能够氧化氨和强化好氧和厌氧氨氧化,NO能够阻止C2H2对好氧氨氧化活性的抑制,两者对好氧氨氧化活性的恢复至关重要.所有这些表明,废水生物脱氮的气态中间产物N2O和NOx在氮的生物转化中具有重要的正面作用,甚至必不可少.基于NO2曝气技术和Brocadiaanammoxidans与Nitrosomonas协同作用的废水生物脱氮新技术开发是今后一段时间的重要研究方向.图4参35     

氮肥对土壤氧化大气甲烷影响的机制

综合评述了氮素对土壤氧化甲烷的抑制机制。包括 :( 1)竞争甲烷单氧化酶的竞争抑制机制 ,( 2 )代谢产物的毒害抑制机制 ,( 3)外源盐引起的微生物生理缺水抑制机制和 ( 4)氮素周转作用引起的抑制机制。提出了氧化菌竞争利用土壤空气有限O2 的竞争抑制机制 ,即氨氧化菌利用更多的土壤有限氧气→产生优势氨氧化菌→形成优势菌群→限制甲烷氧化菌繁殖和功能发挥的氨长期抑制土壤氧化大气甲烷的机制 ,并认为这种抑制作用是不可逆的     

厌氧除磷同步脱氮及影响因素研究

采用鸡粪污泥为种泥,在厌氧混合连续流反应装置内进行厌氧还原磷产生磷化氧功能菌的富集,进行硝酸盐、硫酸盐、不同碳源和氮源条件下厌氧除磷效率的研究,并考察磷化氧的生成与硝酸、总磷、氨氮去除的关系.结果表明,(1)SO_4~(2-)-S适宜的投加量为26 mg·L~(-1),不投加NO_3~--N.水中含有氧化态的无机物在厌氧条件下与磷争夺[H]导致厌氧除磷的效率下降.(2)合适的碳源为葡萄糖1 000 mg·L~(-1),纤维素不适合作为碳源,合适的氮源为蛋白胨500 mg·L~(-1),水中含有的还原糖和有机氮源促进磷化氧的生成.(3)pH值控制在6.5～7.0的范围,最适宜的生长温度在35℃左右.(4)氨氮的去除率随着总磷的去除率而增加,在厌氧条件下可达到同时脱氮除磷的效果.磷的去除由厌氧除磷菌还原磷生成磷化氧完成,氨氮由生成氮气或生成蛋白质来去除.     

Biological conversion pathways of sulfate reduction ammonium oxidation in anammox consortia

• The SRAO phenomena tended to occur only under certain conditions. • High amount of biomass and non-anaerobic condition is requirement for SRAO. • Anammox bacteria cannot oxidize ammonium with sulfate as electron acceptor. • AOB and AnAOB are mainly responsible for ammonium conversion. • Heterotrophic sulfate reduction mainly contributed to sulfate conversion. For over two decades, sulfate reduction with ammonium oxidation (SRAO) had been reported from laboratory experiments. SRAO was considered an autotrophic process mediated by anammox bacteria, in which ammonium as electron donor was oxidized by the electron acceptor sulfate. This process had been attributed to observed transformations of nitrogenous and sulfurous compounds in natural environments. Results obtained differed largely for the conversion mole ratios (ammonium/sulfate), and even the intermediate and final products of sulfate reduction. Thus, the hypothesis of biological conversion pathways of ammonium and sulfate in anammox consortia is implausible. In this study, continuous reactor experiments (with working volume of 3.8L) and batch tests were conducted under normal anaerobic (0.2≤DO<0.5 mg/L) / strict anaerobic (DO<0.2 mg/L) conditions with different biomass proportions to verify the SRAO phenomena and identify possible pathways behind substrate conversion. Key findings were that SRAO occurred only in cases of high amounts of inoculant biomass under normal anaerobic condition, while absent under strict anaerobic conditions for same anammox consortia. Mass balance and stoichiometry were checked based on experimental results and the thermodynamics proposed by previous studies were critically discussed. Thus anammox bacteria do not possess the ability to oxidize ammonium with sulfate as electron acceptor and the assumed SRAO could, in fact, be a combination of aerobic ammonium oxidation, anammox and heterotrophic sulfate reduction processes.     

