159 resultados para Ammonia - Volatilization


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组特殊自养氨氧化混合种群,表现:无机环境种群生长迅速、生物量高;在一个完全无机的自养生长环境中,不仅保持高氨氧化速率,并出现丰富的异养微生物种群;该种群置于异养、厌氧环境中,迅速表现出产氢特征。对于这样一个特殊的生态体系,研究其共生机理,以及联接这些种群之间的碳源和能源问题,将具有非常重要意义。我们拟从种群特征、细胞表面分泌产物、游离体系产物多糖、蛋白和脂肪酸方面开展研究。 第一部分,自养氨氧化混合种群的基本特征。采用氨氧化培养基,进行种群氨氧化特征研究;采用扫描电镜观察自养混合种群的微观特征;沉降、离心去除微生物种群,分析水相中的总有机碳、糖类等物质;利用LB培养基进行种群的分离、纯化,并采用DGGE手段对微生物种群结构进行分析。结果表明,接入菌种后(2/5000(V/V)),培养液中氨(200mg/L)在3-5天内快速降解;亚硝酸盐与氨氮变化呈负相关趋势,仅有少量硝酸盐含量(< 30mg/L)。氨氧化种群的生物量增长与氨氧化趋势一致,初始生物量7.75 mg/L(蛋白含量),3-5天后生物量快速增长,并达到最高63.06 mg/L(蛋白含量)。电镜图片显示,种群外包裹一层粘液。离心除去菌体后,检测培养液总有机碳和糖的含量,同样表现出与生物量增长相似的特征,分别由初始的3.73、2.35 mg/L,3-5内天迅速增加,并分别达到最大值35.19、27.45 mg/L。经初步分离、纯化并对纯化菌株进行测序,获得了10株异养微生物分别为布鲁氏菌科苍白杆菌属、纤维单孢菌、类芽孢菌属、黄杆菌属、无色杆菌、鞘脂单胞菌、嗜麦芽寡养单胞菌、噬氢菌属、硫红球菌、假单胞菌;DGGE显示,约有20分条离带,我们对其中的两条优势条带进行切割回收测序,鉴定为欧洲亚硝化单胞菌(Nitrosomonas eur)。 第二部分:混合种群自养-异养菌共生的可能机制。在对微生物种群特征初步分析基础上,针对胞外糖类组分可能被微生物代谢分解,我们重点对微生物细胞蛋白质与糖类进行分析。采用超声结合RIPA裂解液裂解,SDS-PAGE电泳分析混合种群总蛋白种类,并通过氨基酸分析仪及红外光谱法分析氨基酸组成及蛋白红外特征。采用超声破碎结合反复冻融对细胞样品进行处理,提取液采用醇沉、Sevage脱氮白,凝胶过滤方法脱盐和分级分离。对提取物的糖分析包括:紫外扫描,红外光谱,核磁共振,单糖组成分析;扫描电镜观察菌群破裂现象。SDS-PAGE分析结果表明:氨氧化种群不同生长阶段都显示出42kD蛋白表达量很高,d4时42kD蛋白表达已经很强,4-7d内一直持续这种过量表达,直到d8后表达开始减弱。说明42kD蛋白可能与氨氧化密切相关。红外光谱分析显示:细胞提取物的特征峰分布在3427.42cm-1、1718.18 cm-1和1681.72 cm-1、1160.07和1086.74 cm-1,分别对应为OH、 C=O、C-O-C基团,表明具有蛋白的典型特征;氨基酸分析显示蛋白中的Gly,Asp,Ala,Glu含量相对较高。 提取物中胞外多糖分离谱图得到不均一组分,共得到6个收集峰;紫外扫描在201-213 nm处有多糖吸收峰,同样表明多糖成分不均一性;多糖红外光谱特征峰主要分别在3400.49 cm-1、2920.28 cm-1、1154.54和1087.52 cm-1,对应OH、-CH2- or CH 、C-O-H or C-O-C等多糖特征基团;多糖提取物核磁共振1H d4.3~5.9之间出现强吸收峰,这是1H中,多糖存在的明显证据,1H NMR中,其中O-乙酰基的甲基上的氢信号为d1.1~1.3之间。糖肟全苯甲酸酯衍生物的HPLC测定中,得到单一的单糖峰,由于时间问题,还未进行更深入的试验;电镜图片显示,种群中的细胞有大量的破裂现象。 实验表明,自养氨氧化混合种群显示出快速的氨氧化速率,氨氧化过程生物量和有机质的增加明显。微生物种群包裹粘液层,并分离纯化出大量的异养菌;去除菌体后的游离培养液中存在有机质(包括多糖)说明无机自养生长体系中存在异养菌生长、繁殖的二次碳源;细胞提取物中蛋白条带数目多、种类丰富;细胞多糖提取物具有明显的多糖特征,以及单糖的存在。结合种群的显微特征和游离体系中的有机质的检测结果,我们认为,无机自养生长体系中,种群细胞生长过程中发生的破裂现象可能是导致大量的蛋白、多糖释放到游离胞外,并成为其他异养菌生长的碳源和氮源。这可能是自养体系中,大量异养菌共生的可能机制,至于是什么原因引起种群生长过程中产生的破裂现象,还有待下一步深入研究。 A group of mixed autotrophic ammonia oxidizing populations, having much biological characteristic tested by concerned personnel for pilot test: Performed rapid population growth and obtained high biomass in inorganic environment; Not only maintained a high rate of ammoxidation, promoted a wealth of heterotrophic microbial populations growth in a totally inorganic and autotrophic growth environment; Placed in heterotrophic and anaerobic environment,had the performance characteristics that could rapidly produce hydrogen.For such a special ecological system, Study its symbiotic mechanism and the connection between these populations of carbon and energy issues, will have a very important significance. We intended from the characteristics of the population, the secretion product of cell surface, free substance in the liquid medium like polysaccharide, protein and fatty acids carrying out research. Part I: The basic features of mixed autotrophic ammonia oxidizing populations . Use inorganic liquid medium, processed study for ammonia oxidation characteristics of the population; we used scanning electron microscopy to get micro-features of autotrophic ammonia oxidizing populations .The medium was carried out settlement and centrifugal then removed the microbial populations, after all of that we analysis the water phase for total organic carbon(TOC), carbohydrate and other substances; Solid ammonia oxidizing medium was adopted to separation and purification of population, DGGE means was for structure analysis of microbial population. The results showed that after the inoculum of bacteria (2 / 5000 (V / V)), ammonia in the culture medium (200 mg / L) was rapid degradation in 3-5 days; ammonia and nitrite have the negative correlation between changes in the trend, then only a small amount of nitrate content (<30mg / L). The biomass growth of ammoxidation population in line with the trend of ammonia oxidation, the initial volume of it was 7.75 mg / L (protein content), in 3-5 days upto 63.06 mg / L (protein content). Electron microscope image showed, the populations were wrapped in a layer of mucus, including the a large number ruptted micorbe , Centrifuge to remove bacteria, then detected the medium for total organic carbon and sugar content, result took on the same characteristics with biomass growth, that were from the initial 3.73、2.35 mg / L respectively, in 3-6 days achieved rapid increase in the maximum to 35.19、27.45 mg / L respectively. After initial separation、 purification ,then processed sequencing to strains purified and got the result that there were 10 heterotrophic microorganisms : Brucella Branch pale bacillus, Cellu lomonas, Bacillus species category, a Flavobacterium, colorless Bacteria, Aeromonas sheath fat, little support maltophilia Aeromonas, macrophages species hydrogen, sulphur-MI, Pseudomonas bacteria spores; DGGE display, there were 20 separation bands approximately. Part II: Mixed populations that autotrophic - heterotrophic bacteria symbiotic mechanism. On the basis of preliminary analysis of microbial population characteristics, aiming at extracellular carbohydrate components might be decomposition by microbial, we focused on microbial cell protein and carbohydrate analysis. Using ultrasound combined with RIPA lysis cracking the cells, SDS-PAGE electrophoresis analysis the total protein species of the population, and through the amino acid analyzer studied the compositions of amino acid and infrared spectroscopy analysis of a protein infrared characteristics. Using ultrasound combined with repeatedly freezing and thawing to treated the cell sample, then took the means that alcohol precipitation, deproteinization by Sevage, gel filtration aimed at desalination and grade separation to deal with the lysates . The extraction of sugar analysis included: UV scanning, IR, NMR, single-sugar composition analysis. SDS-PAGE analysis showed that: 42 kD protein expression was very high at different growth stages of mixed autotrophic ammonia oxidizing populations , on the fourth day, 42 kD protein expression had been very strong, 4-7d, it had continued this excessive expression, then started to weaken after 7 days. 42 kD protein that might be closely associated with ammonia oxidation. Infrared spectral analysis showed that: cell extracts with the characteristic that the peak distribution in 3427.42 cm-1、1718.18 cm-1 and 1681.72 cm-1、1160.07 cm-1 and 1086.74 cm-1 corresponding to OH、C = O、C-O-C Groups which had the typical characteristics of protein; and analysis showed that amino acids including Gly, Asp, Ala, Glu ,the content in the protein is relatively high. Exopolysaccharide in the extracts had the separation map that it was uneven, received a total of six collection peaks by the detection mode of phenol-sulphruic acid method ; ultraviolet scan in the 201-213 nm department had polysaccharide absorbing peak, the same ingredients that polysaccharide heterogeneity; infrared polysaccharide spectral characteristics of the main peak at 3400.49 cm-1, 2920.28 cm-1, 1154.54 and 1087.52 cm-1, corresponding OH,-CH2-or CH, C-O-H or C-O-C;and other characteristics of polysaccharide group; 1H NMR of polysaccharide extract appeared absorption peak between d4.3 ~5.9, which is the apparent evidence of polysaccharide, In 1H NMR, the hydrogen signal of one of O-acetyl was between 1.1 to 1.3. The determination of Sugar oxime whole benzoate derivatives by HPLC, there was a single-sugar peak, as a matter of time, yet more in-depth test. Summary: Mixed autotrophic ammonia oxidizing populations show us that it had the ability in ammonia oxidizing and it was great, organic matter and biomass increased significantly in the process of ammonia oxidation. Microbial populations was wrapped up slime layer, the phenomenon of cell breakdown obviously, and there were a lot of separation and purification of the heterotrophic bacteria; a lot of organic matter (including polysaccharides)remined in the medium that removal of cell indicated the inorganic system existed secondary carbon sources that could be used by the heterotrophic bacteria ; there were a large number proteins bands of cell extract, rich variety; cell extracts of polysaccharide had obvious characteristics of polysaccharide, and the existence evidence of single-sugar. Combined population of microscopic characteristics and free of organic matter in the test results, we believe that the health of inorganic system, population growth occurred in the course of the breakdown of the phenomenon is likely to lead to a lot of protein and polysaccharide released into the extracellular free, And other heterotrophic bacteria use them to the growth as carbon and nitrogen. This may be autotrophic system, the large number of heterotrophic bacteria symbiotic mechanism.

