一株菲降解菌的鉴定及降解特性

从沈阳北部污水处理厂曝气池活性污泥中驯化和分离得到一株以菲为碳源的降解菌株W12,根据菌株形态和16S r DNA基因测序分析,该菌株鉴定为耳炎假单胞菌(Pseudomonas otitidis.).该菌株降解菲的最佳环境条件为:温度为30℃,p H值为7.0,摇床转速为170 r·min-1,接种量为10%,盐度为0.5%;菲初始质量浓度为1000 mg·L-1培养96 h后,降解率为65.80%,且对菲的最大耐受浓度为2000 mg·L-1;加入蛋白胨和酵母膏后,降解率分别提高到75.65%和70.85%.     

紫外诱变选育高原环境絮凝菌及其絮凝条件优化

为获得耐低温且絮凝优异的絮凝菌,对一株分离得到的絮凝菌进行紫外诱变和5℃低温胁迫培养,并对诱变优势菌进行应用条件优化.结果表明:①原始菌为蜡状芽孢杆菌（Bacillus cereus）.②紫外诱变和低温胁迫培养得到的目标诱变菌FB-5对生活污水絮凝率达75.35%,具有良好的遗传稳定性.③单因素试验确定最佳絮凝条件,即絮凝菌投加量为0.90 mL/（50 mL）、pH为7.4、助凝剂加量为1.0 mL/（50 mL）、处理时间为15 min;筛选出影响诱变菌絮凝能力最显著的3个因素为絮凝菌投加量、pH和助凝剂加量;通过Box-Behnken响应面试验得到最佳净化的应用条件,即絮凝菌投加量为0.90 mL/（50 mL）、pH为7.38、助凝剂加量为1.04 mL/（50 mL）.④验证试验确定絮凝率达97.01%,BOD₅、COD_Cr、浊度和色度去除率均达到97%以上,出水水质满足GB 18918—2002《城镇污水处理厂污染物排放标准》一级A标准.研究显示,紫外诱变联合低温胁迫技术以及单因素与响应面结合优化絮凝条件可以大幅度提高原始菌株的絮凝能力,低温诱变菌FM-5能使高寒地区城市生活污水中悬浮固体大幅降低,同时去除水体中多种污染物.      

生物降解优势菌株的选育及其含油废水降解性能研究

从被含油废水长期污染的活性污泥中筛选出能降解该废水的优势菌种。经分离纯化得到4株不同的单一菌株。通过对各菌株降解能力的考察,其中菌株H1降解能力明显高于其它菌株。经利用正交试验法实验,得到H1菌属最适生长条件为40℃温度、pH9、NaCl浓度为0．5mol／L。该菌株降解含油废水的最佳使用条件为：pH7;温度30℃-40℃;培养时间为40h;最佳菌体接种量为每100mL废水中投加2．0mL;营养液中,生长因子是必需的营养成分;而且C／N最佳比值为3：1。H1菌株的降解率高出市售菌种的降解率20％以上,有效地解决了市售菌种因针对性不强而效果欠佳的难题,有望降低生产成本。     

焦化废水中氨氮降解菌降解性能的研究

本实验从焦化废水中分离出一株高效氨氮降解菌A1,经过生理生化鉴定,初步确定为假单胞杆菌菌属。本实验对该菌种的降解性能进行研究,确定该菌种的最佳降解条件:p H8.0,温度30℃,菌种量5%(V菌悬液:V培养基),在此条件下,对200mg/L的氨氮去除率达到89.5%。     

黄原酸盐高效降解菌的降解特性研究

从矿山废水富集分离再经紫外诱变筛选出突变菌株2#,此菌株降解黄原酸盐的最佳条件为:在pH为10,温度为30 ℃,振荡速率120 r/min,有光照,接种量为10％时能高效降解黄药,降解率可达78.1％,CODG去除率达到66.2％.研究发现pb2+和Zn2+对菌株生长和降解黄药均有抑制作用,前者的抑制作用更强.当加入0.2 g/L的葡萄糖时可大大提高菌对黄原酸盐的降解率.     