Exponential Collapse of Benthic Depurative Capacity in A Eutrophic Tropical Marine Ecosystem: A Threshold Response to Organic Nitrogen Loading

Microbial decomposition of particulate organic matter in sediments can substantially modulate eutrophication of nearshore marine environments. Flux budgets for nitrogen compounds and quantification of rates of microbial transformation can provide important information on the process of eutrophication. This survey documents sediment nitrogen budgets for a eutrophic nearshore marine environment at La Parguera on southwest coast of Puerto Rico, including consideration of the organic fraction in addition to microbial transformations.

Sediments of the inshore channel at La Parguera denote the input of heavy organic loads with low redox potentials and high ammonium contents. Benthic fluxes of dissolved organic nitrogen are below those recorded for ammonium. These observations suggest that nearshore tropical sediments subject to heavy nitrogen loads act as a sink for organic nitrogen and that ammonification is a key process in the release of nitrogen from sediments to the water column. Nitrification and denitrification rates are low overall and inversely related to the redox potential. Depuration of excess nitrogen through denitrification is ineffective in these sediments. in contrast to more robust temperate environments, our work demonstrates that tropical marine systems are particularly susceptible to eutrophication given their limited capacity for depuration of excess nitrogen.     

Exponential Collapse of Benthic Depurative Capacity in A Eutrophic Tropical Marine Ecosystem: A Threshold Response to Organic Nitrogen Loading

Microbial decomposition of particulate organic matter in sediments can substantially modulate eutrophication of nearshore marine environments. Flux budgets for nitrogen compounds and quantification of rates of microbial transformation can provide important information on the process of eutrophication. This survey documents sediment nitrogen budgets for a eutrophic nearshore marine environment at La Parguera on southwest coast of Puerto Rico, including consideration of the organic fraction in addition to microbial transformations.

Sediments of the inshore channel at La Parguera denote the input of heavy organic loads with low redox potentials and high ammonium contents. Benthic fluxes of dissolved organic nitrogen are below those recorded for ammonium. These observations suggest that nearshore tropical sediments subject to heavy nitrogen loads act as a sink for organic nitrogen and that ammonification is a key process in the release of nitrogen from sediments to the water column. Nitrification and denitrification rates are low overall and inversely related to the redox potential. Depuration of excess nitrogen through denitrification is ineffective in these sediments. in contrast to more robust temperate environments, our work demonstrates that tropical marine systems are particularly susceptible to eutrophication given their limited capacity for depuration of excess nitrogen.     

不同氮源对4种海洋微藻生长的影响

采用实验室一次性培养的方法,研究了硝氮、氨氮、尿素和混合氨基酸等4种不同氮源对典型赤潮藻赤潮异弯藻Heterosigma akashiwo、凯伦藻Karenia sp.、球形棕囊藻Phaeocystis globosa和常见浮游植物优势种类角毛藻Chaetoceros sp.生长的影响。结果表明,这4种海洋微藻不仅能利用无机氮硝氮和氨氮,而且也均能利用有机氮尿素和混合氨基酸。赤潮异弯藻、凯伦藻和角毛藻均在以硝氮为唯一氮源时,比生长速率分别达到最大值0.45、0.52和0.70 d-1;而球形棕囊藻在以硝氮和尿素为唯一氮源时,比生长速率均达最大值0.65 d-1。可溶性有机氮库中的重要组成成分尿素和氨基酸均能显著促进4种海洋微藻的生长;相比较而言,赤潮异弯藻和凯伦藻更加喜好有机氮氨基酸,而球形棕囊藻和角毛藻更加喜好尿素。海洋微藻具备利用有机氮源的能力,无疑扩展了其氮营养来源,在无机氮缺乏而有机氮丰富的水体中,它们在浮游植物群落中更具有竞争优势。     

