全程自养脱氮反应系统的微生物区系分析*

在建立全程自养脱氮反应器的基础上,以活性污泥为对照,分析了脱氮反应器内真菌、细菌和放线菌的数量、种类(类群)、种(株系)数和优势种(株系或类群),及硝化菌和亚硝化菌的数量变化。研究结果表明,与活性污泥相比,全程自养脱氮反应器内微生物数量、种类和区系组成发生很大变化。自养脱氮反应器内亚硝化菌数量显著增加,说明亚硝化菌的积累是全程自养脱氮系统的一个显著特点。     

单级自养脱氮系统亚硝化菌株的分离、鉴定及定性

[目的]研究单级自养脱氮系统中亚硝化菌株的培养及代谢特征,为系统运行操控提出理论指导.[方法]从单级白养脱氮系统活性污泥中采集微生物样品,经过4次富集和分离过程,最终获得一株亚硝化能力较强的菌株Nl,通过显微镜观察及16S rDNA序列分析鉴定该菌株,研究摇瓶装量、pH、温度及底物浓度对其代谢过程的影响.[结果]该菌株与Nitrosomonas sp.NL7(AY958677)、Nitrosomonas AS1(EF016119)、Nitrosomonas sp.Is32(AJ621027)相似性分别为97%、96%和96%,对氧气需求量存在较严格要求,pH及温度对其氨氧化活性具有明显的影响,最适条件分别为pH8.0和30℃,在氨氮浓度80-800 mg/L较宽的底物浓度范围内具有活性,当氨氮浓度高达800 mg/L时,其氨氧化活性没有受到明显的抑制.[结论]该N1菌株为亚硝化单胞菌(Nitrosomonas sp.),与已有报道的其它亚硝化菌相比,N1对氨氮有较强降解能力及较广的浓度适应范围.     

限氧自养硝化-反硝化生物脱氮新技术

限氧自养硝化—反硝化是部分硝化与厌氧氨氧化相耦联的生物脱氮反应过程,通过严格控制溶解氧在0．1～0．3mg·L^-1,实现硝化反应控制在亚硝酸阶段,然后以硝化阶段剩余的NH4^+作为电子供体,在厌氧条件下实现反硝化,该反应过程是完全的自养硝化—反硝化过程,具有能耗低、脱氮效率高、反应系统占地面积小等优点,适用于处理COD／NH4^+—N低的废水,是一种非常有应用前景的生物脱氮技术,文中详细介绍了限氧自养硝化—反硝化生物脱氮反应过程的研究进展,讨论了其微生物学机理及应用前景。     

废水自养生物脱氮技术研究进展

基于短程硝化和厌氧氨氧化的自养脱氮工艺是生物脱氮领域研究的热点,它的发现为低碳氮比废水的处理提供了新的思路。近些年来,人们陆续开发了SHARON、ANAMMOX、CANON、OLAND等自养生物脱氮工艺,进一步推动了高效、低耗脱氮技术的开发和研究。本文从工艺原理、特点等方面,对自养生物脱氮工艺的国内外研究状况进行了总结和对比,并提出了存在的问题及发展方向。     

基于氨单加氧酶基因的自养脱氮菌群结构分析

全程自养脱氮是一种在高氨氮低溶氧条件下完全由自养菌群作用脱除氮素的现象.以全程自养脱氮污泥为研究对象,特异性扩增氨单加氧酶活性基因amoA片段,建立克隆文库并对克隆序列进行系统发育学分析,考察全程自养脱氮系统从建立到退化过程中氨氧化菌的结构变迁.结果表明:Nitrosomonas oligotropha和Nitrosomonas europaea细菌是系统中的主要氨氧化菌,而随着系统的退化前者逐渐被后者完全取代,而氨氧化菌的种群变迁可能并不是全混流系统全程自养脱氮效率下降的原因.     

