烷烃高效降解菌的广谱降解性能研究

针对分离、筛选出的正烷烃高效降解菌株C-14-1的广谱降解性能进行研究.结果表明,C-14-1菌株不仅对较高浓度(大于50mg/L)的正烷烃降解能力显著,对低浓度(约2mg/L)的正烷烃系列同样具有较好的降解效果,并且降解率和烷烃的碳原子数紧密相关.经过48h的发酵,当烷烃碳原子数为10～22时,降解率基本上大于85%;当烷烃碳原子数大于22时,降解率降低到15%～78%.C-14-1菌株对浓度很低(约0.3mg/L)的原油也显示出一定的降解效果.当烷烃碳原子数为12～22时,降解率平均为65%;当烷烃碳原子数大于22时,降解率降低为平均28.3%.当试验系统处于限氧状态时,C-14-1菌株对5α-雄甾烷和六甲苯基本上无降解作用,但对姥鲛烷的降解率却高达97%.在供氧充足的通气状态下以5α-雄甾烷为唯一碳源时,C-14-1菌株对5α-雄甾烷的降解率达到77%.当与其它低碳原子数的正烷烃共存时,5α-雄甾烷更容易被C-14-1菌株通过协同代谢而降解.通气培养48h时,其降解率达到100%;即使在限氧状态下,其降解率也能达到64%.     

三种细菌降解直链烷烃的效果及降解动力学研究

从石油污染土壤中筛选出克雷白氏菌属(Klebsiellasp.,A5)、假单胞菌属(Pseudomonas sp.,A6)和无色杆菌属(Achromobacter sp.,A10)3种石油烃降解菌,研究了其对石油烃中不同碳原子数的3种直链烷烃(正十四烷、正十五烷和正十六烷)的降解效果。结果表明:克雷白氏菌属对3种烷烃的降解能力相对较差;假单胞菌属对3种烷烃的降解效果较好,5d的降解率均达到80%以上,且对正十六烷的降解率高达95.9%;无色杆菌属对正十五烷和正十六烷也有较高的降解率,对正十五烷的降解率达80%以上,对正十六烷的降解率达90%以上。选取降解效果最好的假单胞菌属对正十六烷进行降解动力学研究,结果表明:当正十六烷初始含量为100mg/L、200mg/L、400mg/L和800mg/L时,其降解动力学与一级动力学方程拟合效果良好,其降解半衰期为1.79~3.22d;同时,降解过程中菌体浓度的变化显示,环境中的正十六烷含量越低,菌体的对数生长期越短,当正十六烷含量为100mg/L和200mg/L时,A6菌的对数生长期仅持续1d左右就进入稳定期,而当正十六烷含量达到400mg/L以上时,A6菌的对数生长期持续2d后,生长菌群总数处于平坦阶段。     

丁二腈高效降解菌的筛选及其降解性

以丁二腈为唯一碳源和氮源,从石化腈纶废水及其处理构筑物的生物膜中,分离、筛选出2株高效降解丁二腈的菌株:J-1-3和J-13-1.经形态学观察和生理生化特征研究,两者均被鉴定为假单胞菌(Pseudomonas spp.).通过摇瓶试验得出2菌株的最适生长条件:温度为30℃,摇床转速(间接反映通气量)为250 r/min,接种量为0.1%,初始pH为6.在最适生长条件下,分别对不同初始浓度丁二腈进行降解率试验.结果表明,2菌株对丁二腈的降解能力强,尤以J-13-1更为显著.当丁二腈的初始浓度约为6000mg/L、8000mg/L和10000mg/L时,J-13-1菌株对丁二腈的降解率分别在12.5h、14h和16h时达到100%.     

2,6-二叔丁基酚降解菌的降解特性研究

从腈纶废水处理构筑物的生物膜中分离、筛选得到1株能降解2,6-二叔丁基酚的菌株,经驯化,其对2,6-二叔丁基酚的降解率提高了26%,具有了较高降解能力.经形态和生理生化鉴定,该菌株属于产碱菌属(Alcaligenes sp.).通过摇瓶试验考察了生长条件对菌株的生长和底物降解的影响,得出该菌株的最适生长条件为温度37℃,初始pH为7.0,接种量为0.1%.在该条件下,对初始底物浓度为100mg/L的降解过程进行了考察,结果表明其11d的降解率达62.4%,而且降解过程符合Eckenfelder动力学模型,半衰期为9.38d.还对不同初始底物浓度对菌株降解性能的影响进行了研究,结果表明最佳初始底物浓度为200mg/L,当小于该值时,初始底物浓度的增加促进该菌株的生长和底物的降解,而当大于这个值时,则起抑制作用.     