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猪场废水COD浓度高、氨氮浓度高、悬浮物浓度高,已成为农村面源污染的主要来源,并严重威胁到农村饮用水安全。猪场废水氨氮浓度高、处理难度大,如何采用经济高效的方法,去除氨氮使其达到排放标准,一直是猪场废水处理中面临的重要难题。 厌氧氨氧化是近年受到国内外水处理研究者广泛关注的新型生物脱氮技术,具有不需要外加有机碳源、节省供氧量、降低能耗等优点。虽然国内外研究者对厌氧氨氧化过程的脱氮机理、厌氧氨氧化菌的生理生化特性等进行了多方面的研究,但已有的报道大多以模拟废水为研究对象,以猪场废水为研究对象的报道,在国内外文献中极少有报导。 本论文以猪场废水为主要研究对象,考察了猪场废水的亚硝化过程、厌氧氨氧化的启动过程,并对亚硝化和厌氧氨氧化联合用于猪场废水脱氮进行了探索。 1.论文首先研究了猪场废水的亚硝化过程,考察了废水水质和主要运行条件对亚硝化过程的影响。实验表明:(1)亚硝化阶段反应时间为8到10h时,出水中氨氮和亚硝酸盐浓度比可达到1:1~1:1.23,满足厌氧氨氧化反应对二者比例的要求;达到前述要求时,氨氮去除率达到58.3~65.6 %,亚硝化率在整个过程均保持在97 %以上,COD去除率在59.2~68.6 %;(2)曝气量(溶解氧)对亚硝化过程影响显著,随着曝气量增大,达到厌氧氨氧化要求的氨氮与亚硝酸盐氮浓度比例所需水力停留时间τ越短,pH出现明显下降的时间越短;(3)τ对应的pH在7.8~8.1之间,无需进行pH调节即可满足厌氧氨氧化反应对pH的要求;(4)氨氮和COD降解过程遵循一级反应动力学,氨氮和COD降解的速率常数分别为0.0656~0.0724 1/h和0.0491~0.0664 1/h。 2.在进行亚硝化过程研究的同时,以模拟废水为试验对象,进行厌氧氨氧化启动研究。以反硝化污泥和养殖厂储水池厌氧底泥的混合污泥作为接种污泥,历时大约100天,培育出具有厌氧氨氧化活性的污泥,氨氮和亚硝酸盐氮最高进水浓度分别为223.8 mg/L和171.4 mg/L,去除率最高分别达48%和41.5%,此时二者消耗比例为1.33:1。 3.在猪场废水的亚硝化研究完成和厌氧氨氧化过程初步启动成功后,在模拟废水中逐步加入猪场废水的亚硝化处理出水,逐步实现亚硝化和厌氧氨氧化的组合。亚硝化出水添加到厌氧反应器后,厌氧氨氧化反应仍可继续进行,且去除效率逐步提高。研究发现添加的亚硝化出水中携带的亚硝化细菌在厌氧氨氧化菌膜外层生长并累积,增加了厌氧氨氧化反应基质的传质阻力,妨碍了厌氧氨氧化效率的提高。 4.亚硝化-厌氧氨氧化实际工程应用探索中,生物接触氧化池可在有效去除废水中的有机物的同时实现亚硝化,出水中氨氮和亚硝酸盐比例平均为1.10,可满足后续厌氧氨氧化的要求;在适宜的进水浓度和温度下,ABR池出现了厌氧氨氧化启动的迹象;研究同时发现,水质的波动和气温的变化是工程中影响厌氧氨氧化菌活性的重要因素。 论文的主要创新点在于:(1)以猪场废水为研究对象,以实现厌氧氨氧化为目标,对亚硝化过程进行了比较详细的考察,获得了亚硝化出水满足厌氧氨氧化要求的工艺条件,通过对其COD和氨氮降解过程的考察,得出亚硝化阶段COD降解和氨氮去除的动力学模型;(2)对亚硝化-厌氧氨氧化处理猪场废水进行了探索,确立了影响其污染物去除率稳定的重要因素。 论文的上述研究成果,为厌氧氨氧化技术的实用性研究提供理论依据。 Piggery wastewater, which is characterized by high concentration of COD、ammonium and suspend substance, has become a most important source of non-point source pollution and also severely threats drinking water security in rural area. How to discharge piggery wastewater with the ammonium concentration meeting standard by economical and effective method? This is the most urgent problem in piggery wastewater treatment. As a new biological nitrogen removal technology, Anammox process has been paid more and more attention by researchers all over the world. Anammox has advantages of no need of organic carbon addition, low oxygen consumption and energy consumption. Plenty of investigations have been carried out to the mechanism, physiological and biochemical characteristic of bacteria about Anammox. Most of researches focused on synthetic wastewater, there is rare report about its application in piggery wastewater. In this paper,experimental studies were performed to investigate Sharon process in treatment of piggery wastewater,the start up process of Annammox using synthetic wastewater were studied, the feasibility of applying Sharon-Anammox process in the nitrogen removal of piggery wastewater was evaluated. 1. Sharon process of piggery wastewater was firstly investigated to analyze the effects of water quality and main running parameters, which meet the NH4+-N to NO2--N ratio requirement of successive Anammox. Results showed: (1)During Sharon Process,after 8~10 hours’ reaction the NH4+-N to NO2--N ratio in effluent reached 1:1.0~1:1.23, when the removal percentage of NH4+-N was 58.3~65.6 %, a semi-nitration rate of above 97 % was achieved during the process; meanwhile 59.2~68.6 % of the COD was also removed. (2)The aeration rate(oxygen) had obvious effect on the hydraulic retention time(τ) which met the NH4+-N to NO2--N ratio requirement of Anammox. As aeration rate increased, the hydraulic retention time(τ) was shortened. (3) The pH corresponding to τ was between 7.8 and 8.1, thus it needed no artificial adjustment. (4) The reduction of ammonia and COD followed the first-order reaction kinetics. The velocity constants of ammonia and COD were 0.0656~0.0724 1/h and 0.0491~0.0664 1/h, respectively. 2. The startup of Anammox process using the artificial wastewater was performed simultaneously with Sharon. The aim was to investigate the running parameters of Anammox and make foundation for the combination stage. By using the mixture of denitrifying sludge and anaerobic sludge in tank of the breeding factory, sludge of Anammox activity was cultivated in UASB after 100 days. The removal percentage of NH4+-N and NO2-N were up to 48% and 41.5%, respectively, when the NH4+-N and NO2-N influent concentration were 223.8 mg/L and 171.4 mg/L, respectively, the NH4+-N and NO2-N removal rate was 1.33:1. 3. After investigation of Sharon and startup of Anammox, effluent of Sharon process was added into the synthetic wastewater to combine Sharon and Anammox step by step. It took some time after the addition of Sharon effluent that Anammox reaction continued and the removal rate kept increasing. It indicated that nitrifying bacteria were carried by the Sharon effluent cumulated in the outer layer of Anammox. This enhanced transfer resistance of Anammox reaction and the increasing removal rate was restrained. 4. In the bio-contact oxidation pond of practical project, Sharon process were carried out successfully and organic compounds were removed effectively. An average NO2-N/ NH4+-N rate of 1:1.0 was achieved in the effluent, which met the requirement of successive Anammox. Under condition of suitable influent concentration and temperature, there was evidence that Anammox could start up in ABR. The variety of wastewater and temperature had great affects on Anammox activity in practical engineering. Innovation of this paper: (1) The Sharon process for treating piggery wastewater was discussed in details. Technological parameters that met requirement of Anammox were obtained. The dynamic models of COD and ammonium removal in the process were educed. (2) Sharon-Ananmmox for treatment of piggery wastewater was investigated, and the primary influencing factors was studied. This paper could be a theoretical consult for research of Anammox utility.

