酵母菌T-2对蒽醌染料的脱色研究

从温州市上桥一家染料厂的污泥中筛选分离到一株染料脱色优势酵母菌T 2。经初步鉴定为红酵母属 (RhodotorulaHarri sonsp .)。并研究了该菌在不同温度、pH、培养时间、染料浓度、接种量等条件下对染料脱色的情况。同时 ,还研究了不同碳源、氮源等营养条件下对活性艳蓝KN R脱色的情况。实验结果表明 :在 30℃～ 35℃温度范围内T 2菌对活性艳蓝KN R的脱色率为 89.9%～ 95.1 % ,最佳脱色温度为 35℃ ,pH值在 2 .0～ 7.0范围内T 2对活性艳蓝KN R的脱色率为 81 .7%～ 92 .6 % ,最佳脱色 pH为 3.0 ,碳源、氮源、接种量及染料浓度对其脱色率均有影响     

东方伊萨酵母YP-1对染料活性艳红K-2BP的脱色

利用含染料的选择性培养基从土壤中分离出一株对活性艳红K-2BP有明显脱色效果的酵母菌株YP-1,经鉴定为东方伊萨酵母Issatchenkia orientalis.结果表明,该酵母菌对￡400mg/L的活性艳红K-2BP有较好的脱色效果.对于活性艳红K-2BP起始浓度为100mg/L的培养基,该菌株可在12h达到99%以上的最大脱色率,其最佳接种量为10%(体积分数),最适pH值在3~9之间,氮源(NH4)2SO4的浓度30.02%,碳源葡萄糖的浓度30.2%.脱色机理研究结果表明,该酵母对活性艳红K-2BP的去除是先吸附后生物降解.此外,该菌株对初始浓度为200mg/L的偶氮染料活性黑KN-B的脱色率也可达99.5%.     

3株真菌对活性艳蓝KN-R的脱色条件

从受污染土壤中筛选出具有广谱脱色的优势菌17株,并进一步扩大染料范围,筛选出3株对偶氮、蒽醌、三苯甲烷染料均有较好脱色效果的优势菌株. 3株菌分别为青霉属(菌Ⅰ、菌Ⅱ)和头孢霉属(菌Ⅲ)的真菌;以染料配水为例,探讨了pH值、温度、碳源、氮源因素对菌株脱色的影响;并进行了实际废水的脱色实验,结果表明该菌在pH值5～9, 温度18～37℃的区间内,且外加葡萄糖提供0.5%碳源时对染料废水的脱色率可达70%. 因此对处理染料污染废水具有较好的应用前景.     

高效产絮凝剂Pseudomonas alcaligenes培养条件优化及应用研究

以筛选出的一株高效絮凝剂产生菌产碱假单胞菌PS-25为试验菌种,采用单因素试验方法确定最佳碳源为葡萄糖,最佳氮源为黄豆饼粉。采用正交试验设计方法,对该菌产絮凝剂的培养条件进行优化研究,得到最佳培养条件是:碳源为葡萄糖(20.0g/L)、氮源为黄豆饼粉(4.0g/L)、培养基初始pH值为6.5、培养温度为30℃、接种量5%(V/V)、通气量为160r/min。在最佳培养条件下,PS-25产生的絮凝剂对高岭土悬浊液絮凝率达95.77%,且对多种实际废水都有较好的净化效果,其中对浊度的去除率均在91%以上,对色度的去除率均在80%以上,对废水中COD的去除率为54.25%～90.33%。     

P.phragmitetus发酵条件优化及其所产絮凝剂的应用研究

在LB培养基的基础上,对P.phragmitetus的生长特性进行研究,发现P.phragmitetus所产絮凝剂的表达与菌体呈正相关,且倾向于在碱性条件(pH=7.5)下生长和产生絮凝剂。选择察氏培养基为基础发酵培养基,发现P.phragmitetus产絮凝剂最佳碳源为葡萄糖,氮源为蛋白胨,最适碳氮比为4∶1。以该菌所产絮凝剂粗品和脱色剂双氰胺甲醛树脂对模拟的活性染料废水进行联合处理。结果表明,微生物絮凝剂投加量为40μg/mL,脱色剂用量为100μg/mL时,脱色率达到90%。相对于传统化学处理工艺,该联合处理工艺所需脱色剂和絮凝剂用量都更少。     

