不同好氧颗粒污泥中微生物群落结构特点

为了探讨活性污泥好氧颗粒化过程对微生物种群的影响、不同底物及不同颗粒化方法培养的好氧颗粒污泥中微生物群落结构的差异,以接种污泥、模拟废水好氧颗粒污泥和分别投加粉末活性炭和硅藻土的实际生活污水好氧颗粒污泥为研究对象,利用PCR-DGGE对比分析了接种污泥和好氧颗粒污泥中的微生物群落结构.结果表明:活性污泥好氧颗粒化过程会减少微生物种群多样性,影响颗粒污泥稳定性的细菌被淘汰,而聚磷菌、反硝化菌、难降解有机物降解菌等污水处理功能微生物都在颗粒化过程中得到保留.活性污泥好氧颗粒化过程中能够实现亚硝化细菌(AOB)一定程度的富集.与接种活性污泥相比,好氧颗粒污泥中AOB的多样性指数与均匀性指数均有提高.好氧颗粒污泥中的优势菌群主要分布于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和未培养菌(uncultured bacterium).其中AOB均属于β-Proteobacteria的亚硝化单胞菌属(Nitrosomonas).     

工业有机废水为共基质培养降解五氯酚厌氧颗粒污泥

研究了工业有机废水和五氯酚(PCP)共存条件下,改良UASB反应器中厌氧污泥的颗粒化过程.实验发现降解PCP厌氧颗粒污泥形成可分成污泥驯化、颗粒污泥出现与成熟3个阶段,污泥颗粒化过程中污泥量、污泥活性和厌氧消化3大类群细菌数量与生态分布发生特征性变化,所培养颗粒污泥具有脱氯降解PCP的活性,产甲烷优势菌为杆状或长丝状产甲烷丝菌,在生物结构上形成互营微菌落,但污泥颗粒无明显层次性微结构.作为共基质的有机废水其种类影响着颗粒化进程、颗粒污泥脱氯活性与产甲烷优势菌.     

两次污泥颗粒化过程中微生物群落的动态变化

利用序批式反应器(sequencing batch reactor,SBR)培养好氧颗粒污泥(aerobic granular sludge,AGS),在此期间发生了AGS破碎现象,后经培养,破碎污泥再次变为成熟的AGS.因此,采用Illumina Mi Seq PE300高通量测序技术研究了两次污泥颗粒化过程中微生物群落结构变化的差异,以期揭示有利于AGS形成的优势菌属;此外,利用实时定量聚合酶链式反应(quantitative polymerase chain reaction,q PCR)探究了两次污泥颗粒化过程中硝化微生物的动态变化.结果表明,在两次污泥颗粒化过程中,胞外蛋白质和胞外多糖的含量均增加;氨氧化古菌(ammonia oxidizing archaea,AOA)在第一次污泥颗粒化过程以及AGS成熟过程丰度增加,氨氧化细菌(ammonia oxidizing bacteria,AOB)虽然在第一次污泥颗粒化过程中丰度降低,但是在AGS培养过程中其丰度都一直高于AOA;微生物群落多样性随着AGS的形成而降低;变形菌门(Proteobacteria)相对丰度明显增加,分别增加了12.29%和5.90%;某些属于变形菌门的属其相对丰度也增加,其中,Candidatus Competibacter在两次污泥颗粒化过程中相对丰度增加最明显,并且在成熟的AGS中呈现很高的相对丰度,达到14.20%.总的来说,胞外蛋白质和胞外多糖含量的增加,可能促进了污泥颗粒化;AOA和AOB可能共同参与了AGS的氨氧化作用;Ca.Competibacter的富集可能有利于AGS的形成.     

