拟除虫菊酯类农药残留降解菌产气肠杆菌的分离、鉴定及降解特性研究

采用室内培养方法,从拟除虫菊酯类农药生产车间下水道驯化污泥中分离、筛选出1株可同时降解对联苯菊酯、甲氰菊酯和氯氰菊酯的高效菌株M6R9,经鉴定为产气肠杆菌(Enterobacter aerogenes),并对其生理特性进行研究.结果表明,该菌为革兰氏阴性好氧型杆菌,大小约为长(0.8~1.9)μm、宽(0.5~1.0)μm,能够以3种菊酯农药为碳源生长.在通气、pH 7.0、温度(25~30)℃、接菌量D415 nm=0.2、农药浓度100 mg.L-1和转速180 r.min-1的环境条件下,对含有联苯菊酯、甲氰菊酯和氯氰菊酯各100 mg.L-1的混合培养基培养3 d,发现该菌对3种菊酯降解效果最好,降解率分别为55.74%、55.11%和56.96%,且降解过程满足一级动力学方程模型,降解半衰期(t1/2)分别为65.4、70.7和68.6 h.3种菊酯农药降解率与接菌量、通气量和振荡速率呈正相关.     

一株氯氰菊酯降解真菌的筛选鉴定及其降解特性研究

从雅安市售砖茶中分离筛选出一株对氯氰菊酯具有较高降解能力的真菌菌株YAT,根据其形态学及ITS序列分析,将该菌鉴定为黑曲霉(Aspergillus niger).菌株YAT在PD培养基中168 h内对50 mg·L-1氯氰菊酯的降解率为54.83％,对氯氰菊酯的降解率与菌体生物量呈正相关.动力学研究表明,菌株YAT降解氯氰菊酯的过程符合一级动力学方程,在测试的底物(氯氰菊酯)浓度、温度、pH范围内,氯氰菊酯半衰期为85.750～281.958 h,较高温度及偏碱性条件有利于氯氰菊酯的降解,底物浓度越高,其半衰期越长.此外,菌株YAT对50 mg·L-1溴氰菊酯、氰戊菊酯、氟氯氰菊酯和氯菊酯的120 h降解率分别为27.53％、58.00％、53.23％和25.34％.     

缺陷假单胞菌M5R14粗酶液降解拟除虫菊酯类农药特性初探

以分离于杭州农药厂污水排放污泥的1株可降解多种拟除虫菊酯类农药的菌株--缺陷假单胞菌M5R14为对象,采用直接离心分离提取和超声波破碎提取的方法获得菌株M5R14的胞外和胞内粗酶,初步探讨了该菌株粗酶液降解拟除虫菊酯类农药的特性.研究表明该菌株降解拟除虫菊酯类农药的酶为胞内酶,粗酶液对联苯菊酯、甲氰菊酯和氯氰菊酯3种拟...     

太湖及辽河流域表层沉积物中拟除虫菊酯的浓度水平及毒性评估

对辽河和太湖流域表层沉积物中8种常见的拟除虫菊酯:联苯菊酯、高效氯氟氰菊酯、氯菊酯、甲氰菊酯、氟氯氰菊酯、氯氰菊酯、氰戊菊酯、溴氰菊酯进行分析测定.结果显示,太湖流域中拟除虫菊酯含量范围为0.02～2.91ng·g-1(干重,下同),平均含量为0.63ng.g-1,处于相对较低的水平;辽河流域中的含量相对较高,其范围是0.14～19.6ng·g-1,平均含量为6.20ng·g-1.采用毒性单元TU(选用钩虾Hyallela Azteca的LC50)对拟除虫菊酯毒性风险进行估算,太湖流域∑TU值范围是0～0.40,相比之下,辽河流域的∑TU值高出很多,范围在0.10～1.73之间,其中有3个点位的∑TU值大于1,表明其具有一定的生态风险,其毒性主要来源于氯氰菊酯和氰戊菊酯.     

一株1,2,3,4-四氯苯降解菌的筛选和降解特性研究

从广东某化学试剂厂周边土壤中筛选、分离并纯化出一株能有效降解1,2,3,4-四氯苯(1,2,3,4-TeCB)的菌株,命名为L-1号菌.探索不同接种量(2%;3%;5%)菌株对25 mg·L-1的1,2,3,4-四氯苯模拟废水化学需氧量(COD)、氯离子释放浓度、1,2,3,4-四氯苯降解效果影响,并结合气相质谱联用仪(GC-MS)工作站分析菌株对1,2,3,4-TeCB降解产物.结果表明:经16S rDNA鉴定该菌株为枯草芽孢杆菌;菌株接种量为5%对1,2,3,4-TeCB降解效果最佳,在pH=7.0、温度30℃,摇床转速110 r·min-1条件下反应7d,COD降解率达62.71%;氯离子释放浓度4.98mg·L-1;四氯苯降解量达20.47 mg·L-1.通过对L-1号菌降解1,2,3,4-TeCB产物分析,发现L-1号菌对1,2,3,4-TeCB的降解是通过先开环后脱氯的途径实现的.     