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一体化反应器由于投资少、占地小、管理运行方便等优点而备受青睐。但现有的一体化反应器大都适用于处理中低浓度废水,耐受负荷普遍偏低。本课题研制出新型高效的厌氧好氧一体化生物反应器,旨在通过反应器结构优化、高效微生物载体研制、配合高效微生物菌剂技术处理中高浓度有机废水,实现高效和低耗,降低设备造价,提高反应器运行稳定性。 首先开展了菌剂对废水的适配试验。采用15种不同的微生物菌剂,以葡萄糖配水、中药提取废水、啤酒废水、氨氮配水为基质,分别测定了微生物菌剂的耗氧速率和厌氧比产甲烷速率,以其为指标比较了各菌剂对废水的适配性。根据结果选择活性高的14#、8#、10#菌剂,在试验室进行了菌剂对废水的连续处理试验,取得良好的处理效果,为菌剂在厌氧好氧一体化生物反应器的小试、中试中的应用奠定了基础。 经小试研究后,又对厌氧好氧一体化生物反应器进行了处理发酵废水的中试研究。试验结果表明,反应器启动快,系统有机负荷2.72 kgCODm-3d-1时整个反应器去除率保持在84.5%~93.19%,在30多天内一次启动成功。冲击负荷试验中,系统总有机负荷最高可达到8.88 kgCODm-3d-1,系统去除率稳定在88.10%~96.88%,说明反应器处理效率高,抗冲击能力强。稳定运行期间,COD去除率可达90%以上,各项指标都能达到国家排放标准。 此外,对反应器配套系统高效菌剂、高分子复合颗粒载体进行了研究。结果显示,菌剂与反应器适配良好,各功能区形成了丰富、高活性的微生物,厌氧区颗粒污泥TS高达83.9 gL-1,VS/TS为56.9%~57.4%,比产甲烷活性为280~350 mLCH4 gvss-1d-1;好氧区固定化微生物TS高达1.921 gL-1,VS/TS为94.02~94.30%。对载体性能的研究表明,此高分子复合颗粒载体密度适中,易于流化,不易流失;粗糙多空,易于挂膜;且无生物毒害作用,稳定安全,是一种优良的生物载体。反应器各功能区对废水的降解过程分析,说明了反应器、菌剂、载体适配良好,在其协同作用下,实现了污染物的高效降解。 The integrated reactors were popular because of their characteristics such as little investment, small occupation of land, convenient of manage and running etc. But the present integrated reactors were mostly applied for treating wastewater of low concentration, the load tolerance was generally on the low side. A new type integrated anaerobic-aerobic bio-reactor was developed, which was conducted to treating organic wastewater of middle or high concentration by optimization of reactor structure, development of efficient microbe carrier and adaptation of high active microbial blends, to achieve high efficiency and low consume, reduce equipment cost, enhance running stabilization of reactor. The adaptability test of microbial blends on different wastewater was carried on firstly. Oxygen consumption rate and anaerobic specific activity of methane producing of 15 different microbial blends were measured separately taking glucose artificial wastewater, Chinese herb extracting wastewater, brewery wastewater and ammonia nitrogen artificial wastewater as substrate, by which the adaptabilities of different microbial blends to wastewater were compared. According to the results high active microbial blends 14#, 8# and 10# were selected and used in the continuous treatment of wastewater in the laboratory and had obtained good effect, which had laid a foundation for application microbial blends to small scale test and pilot test of integrated anaerobic-aerobic bio-reactor. After the small scale test, the pilot test of the integrated anaerobic-aerobic bio-reactor treating fermentation wastewater was carried on. The test results showed fast initiation of the reactor. When system organic load reached 2.72 kgCODm-3d-1the COD removal rate of the reactor was stable between 84.5%~93.19% and it initiated successfully in more than 30 days at a time. In the load shock test the maximum organic load of system could reach to 8.88 kgCODm-3d-1 and the COD removal rate could be stable between 88.10%~96.88% which indicated that the reactor was efficient for treating wastewater and had strong resistance to shock load. At stable running period the COD removal rate of the reactor was over 90% and each index of wastewater could reach to the national discharge standards. In addition, the high active microbial blends and the macromolecule compound granule carrier, the matching system of the reactor was studied. It showed that the microbial blends adapted well to the reactor and abundant and high active microbes were formed in each functional field. The TS of granule sludge in anaerobic field was as high as 83.9 gL-1, the VS/TS was 56.9%~57.4%, the specific activity of methane producing was 280~350 mLCH4 gvss-1d-1. And the TS of immobilized biological granule was as high as 1.921 gL-1, the VS/TS was 94.02%~94.30%. Study on the carrier showed that the self-made macromolecular compound granule carrier was moderate of density, easy of fluidization, unease of running off, rough and porous, easy of films fixation, no bio-toxic, stable and safe, was a kind of superior carrier. Analysis of degradation process in each functional field confirmed the reactor, microbial blends and carriers were in good adaptation and wastewater was decontaminated by their cooperation.

