从不同反应器筛选、鉴别好氧反硝化菌

采用2个序批式反应器A和B,以硝态氮为唯一氮源,采用间歇曝气,以驯化、富集耐氧脱氮污泥.反应器A,其pH约为6.3,ρ(DO)为2.2～6.1 mg/L,碳氮比(ρ(C)/ρ(N),ρ(C)以ρ(COD_Cr)计)约为9;反应器B,其pH约为6.8～7.8,ρ(DO)为 2.2～3.0 mg/L,ρ(C)/ρ(N)约为15.2个反应器的ρ(NO₃－-N)均保持为80 mg/L.当2个反应器的总氮去除率达到60%以上,则认为完成好氧反硝化菌的富集.从2个反应器中共筛选得到20株BTB阳性菌,其中8株菌株的DNA样品经PCR成功扩增,进行16S rRNA测序.测序结果提交GenBank进行Blast同源性检索,并分析比对鉴别,判断8株菌株分属于假单胞菌(Pseudomonas),戴尔福特菌(Delftia),草螺菌(Herbaspirillum)和丛毛单胞菌(Comamonas)菌属.反硝化性能测定证实8株菌株均为好氧反硝化菌.      

好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究

采用污泥驯化手段富集好氧反硝化细菌,将得到的驯化污泥分离纯化,共得到5株好氧反硝化细菌.f1、f2、f3、f5和f7的TN去除率为90.4%、 91.2%、94.6%、95.6%和97%,表现出较好地去除总氮的能力.经过生理生化鉴定和16S rDNA测序, 建立了系统发育树,可基本确定分离的菌株f1、f3和f5为Pseudomonas sp., 分离菌f2和f4为Paracoccus sp..采用筛选的5株好氧反硝化细菌建立连续流反应器.在反应器进入稳定运行阶段时,可以观察到系统对于硝酸盐的去除率稳定在98.16%左右.表现出较好的硝酸盐去除效果,出水亚硝酸盐含量一直维持在较低的水平,其最大值不超过3.56　mg/L.COD的平均去除率为87.24％,基本实现了同一反应器中有机物和硝酸盐的共同去除.     

1株好氧反硝化菌的分离鉴定和反硝化特性研究

从沈阳北部污水处理厂曝气池的回流污泥中驯化分离得到16株有好氧反硝化能力的菌株,并最终筛选得到1株好氧反硝化能力较强的菌株N6。菌株N6的革兰氏染色为阴性、无芽孢;经16Sr DNA序列分析,鉴定其为假单胞菌(Pseudomonas sp).。反硝化特性实验表明:菌株反硝化的最佳温度是30℃、最适pH值为7、最佳C/N比为15∶1;碳源的种类对菌株的反硝化效果影响很大,菌株N6对丁二酸钠和乙酸钠等小分子碳源的利用相对高于对葡萄糖、蔗糖等大分子碳源的利用,菌株反硝化的最适碳源是丁二酸钠。在最佳降解条件下,菌株24 h对硝酸盐的降解率达98%,并且没有亚硝酸盐的积累。     

土著PHA合成菌回注法提高活性污泥积累聚羟基脂肪酸酯能力

在活性污泥驯化之初加入少量土著聚羟基脂肪酸酯(PHA)合成菌(0.10～0.25 g/L),通过10 d厌氧-好氧或好氧-沉淀方式进行污泥驯化后,以葡萄糖或乙酸钠为碳源进行PHA发酵.研究了驯化前后土著PHA合成菌回注组和对照组驯化过程中的总菌数,PHA合成菌数的变化,以及对PHA发酵产率的影响.结果表明:在回注组污泥中PHA合成菌的增长速率和增长量均高于对照组,各种污泥积累PHA占挥发性悬浮固体的比例提高了21.44%～43.18%.      

A2N双污泥反硝化除磷系统微生物的先间歇后连续培养及快速启动

以实际生活污水为对象,研究了反硝化聚磷菌(DPB)的驯化培养以及A2N双污泥反硝化除磷系统的快速启动.采用先独立培养反硝化聚磷菌和好氧硝化生物膜再连续运行的方式成功地快速启动了A2N系统.采用污水处理厂除磷工艺中的活性污泥为种泥,在SBR系统中以先A/O(厌氧,好氧)后A/A(厌氧,缺氧)的方式运行.32 d成功地使反硝化聚磷菌成为优势菌属.在SBR反应器中,采用硝化效果较好的活性污泥为种泥,好氧硝化生物膜30 d挂膜成功.氨氮去除率稳定在99%以上.然后.A2N系统连续运行,11d后系统反硝化除磷效果进入稳定状态,出水氨氮和正磷酸盐浓度均为O,硝态氮为10.26 mg/L,出水COD为19.56 mg/L,COD、氨氮、总氮和磷去除率分别为91%、100%、77%和100%,说明A:N系统具有很好的脱氮除磷效果,认为系统启动成功.     