单级自养脱氮系统中厌氧氨氧化菌的分子生物学鉴定

对具有厌氧氨氧化作用的细菌进行更深入的分析有助于了解该菌在生物脱氮过程的应用。对稳定运行、氨氮转化率及总氮去除率分别达到90%及80%左右的单级自养脱氮系统的底部取活性污泥,采用分子生物学方法提取活性污泥细菌总DNA,利用特异引物Pla46rc/Amx820对单级自养脱氮系统中的厌氧氨氧化菌16S rDNA基因进行PCR扩增。扩增产物经克隆、测序及BLAST分析,结果表明该单级自养脱氮系统中存在的厌氧氨氧化菌与Candidatus Kueneniastuttgartiensis和Candidatus Brocadia anammoxidans的16S rDNA序列同源性达99%,进化分析证明与Candidatus Kuenenia stuttgartiensis进化上较为接近。     

脱氮微生物及脱氮工艺研究进展

脱氮是大部分污水处理系统中不可缺少的一环。由于具有经济高效、工艺简单和无二次污染等显著优势,生物脱氮工艺在最近数十年中备受关注。根据脱氮微生物的生理特性和脱氮机制不同,文中分类综述了近年来生物脱氮工艺的研究进展,重点对比分析了硝化菌、反硝化菌和厌氧氨氧化菌以及以这些菌为基础的不同生物脱氮工艺的优缺点,为复杂污水环境的脱氮工艺选择提供参考。基于微生物脱氮机制,通过合成生物学技术开发高效脱氮菌株,结合不同工艺优点并应用自动化模拟最佳条件,从而建立经济高效的脱氮工艺将是未来发展的重要方向。     

南极长城站微生物区系调查

1992～1993年的夏季,对南极中国长城站的陆地进行了微生物区系的现场调查。先后采集各种类型上样多个。其中对有生态意义的25个样进行了不同类型微生物的分离和计数;中温培养下每克湿土壤菌量为10²～10⁶,用马丁培养基对真菌进行分离培养和计数,每克湿土含菌量为10²～10⁴;0℃下分离计数,发现低温下微生物的数量较中温的多,每克湿土壤含菌为10³～10⁶。对该地区烃氧化菌进行了特别调查,以长城站为中心东西宽0．4Km,南北长1Km的范围内烃氧化菌发育最为活跃,但距长城站距离越远,烃氧化菌数量越少。     

溶解氧对单级自养脱氮系统功能菌数量的影响

摘要：【目的】研究溶解氧（Dissolved oxygen, DO）对单级自养脱氮系统功能菌数量的影响,为系统运行操控提出理论指导。【方法】从不同DO水平下的单级自养脱氮反应器中,分别提取活性污泥及生物膜样品基因组DNA,通过特异引物扩增系统内亚硝化菌（Ammonia oxidizing bacteria, AOB）、硝化菌（Nitrite oxidizing bacteria, NOB）及厌氧氨氧化菌（Anaerobic ammonia oxidizing bacteria, ANAMMOX）基因序列,PCR产物经回收克隆测序后,证实扩增产物为AOB、NOB及ANAMMOX 16S rDNA 保守序列,以含该序列的重组质粒作为定量PCR标准品。用荧光定量PCR技术对单级自养脱氮系统中各类功能菌进行定量分析。【结果】高DO有利于亚硝化菌AOB及硝化菌NOB生存,同时,活性污泥中AOB、NOB数量多于生物膜。DO对厌氧氨氧化菌ANAMMOX数量影响明显,高DO浓度将对ANAMMOX数量产生直接抑制,低DO浓度水平时,由于系统内缺乏厌氧氨氧化反应的电子受体NO3-或NO2-,也将间接影响ANAMMOX数量。【结论】本试验研究条件下,DO为(曝气)2.0/(停曝) 0.4 mg/L时系统运行效能最佳,ANAMMOX数量最多,AOB、NOB及ANAMMOX在此时构成一个协同代谢的稳定状态。     

三级串联垂直潜流人工湿地的脱氮性能及微生物学特征

探究了3种水力负荷(HLR)下三级串联垂直潜流人工湿地(T-VFCWs)对农村生活污水的处理效果,并解析了系统中的氮素转化机制.结果表明:当系统HLR由0.10增至0.20 m3·m-2·d-1时,T-VFCWs始终保持着对农村生活污水高效的处理效果,系统出水水质满足《城镇污水处理厂污染物排放标准》(GB 18918-...     