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In this paper, bioconversion of trans-cinnamic acid(t-Ca)to L-phenylalanine (L-phe) has been investigated by using immobilized yeast cells with induced L-phe Ammonia-lyase(PAL, EC.4.3.1.5) as biocatalysts. The contents are the following. (1) Thirty strains of yeasts, including two genera (Rhodotorula, Sporobolomyces), six species (R. glutinis R. minuta,R.rubra,R.sineses,R.roseus and S.salmonicolor)were screened for their ability to converse the substrates, t-Ca and ammonia, to the product, L-phe, by using yeast cells as biocatalyst, and primary evaluation for PAL activity of the selected strains was investigated. From the results of the screening experiments, it was found that 22 strains were able to produce L-phe from t-Ca with the range of conversion yield from 2% to 67%. Studies on PAL formation time course during cultivation show that the maximum PAL activity of several different strains ranges from 2.3 to 14.4×10-3U/mg cell dry weight. The biomass of tested strains at their maximum enzyme activity is also greatly varied. (2)One of the selected strains, R. rubra as 2.166, was used for immobilized cells as biocatalysts to produce L-phe. The optimum conversion conditions and effective stablization agents were investigated. The results shown that polyacrylamide gel was chosen as a suitable matrix for immobilization of the yeast cells, and it can retain 88% of the PAL activity in the reverse direction at the following reactive conditions: [t-Ca]: 34mM. [NH4OH]: 6.OM.PH10.00, temperature: 30℃. (3) The effects of various kinds of effectors on the production of L-phe were also examined. Membrane permeabilizing agents can stimulate L-phe synthesis, but make the stability of PAL decline greatly. Polyalchoholic agents and glutamic acid were very effective for the stabilization of PAL. At the presence of glutamic acid (5%), the half life of L-phe productivity with the immobilized cells was extended to 192 hours, which was much higher than most of that having been reproted, while the half life of resting cells was only about 15 hours. (4) Use of initial velocity studies on the kinetics of enzyme-catalized reaction indicated that the apparent Km value was 13.0mM for the immobilized cells, and 4.8mM for the resting cells. Thermostability of the immobilized cells was better than the resting cells. Fluid bed bioreactor is more effective than batch bioreator in prolonging the thermostability of the biocatalysts. (5) CGA- 688 resin column chromatographic procedure was employed in the isolation and purification of L-phe, t-Ca and other substances from the reactire mixture. (6) Preparative-scale production of L-phe on a level of gram amount by immobilized cells from the culture broth of R. rubra AS2.166 allowed for the conversion yield with 30%. The characteristic physico-chemical criteria (including melting point, optical activity, elements analysis, IR, NMR) are the same with the standard L-phe. 本文报告了利用诱导的苯丙氨酸解氨酶 (PAL.EC.4.3.1.5)催化反式肉桂酸(t-Ca)氨加 成制备L-苯丙氨酸(L-phe)的研究,主要内容为:(1) 我们搜集了三十株酵母菌株,利用全细胞转化t-Ca生成L-phe的能力进行了直 接筛选,并对其PAL活性水平进行了初步评估研究。研究结果表明,其中22株酵母具有转化t-Ca生产L-phe的能力,它们包括 Rhodotorula glutinis,R.rubra, R.sineses 和Sporobolomyces roseus 的菌株,转化率在2-67%。细胞生长和PAL形成过程的研究 表明,不同菌株PAL最大活力在2.3-14.4×10-3U/mg 细胞干重,达到最大PAL活性时各株酵母的生长情况也极不一致。(2) 利用筛 选出的一株深红酵母R.rubra AS2.166 作为供试菌株,研究了细胞固定化条件下生物转化的最适条件及PAL在固定化条件下的稳定 性。结果表明以聚丙烯酰胺凝胶包埋法较为理想,能使细胞合成L-phe活力保持88%,最适t-Ca浓度为34mM,最适NH4OH浓度为6M,最 适PH10.0,最适温度45℃。(3) 多种效应物对L-phe 合成的影响研究表明:表面活性剂能刺激L-phe的合成,但使PAL稳定性下降。 多羟基化合物及Glu对PAL的稳定十分有效在有Glu存在下,能使固定化细胞合成L-phe的半寿期达192小时左右,高于大部分现已报 导的固定化结果。(4) 用初速度法研究了深红酵母AS2.166中PAL的酶促反应特征,测得固定化细胞对t-Ca的表观米氏常数Km为 13.0mM,全细胞为4.8mM,细胞固定后热稳定性提高。(5) 建立了适合低浓度分离纯化产物与底物的聚苯乙烯大孔树脂柱层析技术 ,能使L-phe与t-Ca及产物混合物中其它成分有效分开。(6) 利用固定化的R.rubra AS2.166细胞所做的制备实验能够使L-phe的产 率达到30%左右,其主要的理化指标(包括熔点、比旋光度、元素分析、IR、NMR等)与标准L-phe一致。

