微生物降解油田含油污泥中烃类污染物的研究

微生物法治理油污土壤具有成本低、效果好的特点。其原理是微生物利用油烃作为碳源合成自身物质．进行生长繁殖，从而使油烃含量减少。在好氧和厌氧条件下，从油污土壤中分离纯化出4株能降解石油烃类的微生物CH1、CH2、CH3和CH4。经鉴定菌株CH3为假单胞菌属，通过生理特性实验，确定了其生长的最适pH值为7.5，油泥中石油烃类的降解率达85％。按照文中微生物处理舍油污泥的现场施工方案，可使处理后舍油污泥中烃类含量〈1500mg／kg（符合国标GB4284—1984要求）。     

分离海洋不动杆菌及其对石油烃降解性能研究

石油降解菌在石油污染生物修复技术中起到非常重要的作用。本研究分别以渤海湾油污区采集的水样,油样,水油泥混合样为材料富集分离石油降解菌,对其进行生理生化及分子生物学鉴定,并采用GC-MS测定烷烃、环烃、芳香烃等石油烃组分的变化。其中3株菌具有较高石油烃降解能力,16SrRNA序列分析表明该3株菌均与不动杆菌属（Acinetobacter）有99%序列相似性,可初步鉴定为不动杆菌属（Acinetobacter）。3株菌的石油烃降解能力依次为Tust-DM21＞Tust-DC12＞Tust-DW04,对原油成分的降解效果依次为烷烃＞芳香烃＞环烃。其中菌株Tust-DM21为一株高效石油烃降解菌,28℃于富集培养基培养10 d后,对烷烃（C10~C30）的降解率可达98%,对芳香烃和环烃的降解率达88%。研究表明,Tust-DM21菌株对烷烃,环烃,芳香烃都有较强的降解能力,是一株具有较好开发前景的石油降解菌。     

老化油泥处理过程中的微生物群落变化特征

通过在油田现场构建2个中试规模的生物堆(秸秆添加堆和对照堆)对老化油泥进行处理,运用BIOLOGTM系统对油泥中微生物群落水平的生理特性进行分析,测定其微生物代谢活性、代谢多样性及代谢特征差异性,以研究堆体各层次间微生物群落代谢特征的变化规律. 结果表明,与对照堆相比,秸秆添加堆的多项生理特性指标均得到显著提升,其中微生物代谢多样性指数变化范围由1.64～3.02升至2.83～3.29,并且表现出时间上的稳定性和空间层次间的均质性;在微生物碳源代谢特征的差异性分析中,对照堆各层次随时间推移表现出向其外层顺次变化的规律;油泥中总异养菌及石油烃降解菌的数量与环境温度关联性较强,石油烃降解菌的数量在试验末期取得峰值,达4.15×107 CFU/g.      

石油烃降解混合菌的筛选及其降解条件研究

对采集克拉玛依地区的部分石油污染样品进行了富集分离,得到了5组石油烃高效降解混合菌,其中混合菌KL9-1对温度的耐受范围最宽,并且石油烃的降解效率最高。该混合菌在45℃的条件下,通过7 d的降解,稀油的降解率达到43.27%,稠油的降解率达到20.09%。利用单因素试验考察环境因素对混合菌KL9-1降解石油烃的影响,结果表明混合菌KL9-1的接种量、石油烃仞始浓度、初始pH、摇床转速、表面活性剂的添加都会影响石油烃的降解效果,在35℃的条件下,当接种量6.0%、石油烃初始浓度1.5%、仞始pH 7.5、摇床转速120 r/min及添加200 mg/kg Tween80表面活性剂时,稀油和稠油的降解率都达到最高,其中稀油的降解率可以达到62.49%,稠油的降解率达到40.36%。     

石油降解菌的筛选及其降解效率的研究

从石油污染土壤中分离纯化得到1株特征明显的石油降解菌,研究了不同的油含量、温度及添加物等因素对石油降解菌的脂酶活性和除油率的影响。结果表明:实验菌株属于假单胞菌属;脂酶活性与除油率正相关;除油率随油浓度的升高而降低;石油降解率通常随温度的降低而降低;添加蔗糖不利于石油的降解;添加表面活性剂吐温80有利于脂酶的产生和代谢。     

