Diaphorobacter sp.PDB3菌好氧反硝化脱氮特性

研究了Diaphorobactersp.PDB3菌在发酵罐内好氧反硝化特性.经过分析比较确定柠檬酸钠是好氧反硝化最佳碳源,采用响应面法对影响好氧反硝化的条件（温度、pH值、发酵罐转速、碳氮比）进行优化,构建出好氧反硝化数学模型.本模型预测值与实测值相关性非常好,预测PDB3菌发酵罐内好氧反硝化最佳条件：pH值、碳氮比、温度、转速分别为7.3、8.1、30.1℃、299.9r/min,最佳好氧反硝化比速率为2.25h^-1.验证实验表明,PDB3菌在发酵罐内好氧反硝化比速率为2.12h^-1,误差仅为5.8%.通过氮平衡分析表明,29.0%的总氮去除量转化为胞内氮,氮损失占总氮去除量的67.3%,细胞主要通过好氧反硝化作用和细胞同化作用脱氮.荧光定量PCR分析测定好氧反硝化过程中脱氮基因（napA,nirS,cnorB,nosZ）的表达水平以进一步解释脱氮特征,为Diaphorobacter sp.PDB3菌的实际应用提供一定的理论依据.     

耐冷嗜碱蒙氏假单胞菌H97的鉴定及其好氧反硝化特性

结合形态、磷脂脂肪酸及16S rRNA基因序列分析鉴定了分离自贵州冬水田的好氧反硝化菌株H97,采用模拟废水探讨了不同温度、接菌量、C/N、初始p H和碳源种类对菌株H97反硝化能力的影响.结果表明,菌株H97为蒙氏假单胞菌(Pseudomonas monteilii),目前国内外尚无蒙氏假单胞菌具有耐冷嗜碱好氧反硝化作用的研究报道,是对好氧反硝化菌的补充;该菌的最适脱氮条件为15℃、p H 9.0、C/N 15、接种量1.5×106CFU·(100 m L)-1,碳源丁二酸钠.此外,菌株H97在p H为7.0~11.0时,均表现出良好的脱氮能力,对硝酸盐氮和总氮的去除率分别可达91.21%和79.10%以上,初始p H为12.0时,对硝酸盐氮和总氮的去除率仍分别达64.75%和36.78%,表现出了较强耐碱能力.在初始硝酸盐氮浓度为50.0 mg·L-1和最适脱氮条件下,菌株H97在48 h内对硝酸盐氮和总氮的去除率可达97.69%和96.32%.同时,H97对温度适应范围广,在15~40℃均具有较强好氧反硝化能力,是1株耐冷嗜碱好氧反硝化细菌,在碱性氮污染水体处理中具有较好的应用潜力.     

富氧条件流离反应器内好氧反硝化特征

基于连续流A/O流离生物膜反应器内同步硝化反硝化的研究结果,以流离生物膜内菌群为整体研究对象,在富氧条件下,对依赖不同氮源生存的细菌的活性、以及具有的反硝化特征进行了研究.研究结果表明,温度25~30℃、溶解氧4.0~6.0 mg·L-1条件下,低碳氮比废水在硝化菌和好氧反硝化菌共同作用下,总氮和氨氮浓度稳步下降,亚硝酸盐和硝酸盐在试验持续时间内无明显积累现象.再分别以硝酸盐氮和亚硝酸盐氮为氮源,在高温(40℃)条件下18 h内即被完全去除,证明流离生物膜内的好氧反硝化菌脱氮效果好,且对高温水环境耐受力强.相对于单一菌群的反硝化研究,以多种细菌整体为研究对象的试验研究具备实际应用的可能性.     

溶解氧对Biolak型A2O工艺脱氮除磷性能的影响

通过对Biolak型A2O工艺处理生活污水工程应用的研究,考察了好氧段溶解氧(DO)浓度对该工艺脱氮除磷的影响.试验结果表明,DO浓度变化对系统COD、NH+4-N处理效果的影响不大,而对系统总氮及总磷的去除效果影响显著.当DO浓度控制在0.80~1.50 mg·L-1之间时,系统总氮去除效果最佳,可以达到69.5%,系统好氧段可实现同步硝化反硝化除氮.通过对系统氮进行物料衡算发现,23.7%的总氮通过好氧段多级A/O反硝化脱氮去除.当DO浓度为1.00~3.00 mg·L-1时,总磷(TP)去除率较高,可以达到74.0%.DO浓度控制在1.00~1.50 mg·L-1之间时,系统脱氮除磷效果最佳,此时TN、TP的去除率分别为68.9%、73.7%,二级生化处理段出水TN、TP分别为12.02、0.95 mg·L-1.     