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垃圾卫生填埋是国内外城市垃圾的主要处置方法。垃圾渗滤液是渗入填埋场垃圾的降水混合垃圾降解过程中产生的物质而形成的混合物,是垃圾填埋场向环境排放的主要污染物。渗滤液由于其所含高浓度有机和无机污染物,且其中很多物质有生物毒性或难生物降解,难于治理。特别是到填埋晚期,渗滤液中高浓度的氨氮更是增加了治理的难度。渗滤液场外硝化-原位反硝化是填埋场氮管理的新途径。本文利用从环境中筛选出优势硝化功能菌对渗滤液中的高浓度氨氮进行生物硝化,经硝化后的渗滤液回灌至以垃圾柱模拟的生物反应器填埋场,在填埋场内实现原位反硝化。 上述目标通过以下两部分来实现: 第一部分:渗滤液场外硝化。首先从污水厂的硝化污泥中富集并筛选出硝化功能菌,在模拟氨氮废水中优化。将驯化的硝化功能菌接种于连续式完全混合反应器(CSTR)进行高氨氮渗滤液硝化研究。在200余天的连续运行中,反应器硝化和有机物去除效果良好。在最大氨氮负荷和有机物负荷分别为0.65 g N l-1 d-1 和3.84 g COD l-1 d-1时,氨氮和COD去除率分别高于99%和57%。实验过程中发现,游离氨(FA)和溶解氧(DO)浓度对反应器中亚硝酸盐的积累影响很大。 第二部分:渗滤液原位反硝化。本文利用一个垃圾填充柱模拟生物反应器填埋场,研究了硝化渗滤液回灌对垃圾降解的影响,和回灌的硝化渗滤液中TON(总氧化态氮)对填埋场生物反应器产甲烷作用的影响。最后利用变性梯度凝胶电泳(DGGE)分析了硝化渗滤液回灌对垃圾填埋场菌群结构的影响。结果表明:回灌的TON被完全还原,反硝化为主要反应,最大TON负荷为28.6 mg N kg-1 TS d-1。当垃圾柱TON负荷大于11.4 mg N kg-1 TS d-1时,出现了产甲烷抑制,抑制作用随TON负荷的增加而加强。在此过程中,反硝化逐渐代替产甲烷作用成为填埋场内垃圾降解的主要反应,且更多产生的是清洁的氮气,而非温室气体甲烷。直到实验结束时,回灌硝化渗滤液的垃圾柱的甲烷产量仅相当于对照的2.5%,并且回灌的硝化渗滤液还加速了填埋场垃圾的降解与稳定。通过DGGE进行菌群结构分析发现,由于TON对填埋场的长期作用,反硝化菌增多而产甲烷菌减少。 Landfill still remains the chief method for MSW management around the world. Leachate is a mixture of rainfall permeating through landfill and organic and inorganic matters generated during decomposition of the wastes in the landfills, characterized as highly complicated and refractory wastewater. Ex-situ nitrification and sequential in-situ denitrification represents a novel approach to nitrogen management at landfills. In the present paper, nitrification was carried out in a continuous stirred tank reactor (CSTR) inoculated with nitrifying bacteria which were isolated from municipal WWTP of Chengdu city. The nitrified leachate from CSTR was recirculated to a lab-scale municipal solid waste (MSW) column where in-situ denitrification took place. The above object was achived through two parts as following: First, ex-situ nitification of leachate. After acclimated in simulated wastewater for 3 month, nitrifying bacteria isolated from WWTP nitrifying sludge were added to the CSTR for nitrification. The results over 200 days showed that the maximum nitrogen loading rate (NLR) and the maximum organic loading rate (OLR) was 0.65 g N l-1 d-1 and 3.84 g COD l-1 d-1, respectively. The ammonia and COD removal was over 99% and 57%, respectively. Moreover, the effects of free ammonia (FA) and dissolved oxygen (DO) on nitrification were investigated. Second, in-situ denitrification was studied in a municipal solid waste (MSW) column. Variation of nitrified leachate and its effects on the decomposition of municipal solid waste (MSW) were studied in a lab-scale MSW column to which nitrified leachate was recirculated. Additionally, DGGE was employed to investigate the microbial community of both MSW columns. The results suggested: complete reduction of total oxidized nitrogen (TON) was obtained with maximum TON load of 28.6 mg N kg-1 TS d-1 and denitrification was the main reaction responsible. Methanogenesis inhibition was observed while TON load was over 11.4 mg N kg-1 TS d-1 and the inhibition was enhanced with the increase of TON load. Denitrification gradually took over methanogenesis to become the main reaction responsible for decomposition of MSW while nitrogen gas, a clean byproduct, was generated instead. Till the end of the experiment, the average weekly methane production in the denitrification column was as low as 2.5% of that of the control, and the rate of decompition and stability of MSW was accelerated by the recirculation of the nitrified leachate.Owing to long term exposure of nitrified leachate to landfill, denitrifying bacteria increased and methanogen decreased.