α-蒎烯降解菌株的筛选及降解特性

从生产萜烯树脂的废水中分离到一株能降解α-蒎烯的菌株,通过16S r RNA序列鉴定和构建系统发育树分析,该菌株为海洋小单胞菌(Micromonospora marine),命名为HKY。研究了温度、p H值、α-蒎烯的浓度对这株菌降解α-蒎烯的影响,结果表明,菌株在25~40℃对α-蒎烯都有较好的降解,其中在30℃降解率最高,可达到90.21%。初始p H在6~8之间时降解率较高,其中p H为7时,降解率最高可达到93.43%。当α-蒎烯在100 mg/L以内时,培养48 h后没有残留,200 mg/L和300 mg/L时培养液中有残留,500 mg/L基本对该菌处于抑制状态。同时研究了该菌株的底物谱,结果表明该菌株能够降解一些分子结构较为简单或者与α-蒎烯结构相似的工业有机污染物。运用该菌处理生产萜烯树脂污水处理厂的出水,能够对COD有一定的去除作用。     

烟嘧磺隆降解菌SY-6的固定化、降解特性及其工程化应用

以海藻酸钠为载体,对烟嘧磺隆降解菌SY-6进行细胞固定化,通过单因素实验明确了菌株包埋SY-6的最优条件,并对其降解特性进行研究。结果表明,当海藻酸钠含量为4%,Ca Cl2含量为3%,固化时间为4 h,制得的固定化小球具有较强的机械强度和较好的传质性能及其对烟嘧磺隆的降解率较高。固定化细胞降解烟嘧磺隆的最佳条件为p H为7.0、温度为30℃、包菌量为5%。在不同p H或温度下,与游离细胞相比,固定化细胞有较宽泛的耐受性。将包埋的SY-6投加到序批式反应器(简称SBR)中处理烟嘧磺隆废水,结果表明,菌株SY-6在合适的使用周期内稳定性较高且对烟嘧磺隆废水降解效果较好,为烟嘧磺隆固定化细菌的工程化应用奠定了基础。     

响应面优化石油降解菌配比及降解条件研究

利用前期筛选的4株高效石油降解菌,采用响应曲面法优化混合菌配比并研究环境因素对混合菌影响,以期提高混合菌对石油的降解效率。结果表明,当石油浓度为10 g/L时,菌株Rp、Sm、Bc和Kv接种量分别为3.97%、2.00%、2.04%和2.07%时混合菌对石油降解率达最大值69.14%,验证值68.83%。此外,考察了环境因素对混合菌降解石油烃的影响,发现混合菌对盐度和p H有较大范围的适应性。其中盐度为5%时降解率仍可达40.37%;p H为7.5时降解率最高可达64.93%,p H为4时降解率最低。     

1株金霉素降解菌的筛选及基于响应面法的降解条件优化

从河南驻马店金霉素制药厂活性污泥中分离、筛选出1株金霉素降解菌,经16SrDNA鉴定为芽孢杆菌属(Bacillus sp.),命名为JMS-B01。为进一步提高该菌株的金霉素降解率,对其进行降解条件优化。采用正交试验确定显著影响金霉素降解率的3个因素(温度、金霉素浓度、接种量),通过Box-Behnken试验设计及响应面法分析确定最佳降解条件。结果表明:拟合所得的预测值与实际值相关性良好,最佳降解条件为温度34.0℃、金霉素浓度102.5 mg/L、接种量2%(体积分数),在此条件下金霉素降解率可达到96.50%。其中,温度是影响菌株JMS-B01对金霉素降解率的最重要因素之一。     

雌二醇降解菌Acinetobacter sp.DS1的筛选及降解条件优化

筛选并鉴定出1株雌激素降解菌Acinetobacter sp.命名为DS1,对DS1降解17β-雌二醇实验条件进行优化,使用响应面法(RSM)对降解过程中底物浓度、接种量、培养温度和p H值进行了分析,构建响应面模型,并对反应条件进行优化。优化最佳降解条件为:17β-雌二醇量5.10 mg/L;接种量7.5%;p H值8.7;温度27℃。并进行试验验证,结果接近,响应面模型可有效描述降解过程,为降解工程应用和设计提供技术参考。     

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.     

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.     

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.     

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.