复合菌利用酱油废液制备生物絮凝剂及其絮凝特性分析

利用酱油曲霉(Aspergillus sojae)和毕赤酵母(Pichia membranifaciens)复合菌以酱油酿造废液作为培养基生产生物絮凝剂,研究了外加碳源、外加氮源、培养基的初始pH值以及酱油酿造废液浓度等因素对絮凝剂产量及絮凝剂活性的影响,并考察了所制备絮凝剂的絮凝特性.结果表明,酱油酿造废液稀释1倍,加入5g/L乙醇作为补充碳源,不需添加氮源,调节pH值至5.0左右,培养30h,生物絮凝剂的产量可达到5.92g/L.实验制备的生物絮凝剂具有较好的絮凝效果,对高岭土悬浊液的去除率达到98.3%;对酱油废水、酿酒废水、生活污水、乳品废水等实际废水的浊度的去除率78.2%~92.3%,COD去除率为64.2%~85.2%.     

降解酸性蓝B的镰刀菌(Fusarium sp.)HJ01的分离和降解特性研究

从膨润土中分离出一株对酸性蓝B具有降解效果的镰刀菌(Fusariumsp.)HJ01,研究了其生长特征和动力学模型模拟、降解酶的性质以及对酸性蓝B的脱色效果.结果表明:菌株生长的适宜条件为:pH范围为6~7,蔗糖为碳源,NH4Cl为氮源.菌体生长符合改进的logistic law模型.SDS-PAGE分析HJ01菌株所产漆酶的相对分子量约为66×103,在低氮条件下漆酶活性高达431~812U·mg~(-1)·s~(-1).菌体培养96h后加入到含100mg·L~(-1)酸性蓝B的培养基中,再培养96h,酸性蓝B的脱色率达100%.紫外光谱分析表明,酸性蓝B的发色基团蒽醌环被漆酶破坏.     

沼泽红假单胞菌对活性紫KBR脱色的研究

研究了驯化后的沼泽红假单胞菌(Rhodopseudomonaspalustris)N菌株的生长细胞在不同条件下对活性紫KBR的脱色效果.结果表明,该菌株生长细胞脱色的最佳条件为温度25～30℃,pH值为7,厌氧条件下的脱色率远远高于好氧条件下的脱色率.染料作为该菌株唯一的碳源和能源脱色时,脱色率与细胞浓度呈极显著相关,细菌脱色比活率保持在较恒定的水平.此外,生长细胞脱色过程中对菌体生长的测定表明,该菌株生长的指数期明显滞后于染料脱色的指数期.     

染料活性艳红X-3B的微生物脱色研究

实验研究了以葡萄糖为外加碳源、由酵母膏提供氮源时不同的影响条件下紫色非硫光合细菌的混合菌种对染料活性艳红X-3B的脱色效果。结果表明,随着葡萄糖浓度的增大,脱色率反而下降;酵母膏的浓度对脱色效果影响很大,它在脱色过程中起到氮平衡的作用;当葡萄糖和酵母膏的浓度分别为0.125%和0.3%时,X-3B的脱色效果最佳。所选实验条件下,染料浓度在500mg/L以下X-3B脱色效果很好,对菌种没有明显的抑制作用;染料浓度在500～1000mg/L之间变化时,虽然对菌种有抑制作用,染料最终仍能完全脱色。温度在30℃时,pH在3～10范围内染料均有较好的脱色效果。     

高效产絮凝剂荧光假单胞菌C-2培养条件优化应用

以筛选出的一株高效产絮凝剂荧光假单胞菌C-2为菌种,采用单因素试验方法确定最佳碳源为葡萄糖,最佳氮源为黄豆饼粉。采用正交试验设计方法,对该菌产絮凝剂的培养条件进行优化研究,结果表明:菌株产絮凝剂的最佳培养条件是碳源为葡萄糖(20.0g/L),氮源为黄豆饼粉(4.0g/L),培养温度为30℃,培养基初始pH值为8.0,通气量为160r/min,在此培养条件下,对高岭土悬浊液絮凝率达93.52%。最佳培养条件下产生的絮凝剂用于多种实际废水的净化,其中对色度和浊度的去除率均在80%以上,对COD的去除率为52.67%～85.22%。     

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.