缺氧反硝化-好氧间歇试验处理印染PVA退浆废水

系统研究了投加硝态氮NO3^--N的反硝化工艺在缺氧，好氧，生物碳间歇试验中处理印染PVA退浆废水的效果。结果表明．按照缺氧反硝化投加硝态氮NO3^--N比缺氧水解-生物膜法对CODcr的去除率：缺氧池，好氧池均提高了30％．缺氧反硝化工艺二沉池出水经过生物碳处理后CODcr的去除率达到90％。缺氧反硝化，好氧的降解速率常数分别由缺氧水解，好氧的0．0126h^-1，0．0345h^-1，提高到0．080h^-1,0．112h^-1,NO3^--N与CODcr之比为0．20是处理印染PVA退浆废水成功的关键。     

移动床生物膜系统SND影响因素研究

采用移动床生物膜反应器实现了稳定同步硝化反硝化脱氮.实验研究了C/N对同步硝化反硝化脱氮的影响,结果表明,随着C/N的增加,同步硝化反硝化脱氮效率提高,在好氧条件下总氮去除率最高达到92.9%,但当C/N=12时,TN去除率提高并不明显;实验研究了pH对氨氮和TN去除效果的影响,结果表明,氨氮去除pH适宜区域为8.03~9.01,TN去除pH适宜区域为8.03~8.55;实验分析了实际生活污水中碱度和pH值对脱氮效果的影响,并对比研究了理论碱度和实际碱的关系,结果表明碱度和pH在同步硝化反硝化脱氮中对氮素去除的影响不大,不需要额外调节系统pH,也不需要在反硝化过程中补充碱度.单个周期内同步硝化反硝化过程中pH值和碱度变化规律的研究表明,pH可以指示SND中氮素转化过程.     

回流液反硝化状态转变及脱氮效果提升研究

为探究工艺特性对硝化混合液液相主体及其中污泥分别从有氧、好氧呼吸状态转变为无氧、反硝化状态的影响。该文在A/O系统中研究了不同DO条件下,内回流液从好氧硝化状态转变为缺氧反硝化状态过程中液相主体物质及反硝化速率的变化,以及在不改变HRT及其他条件的情况下,在A/O系统上分别增设前置预反硝化池、后置预沉淀池,且好氧池均改用渐减曝气法,探究对系统脱氮效果的影响及氮的去除途径。结果表明：在保证充分硝化下,相对好氧池溶解氧较低时,无论回流液液相主体还是污泥的状态转变都较快,污泥反硝化速率在短时间内迅速提升;增设前置预反硝化池或后置预沉淀池可以将系统的脱氮效率从79%分别提高至83%和89%,且后置预沉淀池改善了沉淀池的污泥沉降性能。好氧池采用渐减曝气,系统脱氮率也会有所提高,无论是后置预沉淀池还是前置预反硝化池,都使好氧混合液中污泥快速转变为反硝化状态,提高了缺氧池污泥反硝化速率以及系统脱氮效率。     

异养硝化-好氧反硝化菌的筛选及脱氮性能的实验研究

通过极限稀释和显色培养基相结合筛选的方法,从畜禽养殖废水样品中筛选到1株同时具有异养硝化-好氧反硝化双重功能的细菌CPZ24.该菌株革兰氏染色呈阳性,杆状,菌落颜色为橙红色.经形态、生理生化特性,16S rDNA序列分析,初步鉴定该菌为嗜吡啶红球菌(Rhodococuus pyridinivorans).对该菌进行异养硝化功能和好氧反硝化功能进行研究,结果表明,在异养硝化过程中,该菌可将培养基中的氨氮全部去除,其中对总氮的去除率可达98.70%;在好氧反硝化过程中,该菌对硝酸盐氮的去除率可达到66.74%,总氮的去除率达到64.27%.此高效脱氮降解菌可实现自身同步硝化-反硝化脱氮功能,能够独立完成生物脱氮的全过程.     

2株好氧反硝化菌株的分离鉴定与脱氮特性研究

好氧反硝化细菌是微生物脱氮技术的重要支撑,具有处理周期短、脱氮效率高等优势。该文从SBR系统中分离出2株好氧反硝化细菌,经过生理生化测定、16S rDNA鉴定以及同源性对比,确定SBR3-3是肺炎克雷伯氏菌,SBR3-4是植生拉乌尔菌,并对其生长特征、16S rDNA鉴定以及不同投加量、不同温度条件下脱氮性能进行了试验研究。试验结果表明,在25～30℃条件下2株菌对总氮去除率最高分别为70%和69%,NO_3~--N平均去除率为95%;在5～10℃条件下对总氮去除率最高分别为41%和43%,NO_3~--N最高去除率分别为57%和55%。在运行过程中,受C/N比和温度的影响,NO_2~--N有一定程度的积累。当SBR3-3和SBR3-4菌液的添加量为2 mL∶2mL时,对NO_3~--N和总氮的去除效果最好,总氮的去除率达到75%。相关研究成果对比来看,所筛选的2株菌在25～30℃条件下脱氮效果较好,具有良好的好氧反硝化效能。     

两株高效好氧反硝化细菌的分离鉴定及其脱氮效率

从水稻土和活性污泥中分离到两株可以在好氧条件下进行反硝化作用的细菌ZW23和ZW27.通过对这两株细菌的形态观察和生理生化特征,以及16S rDNA序列测定认为菌株ZW23和ZW27分别是假单胞菌属类产碱杆菌(Pseudomonas pseudoalcaligenes)和假单胞菌属门多萨菌(Pseudomonas mendocina).好氧培养条件下,在初始氮源约为280.00mg·L-1的反应体系中,两株细菌在12h内均引起体系中总氮显著下降,削减率分别达66.43%和65.54%,其余总氮几乎全部转化为内源氮.脱氮速率分别达到约21.72mg·L-1·h-1和22.31 mg·L-1·h-1,比现已分离出的兼性好氧反硝化细菌的脱氮速率要快得多.反应过程中没有检测到亚硝态氮和氨态氮的积累.菌株ZW23和ZW27是两株典型的高效兼性好氧反硝化细菌,具有重要的应用价值.     

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.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

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

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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.     

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.