MBBR-MBR组合工艺处理生活污水试验中微生物种群结构的演变

采用移动床生物膜反应器(MBBR)-膜生物反应器(MBR)组合工艺进行生活污水处理试验,考察了组合工艺对污染物的去除效果,通过变性梯度凝胶电泳(DGGE)分析反应器内微生物种群组成及多样性变化,利用克隆文库分析微生物种群结构。结果表明:MBBR-MBR组合工艺稳定运行时对氨氮($NH_{4}^{+}$-N)和化学需氧量(COD_Cr)的平均去除率高达98%和97%;DGGE指纹图谱显示,反应器运行过程中微生物群落结构演变明显且与温度变化密切相关,逐渐形成适应污水处理的低温稳定种群,MBBR与MBR相比,其反应器内微生物种群更丰富,抗冲击负荷能力更强;克隆文库结果显示,MBBR与MBR内微生物优势种有较大差异,但同属变形菌门(Proteobacteria)。     

胞外聚合物在生物膜同步去除/富集磷酸盐系统中的作用

以基于同步去除/富集磷酸盐的厌氧/好氧交替生物膜序批式反应器内生物膜为研究对象,研究了胞外聚合物（EPS）内磷含量形态及生物膜内微生物种群变化,探究EPS在生物膜去除/富集磷酸盐中的作用及其与微生物种群之间的联系.结果表明,生物膜反应器在厌氧外加COD为200 mg·L^-1的条件下富集到了磷浓度为120.95 mg·L^-1的富集液.EPS在生物膜吸/释磷过程中发挥重要作用,EPS磷含量占生物膜磷含量的69.16%~79.00%,³¹P核磁共振实验表明ortho-P为EPS内主要磷形态,占比为85.47%~88.60%.高通量测序结果表明生物膜内微生物种群变化明显,Candidatus_Competibacter为优势菌属,其丰度由1.23%增至38.87%,有利于形成更具粘性的EPS进而黏附在生物膜上,可能促进EPS在吸/释磷中发挥作用;暖绳菌科为优势聚磷菌,随实验温度升高,其丰度由5.29%减至4.90%.     

分格复合填料曝气生物滤池脱氮除磷特性及微生物群落特征分析

设计一种新型曝气生物滤池-分格复合填料曝气生物滤池,采用污水处理厂曝气沉砂池出水为初始微污染水,调试反应器对微污染水的深度处理效果,同时对填料表面生物膜中的微生物群落组成和结构进行解析.结果表明:该工艺在曝气量为3.0 L·min-1,水力负荷为20m3·m-3·d-1,回流比为125%的条件下,对化学需氧量(COD)、总氮(TN)、氨氮、浊度的平均去除率分别为81%、48%、91%、91%;该工艺对磷的去除率仅为14%,通过投加聚合氯化铝可使出水中总磷(TP)含量降至0.50 mg·L-1以下.微生物群落分析发现,好氧段中与硝化过程中密切相关的硝化螺菌属微生物占群落总量的8.0%,而在缺氧段与反硝化相关的脱氯单胞菌属微生物可占群落总量的1.5%,微生物群落组成与反应器功能具有较好的一致性.本研究表明,该工艺可以高效去除总氮,与聚合氯化铝联合作用可去除86%的总磷,达到污水排放一级A标准,可作为城市杂用水和景观环境用水进行回用.     