异养硝化-好氧反硝化菌HY3-2的分离及脱氮特性

从经过高盐驯化的好氧颗粒污泥系统中筛选出一株异养硝化-好氧反硝化菌HY3-2,通过形态学观察及16S rDNA序列分析得出HY3-2为Klebsiella quasipneumoniae subsp.quasipneumoniae.研究了HY3-2对氨氮、硝酸盐和亚硝酸盐的去除特性,结果表明该菌具有良好的异养硝化和好氧反硝化功能,对氨氮、硝酸盐和亚硝酸盐的去除率分别达63.57%、88.11%和98.38%.对菌株脱氮性能研究表明：HY3-2以甘油为碳源,C/N为25,温度为20℃或30℃,转速为150r/min,盐度低于50g/L时,对100mg/L的NH₄⁺-N去除效果良好,去除率达90.7%;以柠檬酸钠为碳源,C/N为25,温度为30℃,转速为150r/min,盐度低于15g/L时能进行良好的好氧反硝化作用,NO₃^--N去除率达99%以上.     

活性污泥中好氧反硝化菌的富集筛选及鉴别

采用SBR反应器,以硝基氮为底物,通过间歇曝气方式,DO保持5mg/L以上,对活性污泥进行强化驯化,实现好氧反硝化细菌的富集培养。利用琥珀酸钠作为碳源,溴百里酚蓝(BTB)作为pH指示剂,共筛选得到20株BTB琼脂平板阳性菌。通过反硝化性能测定,复筛得到4株好氧反硝化细菌。实验结果表明,琥珀酸盐为碳源、硝酸盐为惟一氮源、C/N<10的条件下,4株菌在4d内的TIN去除率均达到60%以上。通过16SrRNA序列同源性比较成功鉴定出3株菌,初步判断2株属于Pseudomonas菌属、1株属于Delftia菌属。     

好氧反硝化菌的脱氮特性研究与探讨

本论文对好氧反硝化菌株FR2,FX5,FW4在好氧条件下的反硝化特性进行了研究。结果表明：反硝化主要发生在菌体的对数生长期,且反硝化过程中NO2^--N有少量的积累。 对不同温度、pH、碳源对好氧反硝化作用的影响进行了研究,结果表明：该三株菌在好氧条件下均具有良好的反硝化效果;温度为25℃～30℃的中性和微碱性条件反硝化效果较好;以乙酸钠为碳源的反硝化效果最好,NO3^-N的去除率最高。以葡萄糖为碳源的反硝化效果最差,NO3^-N的去除率最低。     

贫营养好氧反硝化菌的分离鉴定及其脱氮特性

为了分离贫营养好氧反硝化菌,研究其系统发育地位和脱氮特性,以期为微污染水库水体生物修复提供依据.从水库底层沉积物中,采用改良的富集驯化方法分离好氧反硝化菌,通过N-J法进行系统发育分析以及选择培养基研究其脱氮特性.初筛分离出196株好氧反硝化菌,其中14株为高效菌株(ZHF2、ZHF3、ZHF5、ZHF6、ZHF8、ZMF2、ZMF5、ZMF6、N299、G107、81Y、SF9、SF18和SXF14).经形态学,生理生化,和16S rRNA基因序列分析,ZHF3、ZHF5、ZHF6、ZMF2、G107、81Y、SF18和SXF14为不动杆菌(Acinetobacter sp.),ZHF2和ZHF8为新鞘脂菌属(Novosphingobium sp.),ZMF5为水杆菌属(Aquabacterium sp.),ZMF6为鞘脂单胞菌属(Sphingomonas sp.),N299为动胶杆菌属(Zoogloea sp.),SF9为代尔夫特菌属(Delftia sp.).其中菌株G107和81Y的反硝化效果最好,72h的硝氮去除率达到98.88% 和99.44%.并以G107和81Y为代表进行一系列的脱氮实验,结果显示出良好的短程反硝化、硝化和源水脱氮能力.贫营养好氧反硝化菌的分离,补充和丰富了好氧反硝化菌的种类,其高效的脱氮特性为低氮微污染水体的生物修复提供了技术支撑.     

同步脱氮好氧颗粒污泥的特性及其反应过程

厌氧颗粒污泥经过驯化后,成为具有同步硝化与反硝化(SND)功能的好氧颗粒污泥.实验在2L反应器中进行,温度,pH值,溶解氧分别控制在25℃,pH7～8,3～4mg/L.在实验条件下,SND好氧颗粒污泥COD去除率90%,氨氮去除率100%,出水检测不出NO₂^--N和NO₃^--N.反应器中SND颗粒污泥粒径在2.0～2.7mm的占全部颗粒污泥的50%,SVI为15～30mL/gTSS;污泥所能承受的最大压力为23.236N;SND好氧颗粒污泥中挥发性固体为9.92mg/mL,占总固体的2/3.采用SND好氧颗粒污泥进行脱氮研究,反应6h后氨氮去除率达100%,废水中检测不到NO₂^--N,仅残留2mg/L的NO₃^--N.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

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.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

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.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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