解脂耶氏酵母菌处理含油废水的研究

利用解脂耶氏酵母菌在最佳降解条件下分别处理色拉油、餐饮油等油脂类废水和柴油、机油等石油类废水50 h.结果表明:该菌株能快速降解油脂,在油质量浓度≤2 000 mg/L时生化反应属于一级反应,对色拉油、餐饮油中油脂的降解率分别为93.30%和85.08%,同时菌体细胞大量生长,其生物量分别为1 652.7和1 940.0 mg/L,废水处理过程中pH稳定在3～4之间;但该菌对石油的降解率低,生成的生物量少,处理后废水pH维持在6～7之间.解脂耶氏酵母菌对油的降解与自身生物量的生成有关,该菌株降解油脂类和石油类物质的途径不同,产生的脂酶系统不同,使得废水的pH变化规律不相同.      

油田含油污泥中烃类污染物的微生物降解

在好氧和厌氧条件下,从油田被污染的士壤中分离纯化出四株能降解石油烃的微生物菌株CH1—4,为假单胞菌属。通过生理特性实验,确定了其生长的最适PH值,油泥中石油烃类的降解率达85%。按照文中微生物处理含油污泥的现场施工方案,可使处理后油泥中烃类物质含量小于3000mg/Kg(符合国标GB4284—84要求)。     

油基泥浆钻井岩屑中高效石油降解菌的筛选及其降解特性

以四川珙县油基泥浆钻井岩屑为研究材料,筛选出10株能以石油为碳源和能源生长的降解菌,并以石油降解率为筛选依据,筛选出2株高效降解菌(C-1、C-2),通过16S r DNA序列分析对其进行初步鉴定和降解特性研究。结果表明:C-1、C-2的降解率为39%~40%,分别为芽孢杆菌属Bacillus sphaericus、Bacillus subtilis;降解特性分析表明:在接种量为10%、pH为7、温度为35℃、n(N)∶n(P)=12∶1、盐浓度为0.5%的最优条件下,C-1和C-2对石油的去除率分别为46.08%、48.94%,补充N、P后降解率基本未变化,说明岩屑中大部分可生化降解成分已全部降解,部分难降解的物质(如长链烷烃、多环芳烃等)仍在岩屑中。     

高效石油降解微生物堆制法处理油污土壤

采用高效石油降解菌复合堆制法来处理油污土壤(土样1和土样2),对过程中的基本参数如温度、pH和水分进行适时的调控;定期分析添加的高效石油降解菌菌数变化以及总油含量变化,结果表明这种堆制方式对油污土壤的修复作用十分明显,扣除其他因素的影响,生物修复的去除率分别为42.9%和44.3%。     

嗜油菌NY3与土著菌FF生物强化降解石油烃的协同机理研究

优良菌与土著菌的相互作用关系是影响生物强化降解石油烃效果的关键因素。前期研究发现，嗜油菌NY3生物膜对2 g/L含油废水中石油烃的降解率可保持在83%～95%,处理阶段与恢复阶段生物膜上NY3菌及FF菌的生物量存在此消彼长的变化趋势，本文旨在探明该变化趋势的产生机理。研究结果表明：NY3菌对长链烷烃(C30～C34)的降解率高于FF菌，FF菌对中长链烷烃(C13～C29)、蒽、菲的降解率高于NY3菌；NY3菌在中性、弱碱性和低盐度环境下对石油烃的降解率高于酸性和高盐度环境，FF菌在中性、弱酸性、高盐度环境下对石油烃的降解率高于碱性和低盐度环境；NY3菌分泌的鼠李糖脂和绿脓菌素可促进FF菌的生长，但FF菌分泌的海藻糖脂可抑制NY3菌的生长。据此可知，NY3菌与FF菌能协同降解石油烃，但因胞外分泌物对NY3菌和FF菌的作用不同，致使NY3菌和FF菌的生物量出现此消彼长的变化趋势。     

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.