厌氧氨氧化菌的培养与推流式反应器氨厌氧工艺

采用推流式固定化絮体生物反应器培养出高活性的厌氧氨氧化(ANAMMOX)红色颗粒污泥.在稳定运行后,反应器的NH₄⁺-N和NO₂^--N去除率皆大于98%,TN平均去除率为86%,NO₃^--N的生成率约为14%,TN去除负荷达2.56kg/(m³·d).同时考察了进水基质比例对反应器性能的影响,并用扫描电镜观察了颗粒结构.     

有机碳源条件下厌氧氨氧化ASBR反应器中的主要反应

采用5个已稳定运行在厌氧氨氧化状态的ASBR反应器,通过COD、氨氮、亚硝酸盐氮、硝酸盐氮、pH等指标的监测和好氧硝化菌、异养反硝化菌的测定,研究了不同有机碳源条件下反应器中发生的主要反应.结果表明,反应器中存在好氧硝化菌、异养反硝化菌和厌氧氨氧化菌.在COD、氨氮、亚硝酸盐氮等存在条件下,可发生好氧硝化、厌氧氨氧化和异养反硝化反应,先是好氧硝化反应、厌氧氨氧化反应和异养反硝化反应共存,其后依次是异养反硝化反应和厌氧氨氧化反应占主导地位.当C/NO₂^--N在1.7～1.9范围内时,C/NH₄⁺-N为1.7的1号反应器具有最佳的厌氧氨氧化效果,反应结束时其COD去除率、NH₄⁺-N去除率、NO₂^--N去除率分别为100%、81.7%和74.4%.     

高氨氮渗滤液处理的ANAMMOX A2/O工艺研究

通过好氧出水回流到厌氧流化床可以实现厌氧氨氧化过程.对于高浓度氨氮渗滤液,ANAMMOX反应可使ANAMMOX A²/O工艺比普通A²/O工艺的TN去除率提高15%～20%,达32%以上;好氧出水NO-2-N浓度有较大幅度地降低,改善了出水水质.ANAMMOX反应总反应级数为3级,对NH₄⁺-N、NO₃^--N和NO₂^--N的反应级数均为1级,反应速率常数为-3.43E-5 L²·(mmol²·h)^-1.     

部分亚硝化-厌氧氨氧化协同反硝化处理生活污水脱氮除碳

采用SBR-ASBR组合工艺处理实际生活污水,SBR中考察缺氧/好氧时间比及温度对部分亚硝化（partial nitritation,PN）的作用,ASBR中研究COD/NO₂^--N（C/N）对厌氧氨氧化（anaerobic ammonium oxidation,ANAMMOX）协同反硝化脱氮除碳的影响.①控制温度为25℃,在缺氧/好氧时间比为30 min：30 min,单周期交替3次时,NO₂^--N积累率（NiAR）于第22 d为98.06%,比亚硝态氮产生速率（SNiPR,以N/VSS计）为0.28g·（g·d）^-1,同步硝化反硝化去除的TN和COD分别为12.29 mg·L^-1和110.36mg·L^-1.②在缺氧/好氧时间比为30 min：30 min下,温度为15℃时,丝状菌大量繁殖,污泥活性和沉降性变差;温度为30℃时,NH₄⁺-N转化为NO₂^--N比例为86.83%,造成出水NH₄⁺-N浓度过低,不能为厌氧氨氧化提供合适基质浓度;温度为25℃时,出水NH₄⁺-N和NO₂^--N浓度分别为31.58 mg·L^-1和35.04mg·L^-1,匹配厌氧氨氧化基质比.③组合工艺脱氮性能良好,出水TN、NH₄⁺-N和COD浓度分别稳定在13.13、4.83和69.96mg·L^-1,去除率分别为83.10%、93.64%和75.11%.调节ASBR进水C/N为2.5、2.0和1.5时,C/N为2.0时厌氧氨氧化协同反硝化脱氮除碳性能最佳,出水NH₄⁺-N、NO₂^--N、NO₃^--N和COD分别为0.09、0.25、1.04和32.73mg·L^-1.     

