基于高通量测序的SBR反应器丝状膨胀污泥菌群分析

为探究丝状污泥膨胀及控制过程中细菌菌群和真菌菌群的变化规律,采用一套11 L的SBR反应器,接种某城市污水处理厂膨胀污泥,以乙酸钠为碳源进行人工配水,对膨胀污泥进行399 d的培养实验,采用高通量测序技术对污泥样本进行菌群多样性分析.结果表明,接种污泥菌群多样性比较丰富,经培养至污泥高度膨胀后,菌群多样性降低,污泥沉降性能恢复正常后,菌群多样性又逐渐增加.细菌中的腐螺旋菌属(Saprospiraceae_norank)、丛毛单胞菌属(Comamonadaceae_unclassified)和四球菌属(Tetrasphaera)相对丰度分别为13.37%、10.54%和8.59%,是接种污泥的主要细菌属.经过培养,膨胀污泥菌群发生变化,接种污泥中细菌相对丰度仅为0.01%的丝硫菌属(Thiothrix)增加至56.95%~60.14%,真菌中相对丰度为19.60%的丝孢菌属(Trichosporon)增加至94.82%.污泥膨胀得到控制后,污泥中丝硫菌属(Thiothrix)相对丰度减少至0.01%,丝孢菌属(Trichosporon)相对丰度减少至2.32%.丝硫菌属(Thiothrix)和丝孢菌属(Trichosporon)过多不利于污泥沉降.     

丝状菌污泥膨胀对脱氮除磷功能菌群的影响

丝状菌引起的污泥膨胀或生物泡沫是活性污泥法污水处理厂运行管理中经常碰到的异常问题,为确定丝状菌引起的污泥膨胀对脱氮除磷系统功能菌群的影响,采用形态学鉴定和Illumina MiSeq高通量测序对5座城市污水处理厂非膨胀期和膨胀期活性污泥、生物泡沫中关键微生物菌群分布特征进行分析研究.结果表明,污泥膨胀和生物泡沫主要由微丝菌(Microthrix parvicella)引起,膨胀期污泥和生物泡沫中微丝菌最高比例达6%和38%.主要脱氮除磷菌属为亚硝化单胞菌(Nitrosomonas)、硝化螺菌(Nitrospira)、陶厄氏菌(Thauera)和Candidatus Accumulibacter phosphatis.膨胀期与非膨胀期相比AOB和聚磷菌相对丰度明显降低,最大变化比例为54%和47%,反硝化菌相对丰度显著升高,最大变化比例为73%;脱氮除磷菌群的波动变化受污泥膨胀的影响外,还与处理工艺及菌群的生理特性相关.     

温度冲击引起的丝状菌污泥膨胀菌群特征

为探究温度冲击引起的污泥膨胀机理,以生活污水为处理对象,采用SBR工艺分别运行温度骤降系统和温度梯度降低系统,利用Illumina MiSeq高通量测序技术分析温度变化过程中微生物群落整体变化,并对膨胀阶段优势丝状菌类型进行解析.结果表明,温度骤降系统优势丝状菌为微丝菌(Microthrixparvicella),SVI值升高至291mL/g以上,温度梯度降低系统优势丝状菌为Eikelboom Type 0092型丝状菌,SVI值稳定维持在250mL/g,因此Eikelboom Type 0092型丝状菌适宜在温度冲击环境中生长繁殖.温度冲击方式不同导致菌群组成具有差异性,Proteobacteria相对丰度均值为39.3%,其占比在不同阶段变化较小.两个系统在污泥膨胀阶段Actinobacteria和Chloroflexi的相对丰度占比不同.各样本中与去除有机物相关微生物菌群丰度均值为13.6%,Nitrospira其相对丰度均值为2.48%,占NOB总含量80%以上.温度梯度降低系统发生的Eikelboom Type 0092型丝状菌型污泥微膨胀,其出水水质没有发生严重恶化,COD和NH₄⁺-N的去除效果均高于温度骤降系统.     

