1株α-蒎烯降解菌的分离鉴定及降解特性研究

从处理废气的生物滤塔内筛选到1株能高效降解α-蒎烯的菌株PT.通过菌落形态、生理生化特征、16SrRNA基因序列相似性分析及Biolog鉴定等方法,确定该菌株属于荧光假单胞菌(Pseudomonas fluorescens).菌株PT最佳生长条件为: NaCl浓度0.00%、pH7.13、温度25.5℃,降解速率达到最大值5.22mg/(L×h).降解过程符合Haldane’s抑制生长动力学模型,最大比降解速率为0.0364h-1.菌株PT能不同程度地降解一些分子结构较为简单或与α-蒎烯结构相似的工业有机污染物.代谢产物分析表明,菌株PT在降解α-蒎烯的过程中产生柠檬油精、紫苏酸等结构较为简单的物质,它们最终被完全矿化为CO2或合成细胞自身组成物质.碳平衡分析表明,底物有机碳含量完全矿化和转化为细胞生物量的比例分别为64.83%和30.37%.     

1株氯苯高效降解菌的分离鉴定及降解特性

分离筛选到1株能以氯苯为唯一碳源和能源的菌株LW26,根据菌株的形态、生理生化特征、16S rRNA序列分析以及Biolog鉴定,确定该菌株为戴尔福特菌(Delftia tsuruhatensis),其为新发现的具有氯苯降解能力的菌株,并且该菌株能降解BETX、正己烷和环己烷等常见的有机污染物.实验考察了温度、pH、氯苯初始浓度、Cl-浓度等因素对菌株生长和降解性能的影响.结果表明,该菌株较为适宜的生长和降解条件为:温度25℃、pH 7.0;底物耐受浓度高达500 mg·L~(-1);当Cl-浓度超过0.14 mol·L~(-1)时,菌株生长会受到抑制.利用Haldane模型对实验数据拟合得到菌株LW26的最大比生长速率μmax和最大比降解速率γmax分别为0.42 h~(-1)和2.53 h~(-1).利用GC-MS进行中间产物分析,结果表明菌株LW26降解氯苯的过程中产生邻氯苯酚,结合邻苯二酚双加氧酶活性分析,推测氯苯经历邻位开环、脱氯、氧化等过程,最终矿化为CO2或转化为生物质.     

1株BTEX降解新菌株的分离鉴定及其降解特性研究

取自炼油污水处理厂曝气池的活性污泥经过苯系物定向驯化后,选育到1株能同时高效降解苯、甲苯、乙苯和邻二甲苯(BTEX)的菌株byf-4,基于形态特征、生理生化、16S rDNA序列系统学分析和Biolog鉴定,可确定该菌株为染料分枝杆菌Mycobacterium cosmeticum,其为新发现的1株具有降解BTEX性能的菌株.该菌株最佳生长温度和pH分别为30℃和7.0,其对4种苯系物的降解优先顺序为苯、甲苯、乙苯、邻二甲苯;菌株降解苯、甲苯、乙苯和邻二甲苯的比耗氧速率分别为165.3、170.5、49.3和57.4 mg.(min.mg)-1;菌株降解BTEX的过程遵循Haldane动力学模型,对苯、甲苯、乙苯和邻二甲苯的最大比降解速率分别为0.518、0.491、0.443和0.422 h-1,菌株最大比生长速率分别为0.352、0.278、0.172和0.136 h-1.     

高效降酚菌株Ochrobactrum sp. CH10生长动力学和苯酚降解特性的研究

菌株Ochrobactrum sp. CH10是从北京元大都城垣遗址处的人工湿地筛选到的高效苯酚降解菌.以苯酚为唯一碳源和能源对其进行了生长和苯酚降解特性的研究.该菌生长和降解苯酚的适宜条件为30℃、初始pH 7.0、接种量为5%.在该条件下,初始苯酚浓度为400 mg·L-1,24 h时苯酚完全被降解;初始苯酚浓度为900 mg·L-1时,44 h的降解率为92.3%;初始苯酚浓度为1 000 mg·L-1,48 h时的降解率为82.2%.对该菌株苯酚降解动力学过程进行模拟,符合基质抑制型的Haldane模型,各参数分别为:υmax(最大比降解速率)0.126 h-1,KS(半饱和常数)23.53 mg·L-1,KI(抑制常数)806.1 mg·L-1.该菌在苯酚中的生长动力学符合Andrews模型,表现出与苯酚降解相似的趋势.该菌为目前所发现的Ochrobactrum菌属中苯酚降解能力最强的菌株.该菌株在高效处理含酚废水方面具有广阔的应用前景.     

两株甲苯高效降解菌降解性能的比较

利用自行研制的玻璃龙头塞三角瓶对2株甲苯降解高效菌的甲苯降解性能进行了测定和比较.在纯培养和以甲苯为单一碳源条件下进行摇瓶实验,结果发现,在初始液相浓度为0～75mg/L范围内,甲苯不会对2株菌的甲苯降解活性产生抑制作用.2株菌对甲苯进行降解的适宜pH值约为7～8,适宜温度范围为28～37℃.比较适宜条件下甲苯降解动力学参数可知,2号菌的甲苯降解性能优于1号菌,其最大比基质降解速率和表观最大比增殖速率分别为0.40h-1和0.18h-1.     