厌氧膜生物反应器耦联部分亚硝化/厌氧氨氧化用于污水处理研究

本研究构建了厌氧膜生物反应器（AnMBR）-部分亚硝化/厌氧氨氧化（PN/Anammox）污水处理工艺,以探究AnMBR-PN/A工艺处理效果最佳的水力停留时间（HRT）.AnMBR将厌氧生物处理与膜分离技术相结合实现有机物去除,AnMBR出水NH₄⁺-N通过PN部分转化为NO₂^--N,最终通过NO₂^--N氧化剩余NH₄⁺-N去除.实验结果表明：在HRT=11.2 h时,AnMBR-PN/A工艺化学需氧量（COD）去除率稳定在97%以上,COD转化为CH₄效率超过77.5%,总氮（TN）去除率为78%,出水COD和TN浓度分别低于14和11 mg·L^-1.AnMBR段COD去除率达到95%,平均甲烷产率为0.39 L·L^-1·d^-1.PN段实现了NO₂^--N的高效积累,其出水中NO₂^-/NH₄⁺为0.91±0.11.Anammox段出水中的NO₂^--N、NH₄⁺-N和NO₃^--N浓度分别低于1.0、4.9和5.1 mg·L^-1.高通量测序结果表明PN段氨氧化菌主要为Nitrosomonas,丰度为7.09%,Anammox段主要微生物为Candidatus Brocadia,丰度高达21.01%.本研究构建的AnMBR-PN/A工艺实现了污水处理过程的高效能源回收和深度自养脱氮,研究成果为工程应用提供了理论支撑.     

长期饥饿下厌氧氨氧化反应器的启动与恢复性能

厌氧氨氧化菌生长缓慢,其富集过程困难,是限制厌氧氨氧化技术应用的瓶颈. 因基质供应不足导致的功能菌长期饥饿使厌氧氨氧化（ANAMMOX）反应器稳定运行面临更大挑战. 从长期饥饿的视角出发,重新审视厌氧氨氧化反应器的启动和恢复性能、揭示其抗性机制具有现实意义. 分别采用硝化和反硝化污泥在不同运行条件下成功启动了厌氧氨氧化反应器,分析了长期饥饿后不同启动策略下反应器的恢复性能,及污泥对长期饥饿的耐受机制. 结果表明,接种反硝化污泥的反应器采用高基质浓度低流速运行更稳定,运行85 d后NH₄⁺-N、NO₂^--N和总氮去除率分别高达98.7%、99.3%和89.3%;但长期饥饿（144 d）后采用低基质浓度高流速运行的氮去除性能更优,NH₄⁺-N、NO₂^--N和总氮去除率可在30 d后分别恢复至99.8%、99.8%和93.6%. 研究发现,饥饿期胞外多聚物将厌氧氨氧化菌包裹,有助于保持细胞完整以抵抗长期饥饿胁迫;nirS、hzsA和hdh基因的表达,保证了亚硝酸盐/一氧化氮氧化还原酶、联氨合成酶和联氨脱氢酶的合成,有助于维持厌氧氨氧化活性. 饥饿恢复前后厌氧氨氧化菌属的相对丰度无显著差异,其中Candidatus Kuenenia经30 d恢复后相对丰度提升86%以上,表明其对长期饥饿更具耐受性.     