聚苯乙烯微塑料对菌-藻颗粒污泥的影响机制研究

在成熟的好氧颗粒污泥(AGS)中接种小球藻,成功构建了菌-藻共生好氧颗粒污泥(ABGS)系统.开展了聚苯乙烯微塑料(PS-MPs)对ABGS系统的影响机制研究.研究表明:低浓度PS-MPs(1mg/L)对ABGS的生物量、藻类生长、沉降性能和污染物去除效率无明显影响.相较于ABGS对照组,高浓度P S-MPs(10mg/L和100mg/L)胁迫作用下ABGS的生物量、藻类生长、沉降性能受到了明显抑制,叶绿素α/MLSS比值降低了0.19mg/g,污泥平均粒径降低了200μm,颗粒污泥结构逐渐松散破碎,有机物、氮和磷等污染物的去除效率分别降低了4.83%、5.06%和4.11%.此外,PS-MPs抑制了ABGS胞外聚合物(EPS)的分泌,降低了EPS中色氨酸和芳香族蛋白质的含量.在微生物群落结构组成方面,PS-MPs提高了ABGS系统内物种多样性,但抑制了疣微菌门(Verrucomicrobiota)、蓝藻门(Cyanobacteria)、绿弯菌门(Chloroflexi)等功能细菌的生长,不利于维持系统的长期稳定性.     

强化造粒条件下颗粒污泥中EPS不同组分的空间分布特征

采用多重荧光染色方法分析强化造粒条件下(不同时间段投加PAC)好氧颗粒污泥形成过程中胞外聚合物(extracellular polymeric substances,EPS)各组分的空间分布特征。研究发现:强化造粒条件下污泥完全颗粒化进程与对照组相比,所需时间明显缩短,粒径更大,含水率、比重、强度更优,且颗粒结构更规则紧密。EPS不同组分空间分布分析表明:α-呋喃葡萄糖、α-甘露糖主要分布在颗粒外侧,蛋白质在整个颗粒污泥断面均有分布,PAC投加对它们的分布影响较小,但对β-D-呋喃葡萄糖、脂类以及死细胞和活细胞在颗粒污泥中的分布有明显的影响;其中在1~7 d投加PAC,死细胞和β-D-呋喃葡萄糖主要集中在颗粒污泥外侧,活细胞和脂类主要分布在颗粒污泥内部,而8~14 d投加PAC,上述情况刚好相反;对照组(不投加PAC)颗粒污泥EPS各组分的空间分布与推迟PAC投加时间后的情况基本相似。     

降解五氯酚(PCP)的微氧颗粒污泥的形成机理

在微氧条件及剧毒性的五氯酚存在下,研究了活性污泥的颗粒化机理.在污泥培养过程中对污泥性质、外观进行观察,并利用扫描电镜对微生物相进行分析.结果表明,在颗粒化初始阶段诱导核的存在对絮体起着凝集作用,该阶段的微生物以丝状菌为主;在形成阶段,二价金属阳离子含量以及ECPs中PN/PS比值增加,丝状菌形成三维框架结构;而在颗粒污泥形成后的增长阶段,SVI值下降至较低的水平,形成表面光滑的完整颗粒,颗粒内杆菌、球菌占优势,含水率的明显下降表明质子跨膜迁移引起的细胞脱水作用变得显著.     

以脱水污泥为接种污泥促进好氧污泥颗粒化

以脱水造粒形成的物理颗粒污泥为接种污泥,明显提高了好氧污泥颗粒化速度.研究结果表明：在第20d,接种物理颗粒污泥的R2中90%以上的污泥粒径即大于0.2mm,而接种絮状污泥的R1中只有26.7%.颗粒化过程中,接种物理颗粒污泥的R2中SVI始终小于80mL/g沉降性能良好,第25d时污泥浓度为6300mg/L,而R1为3200mg/L.脱水过程未对污泥活性造成明显影响,培养期间两者COD去除率均大于90%,但培养后期R2中TN的去除率约为70%,明显优于R1的55%,其主要原因为R2中的污泥粒径大于R1.经过5d的曝气剪切后仍有39.8%的物理颗粒污泥大于0.2mm,为颗粒化提供了大量诱导核,同时物理颗粒污泥内部营养传输孔道的形成与EPS的內源消化和反硝化产气有关.     