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自养硝化过程在自然界氮素循环和污水处理系统脱氮过程中起着关键作用。因此,了解有机碳对硝化的影响和硝化菌与异养菌之间的竞争对微生物生态学和污水处理系统设计都很重要。目前对氨氧化到硝酸盐氮过程的研究文献很多,但对亚硝酸盐氧化过程在异养菌的存在下如何受到有机碳影响的研究甚少。本文从生理生化指标、基因组学、蛋白组学三方面考察了在实验室条件下有机碳(乙酸钠)对硝化细菌和异养菌组成的混合菌群的硝化性能、菌群结构及代谢功能的变化的影响。 全文分为两大部分: 第一部分为乙酸钠对游离态硝化混合菌群的硝化性能和菌群结构的短期影响。混合菌株先在自养条件下进行连续培养,两个月后硝化速率达到20 mg N/(L·d);而后离心收集菌体进行批式实验。在批式反应器中,初始亚硝氮均为126mg N/ L,乙酸钠-C 与亚硝酸盐-N 的比分别为0,0.44,0.88,4.41,8.82。结果表明:在低C/N 比(0.44 和0.88)时,亚硝酸盐去除速率比C/N=0 下高,细菌呈现一次生长;而在高C/N 比(4.41 和8.82)时,出现连续的硝化反硝化,亚硝酸盐去除率仍比对照下高,细菌呈现二次生长。不同C/N 比下微生物群落明显不同,优势菌群从自养和寡营养细菌体系(包括亚硝酸盐氧化菌,拟杆菌门,α-变形菌纲,浮霉菌门和绿色非硫细菌下的一些菌株)过渡到异养和反硝化菌体系 (γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas stutzeri 和P. nitroreducens 占主导)。 第二部分为乙酸钠对硝化混合菌群生物膜的硝化性能和菌群结构的长期影响。接种富集的硝化混合菌群于装有组合式填料的三角瓶中,于摇床中自养培养;两个月后填料上形成生物膜的硝化速率达到20 mg N/ (L·d);而后进行长期实验,每12 小时更换混合营养培养基(亚硝氮约200 mg N/ L,C/N 比同上)。结果显示:相较于C/N 比=0 时的亚硝酸盐氧化反应来说,低C/N 比出现了部分的反硝化,而高C/N 比则是几乎完全的反硝化。与对照比,C/N=0.44 时亚硝酸盐氧化速率并未受乙酸钠的影响,反而上升了,但C/N=0.88 时亚硝酸盐氧化速率有所下降。菌群结构分析表明自养对照与混合营养下微生物群落的不同;PCR-DGGE未检测出混合营养下硝化杆菌的存在,而显示异养菌尤其是反硝化菌的大量存 在。荧光定量PCR 结果表明随C/N 比上升,硝化杆菌数量从2.42 × 104 下降到1.34× 103 16S rRNA gene copies/ ng DNA,反硝化菌由0 增加至2.51 × 104 nosZgene copies/ ng DNA。SDS-PAGE 的结果表明不同C/N 比下的蛋白组较为复杂且呈现一定的差异性。 有机碳对亚硝氮氧化及微生物群落的影响很复杂,本文分别讨论了对游离态和生物膜固定态两种状态的混合菌群相应的短期和长期影响研究。研究发现,有机碳并非一定带来硝化的负影响,如果控制在适当的C/N 比范围,有机碳是有利于亚硝氮氧化的。这些发现阐明了有机碳和硝化反硝化的关系,填补了硝化微生物生态学上的空白,对污水处理系统中减少异养菌的影响并提高氮去除率有一定理论指导意义。 Nitrification plays a key role in the biological removal of nitrogen in both nature and wastewater treatment plant (WWTP). So, understanding of the effect of organic carbon on nitrification and the competition between nitrifying bacteria and heterotrophic bacteria is important for both microbial ecology and WWTP design and operation. Despite the fact that the nitrification process of ammonia to nitrate has been extensively investigated, it is not known how the process of nitrite oxidization is affected by organic carbon when heterotrophic bacteria are present. By measuring different physiological and biochemical parameters, as well as using genomic DNA and proteome analysis, we investigated the influence of organic (acetate) on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria under laboratory conditions. The dissertation involves two parts: Part one deals with the effect of organic matter on functional performance and bacterial community shift of nitrite-oxidizing and heterotrophic bacteria under suspended state. The bacteria were prepared in a continuous-flow stirred reactor under autotrophic condition; after two months, the nitrification rate of the culture reached about 20 mg N/ (L·d); then the bacteria were harvested for the next batch experiments. The initial concentrations of nitrite were 126 ± 6 mg N/ L in all flasks, and sodium acetate (C) to nitrite (N) ratios were 0, 0.44, 0.88, 4.41, and 8.82, respectively. The results showed that at low C/N ratios (0.44 or 0.88), the nitrite removal rate was higher than that obtained under autotrophic condition and the bacteria had single growth phase, while at high C/N ratios (4.41 or 8.82), continuous aerobic nitrification and denitrification occurred besides higher nitrite removal rates, and the bacteria had double growth phases. The community structure of total bacteria strikingly varied with the different C/N ratios; the dominant populations shifted from autotrophic and oligotrophic bacteria (NOB, and some strains of Bacteroidetes, Alphaproteobacteria, Actinobacteria, and green nonsulfur bacteria) to heterotrophic and denitrifying bacteria (strains of Gammaproteobacteria, especially Pseudomonas stutzeri and P. nitroreducens). Part two describes the influence of acetate on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria in biofilms. Bacterial enrichments was transferred into flasks with polypropylene carriers and cultured under agitated and autotrophic condition. After two month, the biofilms grown on the carriers had a nitrification rate of about 20 mg N/ (L·h); then the biofilms were refreshed with mixotrophic medium (nitrite were 200 mg N/ L in all flasks, and C/N ratios was the same as above) every 12 h. the results show: normal nitrite oxidization reactions were performed when C/N = 0, but nitrite oxidization and partial denitrification occurred with low C/N ratios (0.44 or 0.88). At high C/N ratios (4.41 or 8.82), we mainly observed denitrification. In contrast to C/N = 0, the nitrite oxidization rate was unaffected when C/N = 0.44, but decreased with C/N = 0.88. The structure of bacterial communities varied significantly between autotrophic and mixotrophic conditions. Nitrobacter was hard to detect by PCR-DGGE while heterotrophs and especially denitrifiers were in the majority under mixotrophic conditions. Real-time PCR indicated that the Nitrobacter population decreased from 2.42 × 104 to 1.34 × 103 16S rRNA gene copies/ ng DNA, while the quantity of denitrifiers obviously increased from 0 to 2.51×104 nosZ gene copies/ ng DNA with an increasing C/N ratio. SDS-PAGE indicated the complexity of and a certain difference between the proteome of nitrite-oxidizing and heterotrophic bacteria at different C/N ratios. We conclude that the influence of organic matter on nitrite oxidation and the community structure of NOB and heterotrophic bacteria is complex. In this dissertation, we focused on how sodium acetate influenced the system both under suspended state and in biofilms. We observed that acetate did not necessarily have a negative impact on nitrification. Instead, an appropriate amount of acetate benefited both nitrite oxidization and denitrification. These findings provide a greater understanding about the relationship between organics and nitrification; they fill the gaps in the field of microbial ecology of nitrifying bacteria; they also provide insight into how to minimize the negative impact of heterotrophic bacteria and maximize the benefit of nitrogen removal in biological treatment systems.