The CANON system (Completely Autotrophic Nitrogen-removal Over Nitrite) under ammonium limitation: interaction and competition between three groups of bacteria

The CANON system (Completely Autotrophic Nitrogen Removal Over Nitrite) can potentially remove ammonium from wastewater in a single, oxygen-limited treatment step. The usefulness of CANON as an industrial process will be determined by the ability of the system to recover from major disturbances in feed composition. The CANON process relies on the stable interaction between only two bacterial populations: Nitrosomonas-like aerobic and Planctomycete-like anaerobic ammonium oxidising bacteria. The effect of extended periods of ammonium limitation was investigated at the laboratory scale in two different reactor types (sequencing batch reactor and chemostat). The lower limit of effective and stable nitrogen removal to dinitrogen gas in the CANON system was 0.1 kg N m(-3) day(-1). At this loading rate, 92% of the total nitrogen was removed. After prolonged exposure (> 1 month) to influxes lower than this critical NH4+-influx, a third population of bacteria developed in the system and affected the CANON reaction stoichiometry, resulting in a temporary decrease in nitrogen removal from 92% to 57%. The third group of bacteria were identified by activity tests and qualititative FISH (Fluorescence In Situ Hybridisation) analysis to be nitrite-oxidising Nitrobacter and Nitrospira species. The changes caused by the NH4+-limitation were completely reversible, and the system re-established itself as soon as the ammonium limitation was removed. This study showed that CANON is a robust system for ammonium removal, enduring periods of up to one month of ammonium limitation without irreversible damage.     

Microbial Processes of Heavy Metal Removal from Carbon‐Deficient Effluents in Constructed Wetlands

This paper reviews the main microbial processes involved when toxic metals are removed from wastewater in constructed wetlands. Microbial activity is thought to play a key role in the detoxification of these metals. The paper concentrates on the microbial processes which affect the mobility, the toxicity and bioavailability of metals, namely biosorption, metal sulfide precipitation by sulfate reducers, redox transformations, and methylation, as well as microbe‐plant interactions. These reactions result in either the precipitation and accumulation of metals in wetland soils, or their volatilization and emission into the atmosphere. The possibilities of optimizing the microbially mediated reactions for the development of wetland technology are discussed as a long‐term metal retention strategy.     

Coupling of Microbial Processes of Methane and Ammonium Oxidation in Soils

The effect of ammonium ions on the activity of methane oxidation in soils was studied. The degree of inhibition of the methanotrophic activity in the presence of ammonium in the soil solution was quantitatively assessed as dependent on ammonium concentration and the properties of different types of soils of the European part of Russia.     

OLAND生物脱氮系统中硝化菌群16S rDNA的DGGE分析

为了考察生物脱氮系统中硝化菌群（氨氧化菌和亚硝酸氧化菌）的种群多样性及硝化菌群随溶解氧降低的种群变化规律,并建立一套行之有效的用于自养生物脱氮系统中功能微生物菌群的快速分子检测技术,采用DGGE（变性梯度凝胶电泳）分子检测技术对硝化菌群的16SrDNA的特异性PCR扩增产物进行了分析,结果表明：OLAND生物脱氮系统中氨氧化菌和亚硝酸氧化菌随溶解氧的降低表现出了不同的种群变化规律,氨氧化菌种群多样性受溶解氧的影响非常大,而非亚硝酸氧化菌的种群多样性比较单一,且不受溶解氧的影响。结合FISH（全细胞荧光原位杂交）分析结果表明,在OLAND限氧稳定运行后期,亚硝化单胞菌属（Nitrosomonas）是主要的氨氧化菌,占OLAND限氧亚硝化阶段反应器中总细菌数的72.5%左右。     

Change of Microbial Populations in a Suspended-sludge Reactor Performing Completely Autotrophic N-removal