氮肥对旱地土壤甲烷氧化能力的影响

甲烷是重要的温室气体之一，对全球变暖的贡献率是20％。大气中甲烷浓度的迅速增加与甲烷的源和汇的不平衡有关。土壤中存在甲烷氧化菌，在一定条件下可以氧化大气中的甲烷。不过土壤对甲烷的氧化能力受一些农业生产活动的影响。氮肥的施用对土壤甲烷氧化有一定的影响。长期定位施肥实验的结果表明氮肥对土壤甲烷氧化的影响主要来源于铵态氮而不是硝态氮，因为氨对甲烷氧化有着竞争性抑制作用。此外，长期施用氮肥还改变了土壤微生物的区系及其活性，从而降低甲烷的氧化速率。氮肥施用的短期效应则与土壤的耕作管理历史、土壤性质、氮肥施用量等因素有关。有机肥对土壤甲烷氧化的影响比化肥要低得多，其物理化学性质及施用方法的不同都会影响到土壤对甲烷的氧化。     

Mechanism studies on nitrogen removal when treating ammonium-rich leachate by sequencing batch biofilm reactor

The nitrogen removal mechanism was studied and analyzed when treating the ammonium-rich landfill leachate by a set of sequencing batch biofilm reactors (SBBRs), which was designed independently. At the liquid temperature of (32 ± 0.4)°C, and after a 58-days domestication period and a 33-days stabilization period, the efficiency of ammonium removal in the SBBR went up to 95%. Highly frequent intermittent aeration suppressed the activity of nitratebacteria, and also eliminated the influence on the activity of anaerobic ammonium oxidation (ANAMMOX) bacteria and nitritebacteria. This influence was caused by the accumulation of nitrous acid and the undulation of pH. During the aeration stage, the concentration of dissolved oxygen was controlled at 1.2–1.4 mg/L. The nitritebacteria became dominant and nitrite accumulated gradually. During the anoxic stage, along with the concentration debasement of the dissolved oxygen, ANAMMOX bacteria became dominant; then, the nitrite that was accumulated in the aeration stage was wiped off with ammonium simultaneously.     

新型废水生物脱氮的微生物学研究进展

生物脱氮是含氮废水处理公认的最佳处理方式,随着对生物脱氮微生物学原理研究的不断深入,许多新的生物脱氮特殊菌株或菌群及微生物转化机制不断被发现.本文在传统生物脱氮过程机理上,结合最近国内外生物脱氮的新发现,就短程硝化反硝化、同时硝化反硝化、厌氧氨氧化的微生物学原理进行了阐述.图1表2参23     

Simultaneous Feammox and anammox process facilitated by activated carbon as an electron shuttle for autotrophic biological nitrogen removal

• The autotrophic nitrogen removal combining Feammox and Anammox was achieved. • Activated carbon can be used as an electron shuttle to enhance Feammox activity. • Fe(III) was reduced to Fe(II) and the secondary Fe(II) mineral (FeOOH) was obtained. • The iron-reducing bacteria and Anammox consortium was enriched simultaneously. Ferric iron reduction coupled with anaerobic ammonium oxidation (Feammox) is a novel ferric-dependent autotrophic process for biological nitrogen removal (BNR) that has attracted increasing attention due to its low organic carbon requirement. However, extracellular electron transfer limits the nitrogen transformation rate. In this study, activated carbon (AC) was used as an electron shuttle and added into an integrated autotrophic BNR system consisting of Feammox and anammox processes. The nitrogen removal performance, nitrogen transformation pathways and microbial communities were investigated during 194 days of operation. During the stable operational period (days 126–194), the total nitrogen (TN) removal efficiency reached 82.9%±6.8% with a nitrogen removal rate of 0.46±0.04 kg-TN/m³/d. The contributions of the Feammox, anammox and heterotrophic denitrification pathways to TN loss accounted for 7.5%, 89.5% and 3.0%, respectively. Batch experiments showed that AC was more effective in accelerating the Feammox rate than the anammox rate. X-ray photoelectron spectroscopy (XPS) analyses showed the presence of ferric iron (Fe(III)) and ferrous iron (Fe(II)) in secondary minerals. X-ray diffraction (XRD) patterns indicated that secondary iron species were formed on the surface of iron-AC carrier (Fe/AC), and Fe(III) was primarily reduced by ammonium in the Feammox process. The phyla Anaerolineaceae (0.542%) and Candidatus Magasanikbacteria (0.147%) might contribute to the Feammox process, and Candidatus Jettenia (2.10%) and Candidatus Brocadia (1.18%) were the dominative anammox phyla in the bioreactor. Overall, the addition of AC provided an effective way to enhance the autotrophic BNR process by integrating Feammox and anammox.