一株酸性大红3R脱色菌的分离、鉴定及其脱色性能研究

通过浓度梯度驯化,从厌氧污泥中分离到一株酸性大红3R脱色菌M3,根据其形态学特征及16S rRNA基因序列分析,鉴定该厌氧菌株为阴沟肠杆菌(Enterobacter cloacae).同时,研究了该菌株在厌氧静置条件下对酸性大红3R的最适脱色条件.结果表明,菌株脱色的最佳营养源为酵母粉,最适宜脱色条件为酵母粉浓度1％、pH =6.5、温度35 ℃、盐度小于4％,在此条件下,对浓度为100 mg·L-1酸性大红3R溶液厌氧静置培养72 h,脱色率达到98％以上.菌株耐盐性良好,在6％ ～8％的盐度下具有较好的脱色效果.分光光谱表明,菌株脱色机制主要为生物降解脱色.毒性试验表明,脱色后的酸性大红3R毒性明显降低.     

Trametes sp.LS-10C产漆酶发酵培养基优化及其漆酶对偶氮染料的脱色性能

为了提高菌株Trametes sp.LS-10C的漆酶产量并初步研究该酶对偶氮染料的脱色性能,本文通过单因素实验及均匀设计对菌株Trametes sp.LS-10C产漆酶培养基进行了优化,获得最优培养基配方为:葡萄糖14.4 g·L~(-1)、豆渣10.1 g·L~(-1)、麸皮100 g·L~(-1)、NH_4Cl_3g·L~(-1)、KH2PO41.4g·L~(-1)、CuSO_4’5H2O 1 g·L~(-1)、NaCl 1 g·L~(-1)、MgSO_40.8 g·L~(-1)、CaCl_20.5 g·L~(-1).菌株发酵至8 d时漆酶产量为595.15 U·m L~(-1),约是优化前的35.66倍.进一步研究表明,该漆酶-介体系统对酸性红GR、酸性蓝40和酸性铬蓝K等3种偶氮染料脱色10 h后脱色率分别高达96.86%、91.28%和86.31%.相比公开发表的文献,本研究所报道的Trametes sp.LS-10C利用优化培养基发酵产漆酶达到了较高水平,具有酶活力高和发酵时间短等特征,且该漆酶在处理偶氮染料废水脱色领域中具有进一步研发和应用价值.     

Citrobacter sp. LW-3对偶氮染料甲基橙的降解脱色特性研究

从长期受染料污染的土壤中分离出一株能对甲基橙高效降解脱色的柠檬酸盐杆菌Citrobacter sp.LW-3.菌株LW-3在添加了0.5%(g/100 mL)葡萄糖的MSM-1培养基中,16 h使100 mg·L-1甲基橙降解掉90.56%.该菌株对甲基橙降解脱色可在完全好氧条件下,脱色与木质素过氧化物酶、NADH-DCIP原酶、核黄素还原酶有关.菌株LW-3降解甲基橙的适宜温度较广,20~40℃24 h均能使超过80%的甲基橙降解脱色,适宜pH范围为6.0~8.0.通气量对菌株LW-3影响较小,24 h各装液量组(25~200 mL)甲基橙均能降解近90%.同时,菌株LW-3具有较广的染料降解谱,3 d内可使20 mg·L-1另3种偶氮染料、两种三苯甲烷类染料和一种蒽醌类染料很好脱色.菌株LW-3在染料废水的生物处理方面有较强的实际应用价值.     

耐盐菌对偶氮染料酸性红B的脱色研究

从食品加工厂含盐污水处理车间的活性污泥中分离出对偶氮染料具有脱色活性的耐盐菌体,研究了不同pH值、脱色温度、接菌量以及装液量等条件对耐盐菌体降解酸性红B的影响,并设计正交实验,获得其降解模拟偶氮染料废水的最佳反应条件.研究结果表明,耐盐菌体TAS在35℃、pH为8、装液量为50mL及接菌量为8%(体积分数)的条件下,反应10h后对浓度为50mg·L-1的酸性红B模拟废水的脱色率达91%.     