高效苯酚降解菌Bacillus sp.L5-1的分离及其降解特性

从污水处理厂活性污泥中分离筛选出一株高效苯酚降解菌L5-1,经菌落形态观察和16S rDNA基因测序,结果表明菌株L5-1为蜡样芽胞杆菌（Bacillus cereus）,美国国家生物信息中心（NCBI）的注册号为MN784421.将苯酚设置为唯一碳源,对其生长和苯酚降解特性展开研究.结果表明：菌株L5-1在10%接种量、温度30~35℃、pH值7~8的条件下,均能高效降解培养基中苯酚（培养基体积为100mL,初始苯酚浓度为500mg/L,14h时降解率>93%）.而在最优降解条件下（10%接种量,培养温度为35℃,pH值7.0,NaCl浓度为1%）,初始苯酚浓度为500mg/L,菌株在14h内的苯酚降解率可达97.1%;而当初始苯酚浓度为1000mg/L,菌株也可在46h内达到97.71%的降解率.运用Haldance方程动力学模拟菌株在不同浓度苯酚下的生长过程,其最大比生长速率为0.355h^-1,半饱合常数104.27mg/L,抑制常数为322.83mg/L,R²=0.997.菌株L5-1为目前已报道的Bacillus菌属中降解苯酚能力较强的菌株,为实际处理含酚废水中提供理论参考.     

微囊藻毒素-R R 高效降解菌的分离鉴定及降解特性

从蓝藻爆发期的上海市淀山湖表层水体中筛选分离降解微囊藻毒素-RR(MC-RR)的细菌,研究其降解特性。根据分离菌株的细胞形态结构、生理生化特征及其16S rDNA序列分析鉴定降解菌,高效液相色谱法测定该菌株降解MC-RR的能力。分离菌株DHU-38(GenBank序列登录号为HM047515)属荧光假单胞菌(Pseudomonas fluorescens)。微囊藻毒素降解实验结果表明,该菌株能在以MC-RR为唯一碳源、氮源的无机盐培养基中生长,6 d内可将初始质量浓度为20 mg/L的MC-RR降解为6.23 mg/L,降解效率达到69%。菌株DHU-38的最适生长温度是30℃,最适生长pH值为7.0。酵母粉、蛋白胨、葡萄糖等营养物质可以明显促进菌株对MC-RR的降解效率,尤其是加入100 mg/L酵母粉后,6 d降解率达到89.6%。     

一株氯苯优势降解菌的降解条件优化

以氯苯降解率为降解效果指标,以降解温度、初始pH、降解时间、接种量和氯苯初始浓度为影响因素,对实验室保藏的一株氯苯优势降解菌株Lysinibacillus fusiformis LW13降解氯苯的降解条件进行优化。单因素试验结果表明,该降解菌株对氯苯的适宜降解条件分别为:温度20～40℃,pH为8.0,降解时间4 d,接种量2%～4%,氯苯初始浓度60～140 mg/L。以降解温度、氯苯初始浓度和接种量这三个显著影响因素进行正交试验,结果表明各影响因素的主次顺序为降解温度>氯苯初始浓度>接种量,最佳降解条件为降解温度35℃、氯苯初始浓度100 mg/L和接种量4%,最佳降解条件下氯苯降解率可高达93.8%。     

