高效反硝化细菌的快速培养及群落结构多样性分析

培养高效的反硝化细菌可提高污水处理效率.本实验为序批式实验,以Ⅰ号、Ⅱ号发酵液为碳源,采用梯度提高硝氮的方式,培养高效的反硝化细菌,从中选择培养更加快速的发酵液.并采用高通量测序技术分析反硝化细菌的生物群落结构和多样性的变化.结果表明,Ⅱ号发酵液能够在第11 d便实现高效反硝化细菌[300 mg·(L·h)-1]的快速培养,比Ⅰ号发酵液提前了17 d,同时,高效反硝化细菌系统对氨氮和总磷有一定的去除效果,最大去除速率分别为34.43 mg·(L·h)-1和2.98mg·(L·h)-1.高通量测序分析结果表明,污泥经过驯化培养,物种丰度和多样性降低,但发挥反硝化作用的优势菌群的类别和比例得到增大;细菌的组成及数量发生了较大的改变,最终发挥高效反硝化作用的核心菌属为Thauera和Pseudomonas.另外,反硝化聚磷菌科(Rhodocyclaceae和Pseudomonadaceae)和异养硝化菌属(Pseudomonas、Alcaligenes、Bacillus和Comamonas)的存在,验证了系统对氨氮和总磷的去除能力.     

PCR-DGGE研究处理垃圾渗滤液序批式生物膜反应器(SBBR)中的细菌多样性

为了研究序批式生物膜反应器中的细菌多样性及其脱氮的微生物学机理,为工艺改进提供依据,从同步高效去除垃圾渗滤液中高氨氮和高COD的SBBR生物膜和渗滤液原水中采集微生物样品并提取微生物总DNA,使用细菌通用引物对（GC341F/907R）从总DNA中成功扩增出目标16S rDNA片段,然后对扩增的16S rDNA进行DGGE,对凝胶染色并进行条带统计分析和切胶测序,使用序列数据进行同源性分析并建立了系统发育树.结果表明,该驯化后的SBBR生物膜和渗滤液原水中都有比较丰富的细菌多样性,驯化的生物膜细菌主要来自渗滤液原水,而且生物膜细菌在反应器正常运行时不会出现明显的群落结构变化;在该SBBR中有多种硝化细菌与反硝化细菌、好氧反硝化细菌和厌氧氨氧化细菌共存,说明该反应器中可能同时存在全程硝化反硝化、同步硝化反硝化和厌氧氨氧化3种脱氮方式.研究结果为SBBR脱氮微生物机理研究提供了一些有价值的参考依据.     

序批式生物反应器填埋场脱氮微生物多样性分析

为探究序批式生物反应器填埋场脱氮过程中的微生物作用机制,本研究采用建立脱氮功能基因(amoA、nosZ)克隆文库及PCR-RFLP技术对序批式生物反应器填埋场垃圾稳定化后期的主要脱氮功能微生物多样性进行分析.结果表明,矿化垃圾反应器中检测到的氨氧化细菌存在高度多样性,大部分为未知类群,且均为不可培养菌或未经分离获得的细菌,经系统发育树分析系统内氨氧化细菌以β-变形菌门中的亚硝化单胞菌属(Nitrosomonas)为主;新鲜垃圾反应器中反硝化细菌种群丰富,主要有β-变形菌纲中的陶厄氏菌属(Thauera)和硫杆菌属(Thiobacillus).Thauera属在好氧条件下具有反硝化特性,Thiobacillus属中的脱氮硫杆菌(Thiobacillus denitrificans)是一种硫自养反硝化菌,可见新鲜垃圾单元稳定化后期以好氧反硝化和自养反硝化的脱氮途径为主.此外文库中检测到的一部分反硝化细菌可能归属于α-变形菌纲的慢生根瘤菌科(Bradyrhizobiaceae).     