醌介导染料脱色菌株的分离鉴定及特性

实验分离获得1株能够利用醌化合物使磺酸化偶氮染料脱色的菌株JL,通过形态特征、16S rDNA与16S-23S区间序列分析表明,该菌株为蜡状芽孢杆菌(命名为Bacillus cereus JL).菌株JL使酸性大红3R脱色的最佳条件为葡萄糖浓度1g/L, pH值为5~7,温度30℃,接种量0.25g/L.蒽醌-2-磺酸(AQS)、蒽醌-2,6-二磺酸(AQDS)和2-羟基-1,4-萘醌(Lawsone)均能显著提高酸性大红3R的脱色速率,其中AQS的促进作用最为明显.研究发现, 0.1mmol/L AQS能够使菌株JL对2.0mmol/L酸性大红3R保持较高的脱色速率,而且能使多种偶氮染料脱色,表现出较好的底物广谱性.利用高效液相色谱-质谱鉴定了AQS介导的酸性大红3R脱色产物,表明酸性大红3R的偶氮键发生断裂, AQS在这一过程中仅起到电子传递的作用.     

Enterobacter sp. CV-v对甲基橙的脱色特性与条件优化

从浙江温州分离筛选到1株甲基橙高效脱色菌株CV-v,经16S rRNA基因序列分析表明,该菌株属于Enterobacter sp.属.单因素实验结果表明:当pH值在5.0~9.0之间时,培养48h以后,该菌株对甲基橙的脱色率均在80%以上;脱色的最适温度范围为30~40℃之间;所测碳源中的葡萄糖和麦芽糖、氮源中的牛肉膏和酵母粉对脱色的促进效果最为显著,且Ca2+和Mg2+也对脱色有显著的促进效应;此外,当接种量达到7%(V:V)以后,48h的脱色率即可达近100%.响应面设计实验结果显示,该菌株对甲基橙脱色的最优操作条件为:pH 8.0,葡萄糖1.7g/L,酵母粉3.0g/L,氯化镁3.0mmol/L及培养温度35.9℃.验证实验结果表明在最优条件下,该菌株在10h内对甲基橙的脱色率可达88.0%.总体而言,该菌株在偶氮染料脱色中的应用潜能较大.     

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%.     

酵母Candida krusei对合成染料的脱色

通过筛选实验,从土壤中新分离到1株对活性艳红K-2BP具有明显脱色效果的酵母菌株Y-G-1,经鉴定为克鲁斯假丝酵母Candida krusei.该菌株对含活性艳红K-2BP起始浓度为200mg/L的培养基,最大脱色率为99%,达到最大脱色率的时间是12h.克鲁斯假丝酵母的最佳接种量应不低于5%(体积分数),培养基最适pH在4～9之间,氮源(NH₄)₂SO₄浓度不低于0.1%,相对应的碳源葡萄糖浓度不低于0.5%.对脱色机理的研究表明,该菌株对活性艳红K-2BP的去除属于降解脱色.此外,该菌株对另外9种染料(50mg/L)的脱色率在62%～96%之间.其中,对偶氮染料弱酸艳红B、活性黑KN-B和活性红M-3BE的脱色率都达到了90%以上,对三苯甲烷染料(酸性媒介漂蓝B)的脱色率达到了93%.表明克鲁斯假丝酵母在染料废水的处理上可能具有较好地应用潜能.     

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.     

酸性偶氮染料还原产物强化偶氮染料生物脱色

考察醌还原菌群利用酸性偶氮染料的生物还原产物对偶氮染料生物脱色的强化作用.结果表明,酸性红B 等酸性偶氮染料的还原产物作为氧化还原介体可以强化多种偶氮染料的生物脱色.当酸性红B 还原产物浓度为0.2mol/L 时,其对活性艳红KE-3B 最佳脱色pH 值为7~11,在4~40℃范围内脱色率随温度上升而提高.根据高效液相色谱-质谱对酸性红B 还原产物的分析结果,推测起氧化还原介体作用的物质是2-氨基-4-磺酸基-1-萘酚     

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.     

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.     

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.