Summary In recent years, several novel processes for N-removal almost without consumption of organic carbon under oxygen-limited conditions have been discovered, which may be a promising option for low-cost N-removal from ammonia-rich wastewater. In this study, a laboratory scale suspended-sludge reactor was continuously operated under low dissolved oxygen concentration. High N-removal efficiency and subsequently degradation of the reactor were observed. Molecular analysis based on a partial-16S rRNA gene library showed that, at the stage of high efficiency, the biomass was composed of Planctomycete-like bacteria (up to 40%) and heterotrophic organisms (approximately 60%) as well as a few ammonia-oxidizing bacteria and at the stage of degradation, the autotrophic ammonia-oxidizing bacteria were dominant (up to 70%) and Planctomycete-like bacteria were no longer found in the sludge. Three specific Planctomycete-16S rRNA-targeted probes were used for fluorescence in situ hybridization (FISH). The results showed that at the high-efficiency stage, Planctomycete-like bacteria, present at approximately 20% of the total bacteria, lay frequently in the middle of flocs, while the heterotrophic bacteria occurred within the outer layers. This work revealed that the change of the microbial populations is the key reason for reactor deterioration, and the heterotrophic bacteria probably play an important role in sustaining the biomass structure of the sludge.     

Ammonium versus Nitrate Nutrition of Plants Stimulates Microbial Activity in the Rhizosphere

Using an alkaline calcareous soil, pot experiments were conducted to elucidate the effects of NH ₄ ⁺ vs. NO ₃ ⁻ nutrition (50 or 100 mg kg⁻¹ soil) of wheat and maize on microbial activity in the rhizosphere and bulk soils. Dicyandiamide was used as nitrification inhibitor to maintain NH ₄ ⁺ as the predominant N source for plants grown in NH ₄ ⁺ -treated soil. While maize grew equally well on both N sources, root and shoot growth of wheat was higher under NH ₄ ⁺ than under NO ₃ ⁻ nutrition. Bacterial population density on roots, but not in the rhizosphere soil, was higher under NH ₄ ⁺ than under NO ₃ ^– supplied at 150 mg N kg⁻¹ soil; whereas at both N levels applied, NH ₄ ⁺ compared to NO ₃ ⁻ nutrition of wheat and maize significantly increased microbial biomass in the rhizosphere soil. Under both plant species, NH ₄ ⁺ vs. NO ₃ ⁻ nutrition also increased aerobic and anaerobic respiration, and dehydrogenase activity in the rhizosphere. As microbial activity in the planted bulk and unplanted soils was hardly affected by the N-source, we hypothesize that the stimulation by NH ₄ ⁺ of the rhizosphere microbial activity was probably due to higher availability of root exudates under NH ₄ ⁺ than under NO ₃ ⁻ nutrition.     

强化生物除磷系统的微生物种群及其表征技术

详细介绍了强化生物除磷系统(enhanced biological phosphate removal,简称EBPR)中的微生物种群及其表征技术,提出了研究EBPR中微生物种群及其表征技术的发展方向。     

Characterization of Bacterial Flora from Afşin–Elbistan Lignite Mine for Potential Microbial Biotech Applications

Abstract

The energy demand is enhancing day by day all over the world with the increasing of huge population. Lignite, a fossil fuel with low energy efficiency, is used in many different fields. However, lignite is primarily an energy source and its use without treatment causes environmental pollution as well as low energy efficiency. Advantages are known for the use of microorganisms in lignite processing technologies, and research is underway to improve this process. In this research designed to meet this need, isolation and characterization of bacteria with potential for use in biotechnological applications resistant to extreme conditions were performed from lignite mine. Turkey’s largest lignite mine bacterial flora have been identified for the first time in this study. In addition to lignite processing technologies, a bacterial strain collection has been created to be used in other biotechnological studies. This collection of bacteria includes Methylorubrum zatmanii, Streptomyces argenteolus, Streptomyces galilaeus, Pseudomonas mandelii, Paenarthrobacter aurescens, Paeniglutamicibacter sulfureus, Arthrobacter humicola, Bacillus pumilus, Bacillus subtilis, Bacillus safensis, Kocuria polaris, Arthrobacter sp., Rhodococcus sp. This specific bacteria collection is open to sharing with other researchers who want to do field applications.