高钙废水颗粒污泥中古菌菌群结构变化的分析

为了解高浓度Ca~(2+)与颗粒污泥中古菌菌群结构的关系,利用高通量测序分析IC(Internal Circulation)反应器(以废纸造纸废水为处理对象)内不同高度处颗粒污泥中古菌菌群组成,并利用SEM(Scanning Electron Microscope)和EDS(Energy Dispersive X-ray Spectroscopy)分析颗粒污泥的形态结构和元素组成,同时借助红外光谱和XRD(X-ray diffraction)分析不同高度处颗粒污泥化学结构中各官能团的分布情况和无机成分.结果表明,随着反应器高度的增加,颗粒污泥中的无机成分CaCO_3也随着增加,而种泥中钙含量只有1.81%.种泥和ICR反应器内的颗粒污泥中古菌在门分类水平上主要包括Euryarchaeota和Bathyarchaeota,两者之和占90%以上.在属分类水平上,主要包括Methanosaeta、Methanoregula、Methanobacterium、Methanosarcina、Methanolinea、Methanospirillum等.种泥中古菌的物种多样性要明显高于IC反应器中,在IC反应器中不同高度处古菌生物多样性由高到低的顺序是5 m2 m7 m,5 m处的古菌物种更加丰富.随着反应器高度的增加,乙酸营养型甲烷菌的相对丰度呈现递增趋势,氢营养型甲烷菌则呈现递减趋势,而种泥中氢营养型甲烷菌相对丰度较大.这表明颗粒污泥在高钙废水的长期作用下,氢营养型甲烷菌的相对丰度减少,乙酸营养型甲烷菌的相对丰度增多,系统中古菌物种的相对丰度和多样性有所下降.CaCO_3沉淀使颗粒污泥的产甲烷活性降低,主要表现为其对氢营养型甲烷菌具有较大的抑制作用.     

低温时污泥膨胀对MBR中膜污染的影响

通过一体式膜生物反应装置考察了在低温条件下发生污泥膨胀过程中反应器的运行效果和膜污染的情况,并从微生物角度分析了引起膜污染的因素.结果表明,低温时COD上清液和出水平均去除率分别为85%和92%,发生丝状菌污泥膨胀后去除率变化不大.MBR中丝状菌污泥膨胀形成的过程中,污泥沉降性变差,丝状菌丰度（FI）由2增加到5,丝状菌伸出絮体形成网状结构.低温时膜操作压力随时间呈直线变化,膜组件的水力清洗周期为15 d.在丝状菌大量繁殖时缩短到7 d,膜污染严重.通过测定活性污泥的特性,发现膨胀污泥的胞外聚合物（EPS）总量是正常污泥的3倍,污泥絮体相对疏水性（RH）随FI的提高而增大.EPS和RH增大后会引起更多物质沉积到膜表面,使膜污染速率提高,膜的运行周期变短.进一步的分析表明,混合液粘度、Zeta电位、污泥絮体形态也是影响膜污染的因素.     

丝状菌膨胀污泥好氧颗粒化稳定性及微生物多样性

丝状菌作为污水生物处理中的常见菌种,被认为是诱发污泥膨胀的主要因素.然而由于其特殊的丝状形态,对污泥颗粒的形成起到了至关重要的作用.以丝状菌膨胀污泥为研究对象,探究丝状菌对污泥颗粒化过程及维持污泥颗粒稳定性的影响,并对污泥系统微生物多样性进行分析.试验分别接种丝状菌膨胀污泥(SVI=241.56 mL·g^-1)和絮状污泥(SVI=64.22 mL·g^-1)进行颗粒化培养,结果表明,膨胀污泥和絮状污泥颗粒出现时间分别为20 d和40 d,成熟颗粒粒径分别为650μm和700μm,膨胀污泥颗粒化时间仅为絮状污泥的一半.缺氧区添加后,颗粒都有不同程度破碎,但膨胀污泥培养的成熟颗粒SV₃₀/SV₅值短期波动后恢复至1,维持稳定性能力更强.微生物群落结构分析表明,膨胀污泥中norank＿o＿＿Saccharimonadales、unclassified＿o＿＿Saccharimonadales和unclassified＿f＿＿Saccharimonadaceae相对丰度从0.05%、0.01%和0.01%增长...     