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Silica-based functionalized terbium fluorescent nanoparticles were prepared, characterized and developed as a fluorescence probe for antibody labeling and time-resolved fluoroimmunoassay. The nanoparticles were prepared in a water-in-oil (W/O) microemulsion containing a strongly fluorescent Tb3+ chelate. N,N.N-1,N-1-12,6-bis(3'-aminomethyl-1'-pyrazolyl)phenylpyridine] tetrakis(acetate)-Tb3+ (BPTA-Tb3+), Triton X-100, octanol, and cyclohexane by controlling copolymerization of tetraethyl orthosilicate (TEOS) and 3-[2-(2- aminoethylamino)-ethylamino]propyl-trimethoxysilane (AEPS) with ammonia water. The characterizations by transmission electron microscopy and fluorometric quantum methods show that the nanoparticles are spherical and uniform in size, 45 +/- 3 nm in diameter, strongly fluorescent with fluorescence yield of 10% and a long fluorescence lifetime of 2.0 ms. The amino groups directly introduced to the nanoparticle's surface by using AEPS in the preparation made the surface modification and bioconjugation of the nanoparticles easier. The nanoparticle-labeled anti-human alpha-fetoprotein antibody was prepared and used for time-resolved fluoroimmunoassay of (x-fetoprotein (AFP) in human serum samples. The assay response is linear from 0.10 ng ml(-1) to about 100 ng ml(-1) with the detection limit of 0.10 ng ml(-1). The coefficient variations (CVs) of the method are less than 9.0%. and the recoveries are in the range of 84-98% for human serum sample measurements. (C) 2004 Elsevier B.V. All rights reserved.

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By using the home-made femtosecond laser system and the time-of-flight mass spectrometer, the decay dynamics of excited carbon disulfide (CS2) and ammonia (NH3) are investigated in real time by pump-probe multiphoton ionization detection. The estimated lifetime constant of the NH3 (A) over tilde (1)A(2)' state (51+/-4 fs) agreed quite well with the literature report. For the first time, the decay lifetime constants of the NH3 (E) over tilde'(1)A(1)' state (937+/-93 fs), the CS2 (a) over tilde (3)A(2) state (153+/-10 fs), and the CS2 Rydberg state [(3)/(2)]6ssigma(g) ((3)Pi(g)) (948+/-23 fs) are obtained.