Ca2+、Mg2+共存对SBR工艺生物脱氮和微生物群落结构的影响

为了研究共存的硬度金属离子在废水生物处理中的作用,在进水Ca~(2+)为1.1 mmol·L~(-1)的条件下,通过改变Mg~(2+)的浓度,考察Ca~(2+)/Mg~(2+)物质的量比对SBR工艺污染物去除和微生物群落的影响,采用高通量测序技术分析微生物优势种群的变化,以期从微生物角度明确Ca~(2+)、Mg~(2+)共存对生物脱氮的影响机制.结果表明:当Ca~(2+)/Mg~(2+)物质的量比分别为2、1和0.5时,COD去除率由88%分别升高至90%、91%和93%;NH~+_4-N去除率由74%分别升高至91%、93%和96%;TN去除率由44%分别升高至58%、62%和69%.随着进水Ca~(2+)/Mg~(2+)物质的量比的降低,微生物群落的丰富度升高,Ca~(2+)/Mg~(2+)物质的量比为2的微生物群落结构与Ca~(2+)/Mg~(2+)物质的量比为1和0.5的微生物群落结构差异显著.变形菌门、拟杆菌门和放线菌门一直为SBR工艺的优势菌门,有利于有机污染物的去除.参与脱氮过程的Niabella和Dechloromonas在反应器内富集,保证了良好的脱氮效果.Ca~(2+)/Mg~(2+)物质的量比的降低促进了有机物和总氮的去除及微生物多样性的提高.     

膜曝气-生物膜反应器生物强化处理阿特拉津废水运行性能

在疏水SPG(shirasu porous glass)膜表面形成基因工程菌生物膜,构建SPG膜曝气-生物膜反应器(MABR)生物强化处理阿特拉津废水,考察MABR反应器稳定运行过程中污染物去除性能及其影响因素.结果表明,增大SPG膜孔径和曝气压力,能够提高曝气供氧能力,改善COD和阿特拉津生物强化去除效能.1.5μm疏水SPG膜在70 k Pa曝气压力下的最大供氧能力约为22.4 g·(m~2·d)~(-1).曝气压力为70 k Pa、水力停留时间(HRT)为1.5 h时,1.5μm膜MABR反应器COD平均去除率为80.1%,平均去除负荷为1.86 kg·(m~3·d)~(-1);阿特拉津平均去除率为62.5%,平均去除负荷为0.18 kg·(m~3·d)~(-1).进一步缩短HRT、增加进水负荷后,MABR反应器DO浓度显著下降,COD和阿特拉津去除效率大幅降低.DO浓度对阿特拉津去除的影响更为显著.随着MABR反应器的稳定运行,SPG膜表面单一基因工程菌生物膜逐渐演化为复杂微生物群落,但基因工程菌可以较好地存在于生物膜内,从而保持阿特拉津生物强化去除能力.     

SPG膜曝气-基因工程菌生物膜反应器处理阿特拉津废水研究

膜曝气-生物膜反应器(MABR)是一种新型的膜-生物废水处理工艺,在MABR中采用基因工程菌生物膜可以强化难降解污染物的生物去除.本研究在SPG膜表面形成基因工程菌生物膜,运行SPG膜曝气-生物膜反应器(SPG-MABR)处理阿特拉津废水,考察了气压、挂膜生物量和液体流速对SPG-MABR运行性能的影响,以及基因工程菌生物膜的变化.结果表明,提高气压可以增大透氧系数,从而提高阿特拉津和COD的去除速率以及复氧速率.提高挂膜生物量能够加快阿特拉津和COD的生物去除,但生物膜厚度增加使得氧传质阻力增大,复氧速率降低.层流状态下减小SPG-MABR中的液体流速,有利于污染物向生物膜扩散传质,从而提高污染物去除速率.气压为300 kPa、生物量为25 g·m-2、液体流速为0.05 m·s-1时,SPGMABR反应器对阿特拉津5 d的去除率可以达到98.6%.在SPG-MABR运行过程中,基因工程菌生物膜呈现微生物多态化趋势.生物膜表面逐渐被其他微生物细胞覆盖,基因工程菌分布减少,生物膜内部仍以基因工程菌细胞为主.     