高效耐盐柴油降解菌的筛选、鉴定及降解基因

在辽河口湿地油田石油污染土壤中分离到一株耐盐柴油高效降解菌株L7,经过形态学观察、16S r RNA基因序列分析鉴定菌株L7属于不动杆菌属(Acinetobacter).该菌株能够以柴油、十五烷、十六烷、十七烷、十八烷、十九烷、二十烷和二十二烷为唯一碳源和能源生长.在柴油无机盐培养基中,菌株的最适生长温度为30°C,最适pH值为7.0,菌株L7在pH 6.0~9.0,盐度范围为3%以内都可以生长.与其他的菌株相比较,菌株L7在柴油中生长更加迅速,在最适培养条件下培养4d后菌株的OD600nm就可以达到4.0,通过紫外分光光度法测定培养5d后,柴油的降解效率为61.5%.通过对菌株L7的全基因组测序和分析,在基因组上找到一个烷烃羟化酶基因alk B,将基因克隆到p ME6032质粒上,并电转至不依赖柴油生长的恶臭假单胞菌KT2440中,含有该质粒的KT2440菌株能够在柴油无机盐培养基中生长.GC-MS检测确定了菌株L7及其alk B基因对饱和烷烃的降解能力.推测菌株L7中烷烃降解途径为末端氧化途径,是由烷烃羟化酶催化反应生成相应的醇,最终通过β-氧化反应实现完全矿化.     

Rhodococcus sp. T3-1菌株降解乙草胺的特性

以乙草胺为唯一碳源,通过摇瓶培养研究了Rhodococcus sp.T3-1对乙草胺的降解特性.结果表明,菌株T3-1降解乙草胺的最适温度为37℃,且其在pH值6~10的范围内对100mg/L乙草胺的降解率均在96%~97%之间.该菌株在接种量为5%条件下,14h内可将200mg/L的乙草胺降解95.5%;乙草胺的降解速率与乙草胺初始浓度呈负相关,与菌株T3-1的初始接种量呈正相关.菌株T3-1还可以降解丁草胺,但不能降解丙草胺、异丙草胺和吡草胺.     

双酚A降解菌的分离鉴定及其降解特性

从好氧堆肥反应器的渗滤液中分离到1株降解内分泌干扰物双酚A的细菌B-16.经过对其形态特征、生理生化以及16S rDNA序列分析,该菌株初步鉴定为Achromobacter xylosoxidans.通过摇瓶实验考察了生长条件对双酚A降解的影响,得出该菌株降解双酚A的最佳条件是接种量为0.6%,初始pH值为7.0,温度为30℃.在该条件下,对菌株B-16对不同初始浓度双酚A的降解反应过程进行动力学分析,该降解反应在低浓度(3、5、10mg/L)时符合一级反应动力学特征,而在较高底物浓度(20 mg/L5、0 mg/L)下不能用一级反应动力学描述,双酚A的降解率在初始浓度为10 mg/L时达到最大值46.93%.     

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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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.     

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.     

Sorption of 1-naphthol by plant cuticular fractions

The contribution of aliphatic-rich plant biopolymer to sorption of hydrophobic organic compounds is significantly important because of their preservation and accumulation in the soil environment,but sorption mechanism is still not fully understood.In this study, sorption of 1-naphthol by plant cuticular fractions was examined to better understand the contributions of respective fraction.Toward this end,cuticular materials were isolated from the fruits of tomato by chemical method.The tomato cuticle sheet consisted of waxes (6.5 wt%),cuticular monomer (69.5 wt%),and polysaccharide (24.0 wt%).Isotherms of l-naphthol to the cuticular fractions were nonlinear (N value (0.82-0.90)) at the whole tested concentration ranges.The KodKow ratios for bulk cuticle (TC1),dewaxed cuticle (TC2),cutin (TC4),and desugared cuticle (TC5) were larger than unity,suggested that tomato bulk cuticle and cutin are much powerful solption medium.Sorption capability of cutin (TC4) was 2.4 times higher than the nonsaponifiable fraction (TC3).The 1-naphthol interactions with tomato cuticular materials were governed by both hydrophobic-type interactions and polar (H-bonding) interactions. Removal of the wax and polysaccharide materials from the bulk tomato cuticle caused a significant increase in the sorption ability of the cuticular material.There was a linear negative trend between K_(oc) values and the amount of polysaccharides or fraction's polarities ((N O)/C);while a linear positive relationship between K_(oc) values and the content of cutin monomer (linear R~2=0.993) was observed for present in the cuticular fractions.Predominant sorbent of the hydrophobic organic compounds (HOCs) in the plant cuticular fraction was the cutin monomer,contributing to 91.7% of the total sorption of tomato bulk cuticle.