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通过对黄土高原半湿润农田生态系统25年的田间肥料定位试验,研究了长期不同施肥模式对土壤有机氮组分及其在各级团聚体中分布的影响.结果表明:长期施肥对水解氨态氮、水解未知氮在土壤各级团聚体中分布的影响最大,对氨基酸态氮的分布有一定影响,而对氨基糖态氮分布的影响较小.长期施用化肥和有机肥能有效地影响水解氨态氮和水解未知氮与团聚体的结合作用,而氨基糖态氮在土壤氮循环转化过程中具有较强的稳定性.长期施肥条件下土壤水解全氮与有机碳、全氮以及团聚体分形维数均呈极显著正相关,其r分别为0.942,0.981,0.910(P<0.001),说明土壤有机氮组分对土壤团聚体的形成和性质具有显著影响.相关分析表明,土壤全氮或有机质对1~2mm和0.25~1mm土壤团聚体中各有机氮组分的影响较大

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利用不同坡度径流小区 ,在自然降水条件下 ,研究侵蚀泥沙不同形态有机氮的流失规律 ,结果表明 :在 4次产流泥沙中 ,除氨基糖氮含量减少外 ,其它有机形态氮均存在增加趋势 ,以氨态氮含量增加最为显著。随坡度的下降 ,水解全氮、氨态氮和氨基酸氮含量呈增加趋势 ;泥沙中水解全氮、氨基酸氮、氨态氮、非酸水解氮均存在不同程度的富集 ,其富集率随坡度的下降而增大 ,以氨基酸氮富集程度高 ,泥沙中氨基糖态氮反而无富集现象存在 ;当坡度分别为 2 5°、2 0°、15°、10°和 5°时 ,水解全氮流失量分别为 1135 .0、76 3.7、485 .5、313.0和 15 4.4kg/ km2 ,而氨基酸氮流失量则为 2 34.8、182 .7、12 1.1、76 .8和 33.9kg/ km2 ,占水解全氮的 2 0 .6 %~ 2 4.9%。氨态氮流失量为 742 .5、5 0 3.3、32 2 .2、2 0 8.9、97.4kg/ km2 ,占水解全氮的 6 3.1%~ 6 6 .7%。氨基糖氮占酸水解氮 1.1%~ 2 .9%。非鉴别氮占水解全氮的 5 .8%~ 12 .6 %。而酸水解氮与非酸水解氮之比分别为 ...

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Hydrogen sulfide (H2S) production patterns and the influence of oxygen (O-2) concentration were studied based on a well operated composting plant. A real-time, online multi-gas detection system was applied to monitor the concentrations of H2S and O-2 in the pile during composting. The results indicate that H2S was mainly produced during the early stage of composting, especially during the first 40 h. Lack of available O-2 was the main reason for H2S production. Maintaining the O-2 concentration higher than 14% in the pile could reduce H2S production. This study suggests that shortening the interval between aeration or aerating continuously to maintain a high O-2 concentration in the pile was an effective strategy for restraining H2S production in sewage sludge composting. (C) 2010 Elsevier Ltd. All rights reserved.

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In this paper, taking the northern region of Changxing County for example, with ammonia nitrogen as a pollution assessment index, we used an improved export coefficient method for estimate polluting load of non-point source pollution (NSP) and the social pollution survey data in the study area to estimate point source pollution. By comparing the total pollution output and the national surface water environmental quality standards find that the whole study area achieves the second water quality standard. However, Jiapu Township exceeds the water quality standards seriously because of the superfluous point source pollution. The water quality of other Townships is good. Further analysis showed that different types of land use and proportions in the northern region of Changxing County have a significant impact on the non-point source pollution, the general law is farmland contributes the largest share of the non-point source pollution output, followed by residential area and bare land, besides, with the increase in the proportion of forest and the decrease of farmland and residential area, the non-point source pollution reduces gradually. © 2010 IEEE.

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Nearly monodisperse and well-defined one-dimensional (1D) Gd2O3:Eu3+ nanorods and microrods were successfully prepared through a large-scale and facile hydrothermal method followed by a subsequent heat treatment process, without using any catalyst or template. X-ray diffraction (XRD), thermogravimetric analysis and differential scanning calorimetry (TGA-DSC), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED), photoluminescence (PL) and cathodoluminescence (CL) spectra as well as kinetic decays were used to characterize the samples. The size of the Gd2O3:Eu3+ rods could be modulated from micro- to nanoscale with the increase of pH value using ammonia solution. The as-formed product via the hydrothermal process, Gd(OH)(3):Eu3+, could transform to cubic Gd2O3:Eu3+ with the same morphology and a slight shrinking in size after a postannealing process.

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Uniform Gd(OH)(3) nanotubes have been prepared via a simple wet-chemical route at ambient pressure and low temperature, without any catalysts, templates, or substrates, in which Gd(NO3)(3) was used as the gallium source and ammonia as the alkali. SEM and TEM images indicate that the as-obtained Gd(OH)3 entirely consists of uniform nanotubes in high yield with diameters of about 40 nm and lengths of 200-300 nm. The temperature-dependent morphological evolution and the formation mechanism of the Gd(OH)(3) nanotubes were investigated in detail. Furthermore, the Gd2O3 and Eu3+-doped Gd2O3 nanotubes, which inherit their parents' morphology, were obtained during a direct annealing process in air. The corresponding Gd2O3:Eu3+ nanotubes exhibit the strong red emission corresponding to the D-5(0)-F-7(2), transition of the Eu3+ ions under UV light or low-voltage electron beam excitation, which might find potential applications in the fields such as light-emitting phosphors, advanced flat panel displays, or biological labeling.

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Uniform Lu2O3:Eu3+ nanorods and nanowires have been successfully prepared through a simple solution-based hydrothermal process followed by a subsequent calcination process without using any surfactant, catalyst, or template. On the basis of X-ray diffraction, thermogravimetric analysis and differential scanning calorimetry, and Fourier transform infrared spectroscopy results, it can be assumed that the as-obtained precursors have the structure formula of Lu4O(OH)(9)(NO3), which is a new phase and has not been reported. The morphology of the precursors could be modulated from nanorods to nanowires with the increase of pH value using ammonia solution. The as-formed precursors could transform to cubic Lu2O3:Eu3+ with the same morphology and a slight shrinkage in size after an annealing process, Both the Lu2O3:Eu3+ nanorods and nanowires exhibit the strong red emission corresponding to the D-5(0)-F-7(2) transition of the Eu3+ ions under UV light excitation or low-voltage electron beam excitation.