HN-AD菌生物强化接触氧化工艺处理猪场沼液

针对养猪废水采用传统工艺经厌氧发酵处理后,形成高氨氮低碳氮比沼液,导致脱氮效果差、工艺流程复杂、启动周期较长等问题.本研究以异养硝化-好氧反硝化(HN-AD)菌为生物强化剂,以PAN活性炭纤维作为填料的生物接触氧化池(BCO)为生物膜反应器,形成生物强化BCO工艺处理猪场沼液.前期污泥驯化阶段发现,NH_4~+-N浓度高于500 mg·L~(-1)时,污染物去除率明显降低,经HN-AD菌剂生物强化后,耐受NH_4~+-N浓度可高于600 mg·L~(-1)且能保持污染物的高效去除.采用HN-AD生物强化的BCO工艺处理真实猪场沼液,对NH4+-N、TN和COD的平均去除率分别为86. 9%、70. 5%和74. 4%,分别高于传统处理工艺的57. 6%、50. 3%和50. 0%,出水浓度均低于相关污染物排放标准.采用高通量测序技术研究了功能菌富集过程中微生物群落结构的变化规律,结果表明,生物膜内属于HN-AD菌的优势菌由Alcaligenes这一种菌属增加为生物强化后的Diaphorobacter、Acinetobacter和Thauera等多种菌属,且Acinetobacter菌属的相对丰度明显高于接种菌剂.扫描电子显微镜结果也进一步证实了生物强化的存在,紧密附着在填料上的生物膜层表面富集了以杆状和球状为主的HN-AD功能菌.     

碳素纤维去除富营养化水体氮磷的动力学研究

以碳素纤维(CF)为生物膜载体,研究了CF微生物附着特征以及CF和生物碳素纤维(BCF)去除污水氮磷的动力学特征,并考察了曝气量、pH和水力停留时间(HRT)对污水氮磷去除效果的影响。结果表明:CF具有良好的微生物附着特性,挂膜启动快,附着生物量大且生物膜较薄,更有利于生物膜的传质。CF和BCF对氮磷均有较好的去除效果,其中BCF对TP的去除率大于90%,且BCF氮磷去除稳定时间比CF快。CF和BCF氮磷去除规律比较符合指数型,磷的指数拟合度(R2)要优于氮。增加曝气量有利于氮磷的去除,但超过0.15 L/min时TN、NH+4-N的去除率反而降低。弱碱性条件有利于氮磷的去除,最佳HRT为6h。     

植物与微生物协同净化黑臭水体的脱氮性能

该研究通过在室内模拟黑臭水体,建立苦草和梭鱼草独立与混合生态浮床,研究微纳米曝气条件下植物和微生物协同净化黑臭水体的脱氮性能和微生物群落结构特征,并探讨黑臭水体脱氮性能与微生物群落之间的关系。结果表明,苦草组、梭鱼草组和混合组对氨氮的平均去除率为97.83%、98.18%、98.33%,对总氮的去除率为62.38%、72.24%、62.27%,三者都高于对照组。植物对氨氮和总氮的去除不明显,水体微生物和生物膜微生物的硝化和反硝化作用是脱氮的主要途径。第12天、第31天水体和定植棉生物膜的微生物群落的优势门为变形菌门、拟杆菌门、放线菌门和Dependentiae菌门;优势的黄杆菌属、气单胞菌属、假单胞菌属、鞘氨醇单胞菌属、红杆菌属等都属于异养硝化-好氧反硝化细菌。定植棉生物膜的物种数、Chao1指数、Shannon指数都明显高于第12天和第31天的水体微生物群落,水体微生物和种植棉生物膜的优势菌属和群落结构存在一定的差异。实验后期,化学需氧量浓度是影响水体脱氮效果的重要因素。     

A2O-IFAS工艺生物膜微生物群落的演替及分布特性

在A²O-IFAS工艺中探究了不同功能分区生物膜微生物群落的演替及分布特性.利用该工艺处理实际生活污水,总无机氮去除负荷可达（0.10±0.03）kg·m^-3·d^-1,总无机氮和COD的平均去除率分别为70%和79%.高通量测序结果显示,厌氧氨氧化菌Candidatus＿Brocadia在好氧区生物膜中的相对丰度最高（0.43%）,其次是缺氧区生物膜、活性污泥和厌氧区生物膜.此外,絮体污泥与好氧区生物膜有着相似的微生物群落,该结果表明,功能区非生物因素可驱使生物膜微生物群落在缺氧区和厌氧区发生演替现象,DO、C/N比等环境因素和微生物的互作机制是驱动微生物群落分布和厌氧氨氧化菌富集的关键.本研究可为主流厌氧氨氧化菌的富集和技术应用提供重要参考.     

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.     

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.     

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.     

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.