常温城市污水同步亚硝化-厌氧氨氧化研究

在常温14.7～24.7℃条件下,以城市生活污水为研究对象,采用SBR反应器,通过调整曝气量控制DO浓度为0.05～0.30 mg/Ｌ,进行了同步亚硝化-厌氧氨氧化试验.结果表明,SBR活性污泥反应器可以在常温条件下实现城市污水氨氮的同步亚硝化-厌氧氨氧化反应;DO可以作为其反应终点的指示参数,本试验确定为1 mg/Ｌ;在SBR探索试验中,NH⁺₄-N消耗速率为0.164～0.218　kg/（m³·d）,NO^-₃-N产生速率为0.026～0.036　kg/（m³·d）,TN脱除速率为0.124～0.194　kg/（m³·d）,去除效率为65%～75％;在SBR改进试验中,分别通过提高温度、增设非曝气运行时段和增加厌氧氨氧化菌生物量3个途径,避免了亚硝酸盐的积累,TN去除效率提高至77%～88%.考虑到脱氮速率和实际的工程应用条件,认为增加厌氧氨氧化菌的生物量是提高SBR反应器脱氮效能的优选途径.     

亚硝酸盐对聚磷菌吸磷效果的影响

以厌氧/好氧生化反应器中的聚磷菌为实验对象,探讨了亚硝酸盐对聚磷菌吸磷效果的影响.结果表明:低浓度NO₂^--N可以作为聚磷菌的电子受体,实现NO₂^--N型反硝化除磷,但吸磷总量和吸磷速率明显低于NO₃^--N型反硝化除磷的效果;当NO₂^--N和NO₃^--N共存于缺氧环境时,NO₂^--N对NO₃^--N型反硝化除磷的除磷总量和速率没有影响,但会降低NO₃^--N的消耗量;NO₂^--N型反硝化除磷污泥的好氧吸磷量和速率均低于传统A/O厌氧放磷污泥的效果,但由于它经历了缺氧吸磷和好氧吸磷2个阶段,因此,从吸磷总量或出水水质看,二者相差不大.     

厌氧氨氧化微生物颗粒化及其脱氮性能的研究

利用厌氧颗粒污泥作为种泥,启动SBR反应器,旨在培养厌氧氨氧化颗粒污泥以及研究其脱氮性能.结果表明,水力停留时间(HRT)是富集厌氧氨氧化微生物的1个重要控制因素,以HRT为30 d,第58 d时,SBR反应器就出现厌氧氨氧化现象,与此同时,颗粒污泥由灰黑色变为棕褐色,粒径减小.到第90 d时,成功培养出厌氧氨氧化颗粒污泥,NH⁺₄-N和NO^-₂-N同时被去除,最大去除速率分别达到14.6 g/(m³·d)和6.67 g/(m³·d).从第110　d开始,逐步降低HRT,以提高基质负荷促进厌氧氨氧化菌生长.到目前t＝156　d,HRT降到5　d,氨氮和亚硝酸氮的去除率分别达到60.6%和62.5%,亚硝酸氮/氨氮的比率为1.12.污泥也由棕褐色变为红棕色,形成红棕色的具有高厌氧氨氧化活性颗粒污泥,总氮负荷达到34.3 g/(m³·d).     

补充碳源提取液对人工湿地脱氮作用的影响

为了提高人工湿地的脱氮效率,在不同条件下分别对美人蕉、香蒲及稻杆进行稀硫酸水解,以获得相应碳源提取液.正交实验表明,稀硫酸浓度的提高和水解时间的增加都会导致碳源释碳能力的提高,稻杆在5%稀硫酸溶液中水解30 min以上,释碳能力最高.通过观察,前2 d是系统脱氮反应高峰时段.对以NH₄⁺-N和NO₃^--N为氮源的脱氮过程,随C/N比升高,NO₃^--N和TN去除率增长明显;而NH₄⁺-N受溶解氧制约,去除有限;随C/N比升高,碳源对系统溶解氧的竞争会进一步抑制硝化反应的彻底进行.而对以NO₃^--N为氮源的反硝化过程,补充碳源对TN和NO₃^--N的去除有明显作用;TN去除率由54%提高到95%,NO₃^--N去除率由48%提高到96%;中间产物NO₂^--N的积累与NO₃^--N去除率有关;当NO₃^--N去除率较高时,NO₂^--N无积累.此外,基质反硝化强度也随C/N比升高呈上升趋势,湿地填料细沙层的反硝化强度略高于碎石层.     

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.