矿化垃圾生物反应器中的细菌多样性分析

为探究准好氧生物矿化垃圾床处理渗滤液过程中的微生物作用机理,研究建立了16S r DNA克隆文库及PCRRFLP技术以研究矿化垃圾反应器的细菌多样性。结果表明:矿化垃圾反应器细菌具有高度多样性,反应器内有36种细菌分别属于13个纲,变形菌纲占绝对优势占70%(其中β-变形占56.5%,γ-变形菌纲占10%),拟杆菌纲、鞘氨醇菌纲,芽孢杆菌纲也具有一定优势;3%的Nitrosomonas属是渗滤液氨氮转化成亚硝酸盐氮的主要功能微生物,由于亚硝酸盐氧化菌不存在或丰度极低,因此造成反应器内亚硝酸盐积累;Thauera属(17%)和Thiobacillus denitrificans属(10%)是反应器内主要优势微生物属,是反应器内反硝化脱氮的功能微生物,由于Thauera属在好氧条件下具有反硝化特性,Thiobacillus denitrificans为严格自养反硝化菌,因此反应器脱氮主要途径为好氧反硝化、自养反硝化。     

硫自养反硝化与厌氧氨氧化耦合工艺中微生物群落结构和多样性研究

为研究基于硫自养反硝化和厌氧氨氧化耦合工艺中微生物群落结构和多样性特征,从稳定运行的HABR厌氧折流板反应器中采集生物膜样品,利用PCR-DGGE技术分别对反应器中4个格室进行群落结构解析。结果表明,微生物群落变化与水质环境中的有机物、氮和硫的浓度有关,微生物群落结构在不同的格室中变化较大,4个格室中的细菌的Shannon-Wiener指数(H)由左至右依次减小,相邻格室的相似性较高。经过克隆测序分析,HABR反应器内的微生物多为硫自养反硝化菌和厌氧氨氧化菌,并且出现了Sulfurimonas菌种,该类菌种是同步脱硫反硝化作用的主要功能菌,它们对污水中硫及氮的去除发挥了重要的作用。     

缓释碳源生态基质对低碳氮比河水脱氮效果研究

通过缓释碳源生态基质颗粒脱氮效果实验,比较了缓释碳源生态基质颗粒填料柱与普通砾石填料柱对各种形态氮的去除效果.结果发现,装填生态基质颗粒的实验组出水NO_2~--N、NO_3~--N和TN去除率分别为90.60%、90.32%和63.66%,明显高于对照组-16.39%、-1.51%和25.06%的去除率,说明缓释碳源生态基质可显著增强反硝化作用强度,提高TN去除率.高通量分析结果表明,生态基质组相对丰度超过1%的菌属数量高于对照组,其中,反硝化菌属相对丰度达到30%以上,生态基质释放的纤维素碳源有利于异养反硝化微生物的生长繁殖,使反应器内的微生物群落结构发生显著改变,提高了脱氮效率.     

高原湖泊沉积物中反硝化微生物的群落特征——以包头南海湖为例

为了探明内蒙古高原富营养化城市湖泊中nirS型反硝化细菌群落结构的特异性,利用特异性功能基因nirS对包头市南海湖的表层沉积物进行反硝化细菌多样性测定,在分析各采样点沉积物理化指标的基础上,对各样品中nirS型反硝化细菌群落和多样性进行研究.结果表明,南海湖沉积物Shannon指数变化为3.07~3.41,相比于其他研究,反硝化微生物多样性相对较低.参与反硝化的优势菌属与大多数湖泊保持一致,以假单胞菌属（Pseudomonas）和红杆菌属（Rhodobacter）为主.冗余分析（RDA）指出,城市湖泊中的各种类型的污染物对反硝化优势菌属有明显的促进作用,红杆菌属（Rhodobacter）与硝酸盐氮、亚硝酸盐氮呈正相关,假单胞菌属（Pseudomonas）除受硝酸盐氮、亚硝酸盐氮的影响外,还与总磷呈较强的正相关.南海湖nirS型反硝化细菌中的主要菌属促进了反硝化作用,加速了南海湖氮素的去除.     

凤眼莲对富营养化水体中氨氧化和反硝化微生物的影响

凤眼莲近年来广泛应用于富营养化淡水湖泊的生态修复中,但其对微生物的相互作用和对水体中氮素的去除鲜有报道.本研究在氮素去除过程中对比凤眼莲和细菌的相对重要性,并且检测浮游植物对硝化细菌和反硝化细菌相对丰度及多样性的影响.水体中氮素的去除率以及硝化和反硝化作用的潜在能力使用定量聚合酶链式反应(qPCRs)对硝化作用基因amoA和反硝化作用基因nirS/K进行检测,从微观角度研究富营养化水体中是否会受到凤眼莲存在的影响.结果表明,TN的减少在70d的实验周期中所有处理组表现较为一致,但凤眼莲存在的实验组在24h内TN和NH_4~+-N的去除上有显著的降低,并且amoA的丰度有所增加,nirS/K的丰度有所降低.T-RFLP结果表明亚硝化单胞菌在氨氧化微生物中占优势.凤眼莲的种植可以实现富营养化水体中NH_4~+-N的快速有效减少,且微生物的相互作用可以充分利用到淡水生态系统的修复中.     