两株毒死蜱降解细菌的分离鉴定及其降解特性

采用富集培养的方法,从农药生产企业的废水处理系统中分离驯化出两株能够降解毒死蜱的菌株HY-2与HY-4,在形态特征和生理生化分析的基础上,又对其16S rDNA序列进行了分析,并研究了其对毒死蜱和其它有机磷农药的降解特性.结果表明,2个菌株均属蜡状芽孢杆菌(Bacillus cerens),HY-4对毒死蜱的降解能力大于HY-2.2个菌株降解毒死蜱的适宜条件为:外加葡萄糖浓度3g·L-1,培养温度35℃,初始pH值8.0,毒死蜱初始浓度80mg·L-1,接种量20%(体积比,菌体密度:稀释到所配菌悬母液(OD600=2)的0.5倍).酵母膏含量对降解的影响表明,当添加3g·L-1的葡萄糖时,最适的酵母膏含量为1g·L-1,而不添加葡萄糖时,最适的酵母膏含量为5g·L-1.2个菌株对甲基对硫磷的降解研究发现,HY-2和HY-4对初始浓度为100mg·L-1的甲基对硫磷60h的降解率分别达64.7%和88.7%;然而,2个菌株对初始浓度为100mg·L-1的三唑磷60h的降解率仅为13.3%-14.2%,降解率较低.     

低营养水体中芽孢杆菌降解有机氮的研究

芽孢杆菌具有降解有机氮的功能,但在养殖水体等低营养水体中,其降解效果可能受到影响.为研究低营养水体中有机氮的降解情况,通过模拟凡纳滨对虾中间培育过程配制低营养水体,分别接种芽孢杆菌NT9和YB3(NT9水体和YB3水体),然后研究水体中微生物的生长与有机氮的降解情况,并构建数学模型进行分析.结果显示,起始接种量为10×10~5 cfu·mL~(-1)时,NT9水体总菌量呈下降趋势,平均为(3.46×10~5±2.39×10~5) cfu·mL~(-1),YB3水体总菌量则上升到(25.43×10~5±8.84×10~5) cfu·mL~(-1),但均高于未接种的对照水体.NT9水体和YB3水体的有机氮降解率显著高于对照水体(p0.05),分别提高50.28%和119.41%,降解速率也分别提高65.22%和121.74%.对照水体、NT9水体和YB3水体单位菌量的有机氮降解效率分别为1.238、1.649和1.904 mg·L~(-1),降解模型分别为y=-6.40+1.39x_1+1.45x_2、y=2.11+8.21x_3-0.64x_4-1.26x_1x_3-0.32x_2x_4和y=1.73+6.11x_2(x_1、x_2、x_3、x_4分别表示总菌量、总菌增量、有机氮含量和时间).研究表明,在低营养水体中接种芽孢杆菌有利于有机氮的降解,但不同的菌株具有不同的降解模式,菌株YB3为能够适应低营养水平、增殖能力较强的菌株,可以更有效地促进有机氮的降解,提高降解效率.     

拟除虫菊酯类杀虫剂的酶促降解

测定了降解菌Ａｌｃａｌｉｇｅｎｅｓ ｓｐ．ＹＦ １１粗酶液对拟除虫菊酯杀虫剂的降解性能,粗酶液降解杀灭菊酯、甲氰菊酯、氯氯菊酯、溴氰菊酯氯菊酯、三氟氯氰菊酯的最适ｐＨ分别为８．０、８．５、８．０、８．０、９．０、７．５;米氏常数分别为４１．４、１３６．８、６５．４、２２２．８、５．２、８．６７ｎｍｏｌ／ｍｌ;最适温度为３２．５℃,该粗酶液在ｐＨ５．０－１０．５均具有降解活性。     

2株降解菲的植物内生细菌筛选及其降解特性

为了获得具有菲降解特性的植物内生细菌,通过选择性富集培养,本研究从多环芳烃污染区植物体内分离得到2株能够降解液体培养基中高浓度菲(200 mg·L-1)的植物内生细菌(菌株P1和P3).经形态观察、生理生化特征鉴定和16S rDNA序列同源性分析,分别将菌株P1和P3鉴定为寡养单胞菌属(Stenotrophomonas sp.)和假单胞菌属(Pseudomonas sp.)的细菌.菌株P1和P3均为好氧生长,28℃、150 r·min-1摇床培养7 d,2株菌对无机盐培养基中菲(100 mg·L-1)的降解率均高于90%.条件实验表明,温度20～30℃,pH 6.0～8.0,盐含量0%～4%,装液量10～30 mL(100 mL三角瓶)2菌株生长良好且对菲降解率高于70%.其最适生长和降解温度为30℃,pH为7.0,盐含量≤4%,装液量≤30 mL.综合比较2株菌对菲的降解特性,P1菌株高温耐受性稍强,而P3菌株对环境pH改变和缺氧的耐受性稍强.     

农药微生物降解性与农药疏水性参数间的相关性

在纯培养条件下，产碱菌属广说性降解菌ＹＦ１１对对硫磷，甲基对硫磷，杀螟松，氰戊菊酯，溴氰菊酯，甲氰菊酯，氯氰菊酯，三氟氯氰菊酯，氯菊酯的降解速率的差异主要是由各杀虫剂的疏水性参数的不同引起的，两之间存在较好的相关性：ｌｏｇＫ＝２．２１－０．２６ｌｏｇＫｏｗ，ｒ＝０．９７３９。     

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

ＲｅｍｏｖａｌｏｆｈｅａｖｙｍｅｔａｌｓｆｒｏｍｓｅｗａｇｅｓｌｕｄｇｅｂｙｌｏｗｃｏｓｔｉｎｇｃｈｅｍｉｃａｌｍｅｔｈｏｄａｎｄｒｅｃｙｃｌｉｎｇｉｎａｇｒｉｃｕｌｔｕｒｅＷｕＱｉｔａｎｇ，ＮｙｉｒａｎｄｅｇｅＰａｓｃａｓｉｅ，ＭｏＣｅｈｕｉＦ...     

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.