两株表面活性剂降解菌株的分离、鉴定及降解特性

从洗涤剂厂排污口附近的河水中,分离纯化到2 株能分别以壬基苯基聚氧乙烯醚(Triton X-100)和十二烷基聚氧乙烯(23)醚(Brij-35)为唯一碳源和能源生长的降解菌株,分别命名为菌株T-1 和B-2.根据菌落特征、菌体形态、生理生化反应分析以及16S rDNA 序列分析,确认菌株T-1 为苍白杆菌属(Ochrobactrum sp.),菌株B-2 为伯克霍尔德菌(Burkholderia).在底物浓度较低(50~100 mg/L)时,Triton X-100降解速度＜Brij-35;在底物浓度较高(200~1500 mg/L)时,Triton X-100 的降解的速度＞Brij-35.T-1 在pH6~8,B-2 在pH7 时生长最好;最适生长温度分别为25,30℃.2 株菌对氮源利用广泛.Zn²⁺、Ca²⁺、Al³⁺、Fe³⁺对T-1 和B-2 均有轻微的抑制作用,Cu²⁺均有毒性.     

某污水处理厂中17α-乙炔基雌二醇降解菌的分离鉴定及其降解特性

由北京某污水处理厂活性污泥中分离出以17α-乙炔基雌二醇(EE2)为唯一碳源生长的单一菌株,经16SrRNA基因序列分析鉴定为香茅醇假单胞菌(Pseudomonas citronellolis),命名为SS-2菌株.研究表明,SS-2菌株在7d内对初始浓度为4mg·L-1EE2的降解率为93.6%,对初始浓度为2mg·L-1雌酮(E1)、或17β-雌二醇(E2)的降解率为99%,对雌三醇(E3)没有降解能力.SS-2菌株降解雌激素的反应符合动力学一级反应定律,其降解速率常数分别为0.236h-1(E1)、0.221h-1(E2)和0.013h-1(EE2).SS-2菌株降解E2的过程中,检测到E1的生成,并且生成的E1亦可被SS-2菌株降解;该降解过程中类雌激素活性随着E2与E1浓度的降低而显著减少,表明SS-2菌株不仅可以降解E2与E1,而且可以降低降解过程中的类雌激素活性;反应进行240h后,仍表现出少量类雌激素活性,推测是因残留的E2或E1所致,表明尚且未知的中间产物或生成物的类雌激素活性远低于E2与E1.     

苯酚降解菌的分离及降解特性研究

从扬子乙烯集团废水处理系统曝气池中的活性污泥驯化分离得到一株能快速降解苯酚的菌株,初步鉴定其为假单胞菌属菌株。该菌株在5℃-35℃范围内时都可以有效降解并矿化200mg／L的苯酚,最适宜的生长温度为25℃左右;菌株在pH为5～9范围内可以降解200mg／L的苯酚,偏碱性的条件下比酸性条件更适合细菌生长;培养过程中振荡速率大于120r／m时降解速率最大。当苯酚的初始浓度超过1000mg／L时,降解菌的生长受到抑制,不能有效降解苯酚。     

潘多拉菌LX-1菌株对二氯甲烷的降解特性研究

从某污水处理厂的活性污泥中分离筛选到一株能以二氯甲烷为唯一碳源和能源生长的好氧降解菌LX-1(Pandoraea pnomenusa LX-1).单因素试验表明,适宜菌株LX-1生长的pH和环境温度分别为7.0和35℃,培养基里添加1%的NaCl有助于LX-1的生长及对DCM的降解,二氯甲烷的最大耐受浓度达到了1500mg·L-1.菌株LX-1降解1mg二氯甲烷能产生0.8083mg Cl-和0.3838mg CO2,脱氯率和矿化率分别达到了96.8%和74.15%.培养液中可溶性有机碳含量随降解时间变化呈上升趋势,pH呈下降趋势,结合相关文献报道,推测菌株在代谢二氯甲烷的过程中产生了水溶性小分子酸类(如甲酸)等结构较为简单的有机物,它们最终将被矿化为CO2、H2O和细胞生物量.     

喹啉降解菌Rhodococcus sp.QL2的分离鉴定及降解特性

从某焦化厂生物处理系统的活性污泥中驯化、分离出1株能以喹啉为唯一碳、氮、能源生长代谢的菌株QL2.经过对其形态特征、生理生化特征和16S rRNA序列分析鉴定该菌株为红球菌属(Rhodococcus sp.).研究表明,菌株QL2利用喹啉生长的适宜温度为35～42℃,培养基初始pH为8～9,摇床转速为150 r/min.外加氮源能促进菌株的生长,其中无机氮比有机氮、铵态氮比硝态氮更利于细菌的生长.在喹啉初始浓度为60～680 mg/L范围内菌株QL2降解喹啉符合零级动力学方程.喹啉初始浓度为150 mg/L时在8 h内完全降解,TOC去除率14 h内可达到70%.降解过程中产生有颜色的物质,且杂环上的氮原子以氨氮的形式被释放.通过HPLC及GC/MS分析出喹啉降解过程中的主要中间产物为2-羟基喹啉.该菌底物利用范围广,能降解苯酚、萘、吡啶等多种芳香族化合物.     

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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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.