河北平原潮土中微生物对氮降解特征

了解土壤中微生物对氮素降解规律,对于土壤氮污染修复具有重要现实意义.从河北平原潮土中筛选出8种高效氨化细菌、硝化细菌、异养硝化好氧反硝化细菌,对所筛菌种进行16S r DNA分析,选出最适菌株通过固定化载体将菌种制成菌剂.探讨了用加入所筛菌种及拮抗菌的菌液和地表水分别滴灌小白菜对土壤中的氮降解产生的影响,并测定土样磷脂脂肪酸(PLFA)的值.结果表明:以硅藻土为载体硝化细菌的氨氮降解率达到38%,硝态氮合成率为205%;以硅藻土为载体的氨化细菌氨氮合成率为1 711%,异养硝化好氧反硝化细菌的硝态氮降解率达到367%.加所筛菌液滴灌对土壤总氮和氨氮的降解效果较好,硝化作用也较强,加菌后土壤中微生物量明显增加,且更快到达峰值.对不同土样不同时间磷脂脂肪酸数据分析发现,在白菜生长过程中,土壤中的微生物含量均产生了1次峰值,且加入所筛菌的先到达峰值.对于任丘和阜城土样来说,加入所筛菌后,土壤中微生物种类相应增多,对土壤微生物环境有所改善,微生物也比较活跃.     

平原水库与地下水交互带中氨氮转化规律研究

为掌握年调节型平原水库与地下水交互带氨氮转化规律,以辽河石佛寺水库为例,通过采集交互带土样进行土柱渗水试验和微生物试验,分析氨氮的转化过程及特性,并结合一级反应动力学模型计算氨氮及产物转化速度常数。结果表明土柱0~30 cm在硝化细菌和亚硝化细菌作用下主要发生硝化过程;30~60 cm硝化和反硝化过程同时存在;60~80 cm在反硝化细菌作用下主要发生反硝化过程,到试验结束硝酸盐氮浓度接近0 mg/L。经计算,氨氮、亚硝酸盐、硝酸盐氮的转化速度常数分别为0.059、0.049、0.036 d-1。由此推断在平原型水库交互带内40 cm以上,氨氮转化以硝化作用为主;60 cm以下以反硝化作用为主,菌种分布规律与氨氮转化过程相一致。该研究为掌握平原型水库交互带氨氮转化机制和保护库区地下水环境提供科学依据。     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

基于综合水质标识指数和对应分析法的校园河道水质评价

以三峡大学的校园河道求索溪为研究对象,利用综合水质标识指数法确定求索溪水质类别,分析其水质时空变化规律,并利用对应分析法得出求索溪中不同监测点的主要污染因子.研究结果表明:求索溪整体的综合水质标识指数为7.423,整体水质为劣V类(地表水环境质量标准GB 3838-2002)且黑臭.从时间变化来看,求索溪4月份的水质最差,5月份次之,4、5月份所有监测点的水质都劣于V类且黑臭;8月份水质最好,水质为Ⅳ类;从空间分布来看,8个监测点综合水质标识指数均超过6.0,水质为劣V类,其中6号监测点的水质相对最好,监测点3号的水质相对最差;对应分析法得出求索溪的整体水体污染程度受总氮因子的影响最大,其次为总磷.该研究拟为求索溪及类似校园河道的水环境治理研究提供基础依据和参考.     

Effects of chitosan on growth of an aquatic plant (Hydrilla verticillata) in polluted waters with different chemical oxygen demands

Effects of chitosan on a submersed plant, Hydrilla verticillata, were investigated. Results indicated that H. venicillata could prevent ultrastructure phytotoxicities and oxidativereaction from polluted water with high chemical oxygen demand (COD). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in H. verticillata treated with 0.1% chitosan in wastewater increased with high COD (980 mg/L) and decreased with low COD (63 mg/L), respectively. Ultrastructural analysis showed that the stroma and grana of chloroplast basically remained normal. However, plant cells from the control experiment (untreated with chitosan) were vacuolated and the cell interval increased. The relict of protoplast moved to the center, with cells tending to disjoint. Our findings indicate that wastewater with high COD concentration can cause a substantial damage to submersed plant, nevertheless, chitosan probably could alleviate the membrane lipid peroxidization and ultrastructure phytotoxicities, and protect plant cells from stress of high COD concentration polluted water.     