好氧颗粒污泥快速培养影响参数及方法研究进展

针对好氧颗粒污泥培养速度慢、启动周期长等突出问题,通过综述好氧颗粒污泥的形成机理、颗粒化主要影响参数以及促进颗粒化的方法,发现调整反应器的水力剪切力、沉降时间、有机负荷、饥饿期等参数有利于诱导微生物分泌更多的胞外聚合物(EPS),促进初期微生物聚集体的快速形成,从而缩短好氧颗粒化时间。因此,当前快速颗粒化的方法基本上是基于诱导初期颗粒聚集体快速形成或直接投加聚集体的方法缩短好氧颗粒化时。最后,总结好氧颗粒快速培养过程中存在问题,并提出好氧颗粒污泥形成机理的明晰、好氧颗粒化的标准以及培养指标体系的建立是解决相关问题的关键,以及今后的研究重点。     

好氧污泥颗粒化过程中Zeta电位与EPS的变化特性

研究了好氧颗粒污泥在形成过程中Zeta电位、胞外多聚物的变化以及两者之间的关系.通过对污泥调控生长,形成完全颗粒化的好氧污泥.在好氧颗粒形成过程中,污泥的Zeta电位由接种时的-19.1 mV逐渐降至-10.1 mV.完全颗粒化以后,成熟后的好氧颗粒污泥Zeta电位稳定维持在-10 mV左右,说明维持污泥较低的Zeta电位是好氧颗粒污泥形成的重要条件.颗粒化过程中污泥EPS中多糖含量基本不变,蛋白含量增加明显,从13.65 mg.g-1增加到54.12 mg.g-1,蛋白/多糖比值从0.67增加到3.31;污泥的Zeta电位与蛋白/多糖的比值呈正相关,相关系数为0.849,这些结果说明蛋白质对降低污泥表面电位,促进污泥聚集以及好氧颗粒污泥的形成生长等可能起着重要作用.     

同步脱氮除磷好氧颗粒污泥培养过程微生物群落变化

本实验利用生活污水培养具有同步脱氮除磷(simultaneous nitrogen and phosphorus removal,SNPR)功能的好氧颗粒污泥(aerobic granular sludge,AGS).采用Illumina Mi Seq PE300高通量测序对AGS培养过程中细菌群落变化进行了研究,以期揭示污泥好氧颗粒化成因.采用实时荧光定量PCR对AGS培养过程中氨氧化细菌(ammonia-oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)、亚硝酸盐氧化菌(nitrite-oxidizing bacteria,NOB)和聚磷菌(polyphosphate accumulating organisms,PAOs)的丰度变化进行了研究.结果表明:历时100 d培养出的AGS质地紧实,具有良好的SNPR效果.AGS胞外聚合物(extracellular polymeric substances,EPS)中多糖含量在培养过程中增加明显,而蛋白质含量保持稳定.AGS培养过程中,AOB的丰度略微下降,AOA的丰度明显下降,NOB的丰度无明显变化,而PAOs的丰度在AGS培养初期明显增加.在AGS培养过程中,细菌群落多样性呈现出先升高后降低的趋势,且细菌群落组成发生了明显的变化.持久型OTUs占样品总序列数的92.70%,其中变形菌门(Proteobacteria,31.07%~53.67%)、拟杆菌门(Bacteroidetes,6.70%~16.50%)和绿弯菌门(Chloroflexi,7.84%~13.36%)是AGS培养过程中的细菌优势门.Candidatus competibacter属在AGS培养过程中大量富集(由种泥中的0.11%增加到35.33%),其可能会分泌胞外多糖,形成黏性EPS,促进絮状污泥团聚成为AGS.     

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.     

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.     

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.     

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.     

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.     

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.     

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.