FNA对短程硝化污泥菌群结构的影响

在SBR反应器增加游离亚硝酸（FNA）预处理单元,投加浓度为1.2mgHNO₂-N/L的FNA进行缺氧搅拌4.5h,连续处理3d,考察短程硝化污泥中FNA对氨氧化菌（AOB）,丝状菌和微生物菌群结构的影响.研究表明,FNA对AOB有短时抑制作用,并能够抑制优势丝状菌Candidatus_Microthrix（微丝菌属）和Cytophagaceae（噬纤维菌）的增殖,分别由5.1%和1.1%下降到0.78%和几乎不可见.SVI从281mL/g降低到100mL/g左右.NAR能够维持在90%左右,短程硝化不受到破坏.高通量结果显示,FNA处理后微生物菌群结构多样性与丰度出现下降,但Thauera（陶厄氏菌属）和Ottowia出现了增殖,分别增加到5.58%和7.82%,同步硝化反硝化（SND）作用明显,这使得即便只有短程硝化,总氮去除率依然能达到60%以上.     

低溶解氧污泥微膨胀的发生及脱氮除磷效果优化

采用SBR工艺以缺氧/好氧运行模式处理实际生活污水,在常温条件下通过降低溶解氧诱发活性污泥微膨胀,使污泥的容积指数(SVI)维持在150～220mL/g之间.研究了低溶解氧污泥微膨胀现象的发生和诱因微生物的生长,并对污泥微膨胀状态下系统的脱氮除磷效果进行优化.结果表明,常温条件下诱发污泥微膨胀的丝状菌主要有M.parcicella、0803型和H.hydrossis,其中M.parcicella为优势丝状菌,在丝状菌中所占比例最大.研究还发现,在低氧运行期间SBR出水正磷酸盐明显降低,COD略有升高但影响不大,但氨氮去除率明显下降,由95%以上下降到65%.通过采用厌氧/好氧/缺氧的运行模式:瞬间进水→厌氧搅拌(0.75h)→曝气(5h)→缺氧搅拌(2.25h)→静沉、排水(1h),氨氮和总氮的去除率均达到95%以上,出水总氮5mg/L,同时,PO43--P去除率达到95%以上,实现了低溶解氧污泥微膨胀系统高的脱氮除磷效率.     

XH02菌强化反应器脱氮过程中菌群结构的高通量分析

为了强化生物反应器脱氮以及揭示微生物菌群结构随时间的动态变化,以Ochrobactrum anthropi XH02和SBR反应器为研究对象,利用16Sr DNA高通量测序技术,对不同时期反应器中微生物的菌群结构和多样性进行了动态分析.研究结果表明,XH02菌的加入使得反应器中TN和COD的去除率分别提升了15%和10%以上;反应器中微生物菌群在属水平上的相对丰度和多样性呈先下降后上升的趋势;XH02的加入对菌群结构产生了较大影响.Acinetobacter、Blvii28和Aquabactenium菌的相对丰度显著下降,而Fontibacter和Treponema菌的相对丰度则在强化过程中显著升高;XH02的相对丰度随着反应器的运行逐渐增加,最后形成了较为稳定的菌群;主成分分析和UPGMA聚类分析大致把反应器运行过程分成4个阶段.     

干旱寒冷地区氧化沟工艺活性污泥的菌群结构研究

为了探讨新疆干旱寒冷地区城市污水厂活性污泥的菌群结构特征,采用高通量测序技术对北疆5座城市污水厂活性污泥进行菌群分析。结果表明:5座污水厂基本可以达到各自的设计出水标准,其中3座污水厂运行中出现了污泥丝状膨胀。变形菌门（Proteobacteria）、拟杆菌门（Bacteroidetes）和绿弯菌门（Chloroflexi）是活性污泥的优势细菌门,相对丰度分别为30.65%~43.12%、21.15%~33.64%和5.02%~14.36%。放线菌门（Actinobacteria）在沉降性能良好的污泥中相对丰度为2.97%~6.17%,而在丝状膨胀污泥中相对丰度达到11.69%~15.64%。季节性膨胀污泥中微丝菌属（Microthrix）和黄杆菌属（Flavobacterium）相对丰度呈现出季节性变化,常年膨胀的污泥中腐螺旋菌属（Saprospiraceae_norank）为优势细菌属,相对丰度达到17.48%~18.30%。子囊菌门（Ascomycota）是丝状污泥膨胀中的优势真菌门,相对丰度达到56.60%~72.68%。研究结果可对干旱寒冷地区污水厂运行调控提供理论支撑。     

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.     

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.     

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

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.