Removal of cadmium using MnO2 loaded D301 resin

MnO2 loaded weak basic anion exchange resin D301 （Anion exchange resin, macroreticular weak basic styrene） as adsorbent has been prepared and applied to the removal of cadmium. The adsorption characteristics have been investigated with respect to effect of pH, equilibrium isotherms, removal kinetic data, and interference of the coexisting ions. The results indicated that the Cd^2＋ could be efficiently removed using MnO2 loaded D301 resin in the pH range of 3-8 from aqueous solutions with the co-existence of high concentration of alkali and alkaline-earth metals ions. The saturate adsorption capacity of the Cd^2＋ was 77.88 mg/g. The adsorption process followed the pseudo first-order kinetics. The equilibrium data obtained in this study accorded excellently with the Langmuir adsorption isotherm.     

Effect of coagulation pretreatment on the fouling of ultrafiltration membrane

The purpose of this study is to understand the effect and mechanism of preventing membrane fouling, by coagulation pretreatment, in terms of fractional component and molecular weight of natural organic matter （NOM）. A relatively higher molecular weight （MW） of hydrophobic compounds was responsible for a rapid decline in the ultrafiltration flux. Coagulation could effectively remove the hydrophobic organics, resulting in the increase of flux. It was found that a lower MW of neutral hydrophilic compounds, which could remove inadequately by coagulation, was responsible for the slow declining flux. The fluxes in the filtration of coagulated water and supernatant water were compared and the results showed that a lower MW of neutral hydrophilic compounds remained in the supernatant water after coagulation could be rejected by a membrane, resulting in fouling. It was also found that the coagulated flocs could absorb neutral hydrophilic compounds effectively. Therefore, with the coagulated flocs formed on the membrane surface, the flux decline could be improved.     

Penetration analysis of elements and bioleaching treatment of spent refractory for recycling

Acidithiobacillus ferrooxidans ATCC23270 and Acidithiobacillus thiooxidans TM-32 were used for bioleaching of spent refractories of aluminium and copper melting furnaces for their recycling.Firstly,penetration of elements into aluminium melting furnace refractory was investigated and it was found that up to 7 cm from surface was contaminated.Comparison on leaching efficiency by the strains ATCC23270 and TM-32 found that the strain ATCC23270 could treat larger amount of the refractories than the strain TM-32 could do.In the experiment of bioleaching of spent refractory aluminium melting furnace by the strain ATCC23270,high leaching efficiency were obtained on Al,Si,and Ca,and extremely low leaching performance was,however,shown on the rest of elements i.e.,Na,Mn,and Zn.Under the strain TM-32 use,relatively high leaching performance was recognized on Al,Si,Ca,Na,Mn,and Zn.In the experiment of bioleaching for spent refractory copper melting furnace,almost the same leaching trends were shown on Cu,Zn,Al,and Si under the strains ACTT23270 and TM-32 uses.     

Simultaneous removal of ammonium and phosphate by zeolite synthesized from coal fly ash as influenced by acid treatment

Zeolite synthesized from fly ash （ZFA） without modification is not efficient for the purification of NH4＋ and phosphate at low concentrations that occur in real effluents, despite the high potential removal capacity. To develop an effective technique to enhance the removal efficiency of ammonium and phosphate at low concentrations, ZFA was modified with acid treatment and the simultaneous removal of ammonium and phosphate in a wide range of concentration was investigated. It was seen that when compared with untreated ZFA, only the treatment by 0.01 mol/L of H2SO4 significantly improved the removal efficiency of ammonium at low initial concentrations. The behavior was well explained by the pH effect. Treatment by more concentrated H2SO4 led to the deterioration of the ZFA structure and a decrease in the cation exchange capacity. Treatment by 0.01 mol/L H2SO4 improved the removal efficiency of phosphate by ZFA at all initial P concentrations, while the treatment by concentrated H2SO4 （≥0.9 mol/L） resulted in a limited maximum phosphate immobilization capacity （PIC）. It was concluded that through a previous mild acid treatment （e.g. 0.01 mol/L of H2SO4）, ZFA can be used in the simultaneous removal of NH4＋ and P at low concentrations simulating real effluent.