溴氰菊酯降解菌的筛选及降解特性研究
|
摘要
生物修复对降解污染基质中的农药是一种有效的环境污染修复方法。从自然界中筛选出能够降解特定污染物的高效菌种,有针对性地投加到受污染环境中的生物强化技术,能够快速提供具有降解作用的微生物,在环境的农药污染治理中显示出巨大的潜力。溴氰菊酯是拟除虫菊酯类农药,这类农药为神经类毒物,属高毒性物质,其应用非常广泛,是杀虫剂中的第三大类。该类农药残留已成为目前农产品的主要农药残留类型之一,在污染环境的同时严重危害人类的健康。微生物在农药残留降解过程中起着重要的作用,微生物降解技术已成为去除农药残留的绿色生产技术。因此,积极开展溴氰菊酯降解菌的筛选工作有着重要意义。
本论文目标是寻找能够降解溴氰菊酯农药的高效降解菌。从长期使用溴氰菊酯的菜园土壤和过期的农药中筛选出能够以溴氰菊酯为唯一碳源生长的降解菌株,并研究其降解效率、降解特性及其农药降解谱。此外,通过室内盆栽试验研究其最优化降解条件。
最终分离得到两株溴氰菊酯降解菌(编号为X09、X24)。经Biolog自动微生物分析系统鉴定结果为:菌株X09为阴沟肠杆菌Enterobacter cloacae,菌株X20为荧光假单胞菌Pseudomonas fluorescens。两种降解菌在溴氰菊酯为唯一碳源的选择性培养基中生长良好,在pH7.0、添加50mg/L溴氰菊酯为唯一碳源的基础培养基中,30℃振荡培养,接种7天后,对溴氰菊酯的降解率分别达62.82%、43.25%。菌株X09、X20对溴氰菊酯的最大耐受浓度为800mg/L。
降解菌株的最优化降解条件为温度30℃、pH7.0、接种量25%和培养时间96h。其中菌株X09在该条件下降解率高达65%以上;菌株X20在外加少量碳源如淀粉、葡萄糖、牛肉膏等,可以显著提高其降解能力,达到69.3%。此外,两种降解菌对其他拟除虫菊酯类农药也有一定的降解能力,对氯氰菊酯、甲氰菊酯、氰戊菊酯的平均降解率分别为53.0%,49.9.%,56.7%。
室内盆栽正交试验统计结果表明,各种因素对菌株降解能力影响大小依次为菌株种类、处理时间、喷菌量、处理方式及施药量。统计分析结果为,最优化降解条件为菌株X24,施药量为10mg/L,喷菌量为5mL/株,处理方式为菌液与农药混和喷施以及处理时间为7天。另外,菌液和农药混和喷施的处理方法取得了较好的降解效果,为溴氰菊酯降解菌剂的研究及微生物修复农药残留问题提供了一定的数据参考。
Bioremediation is an environmentally-benign method for degrading insecticides from contaminated substrates.Bacteria that can effectively degrade pesticide compound had been isolated from natural environment,they can be pour into contaminated system and obviously enhanced the effect,therefore,they have extraordinary potential in bioremediation.Pyrethroids are neural toxicum substance and is very harmful to human health.Its origin existed wildly in our life.Synthetic Pyrethroid insecticides is the third largest class of insecticides.The residue of this kind of insecticides has become one of the most dominant insecticide residues in farm produce.And microorganism has important function on degrading pesticide residues.Therefore, the technology of microbial degradation has turned into the organic production technology to remove pesticide residues.
In this text,our goal was to find bacteria that could degrade the Pyrethroids that are used in agriculture production.We isolated bacteria from pesticide-treated contaminated soil from vegetable garden.Initial screening was on enrichment media,which subsequent sequential isolation on minimal media containing the pesticides as sole C source.
As a result,two strains which grew particularly well(designated X09 and X20) was Enterobacter cloacae and Pseudomonas fluorescens respectively,identified with the Biolog whole-auto-microbial analysis system.We then used gas chromatography HP6890 to measure the rates of disappearance of the deltamethrin from pH 7.0,liquid cultures maintained aerobically at 30℃.Other than 50mg/L of the pesticides,the media were carbon flee.Seven days after inoculation with X09 and X20the concentrations of the deltamethrin decreased was 62.82%and 43.25%respectively.
The optimal conditions for the aerobic degradation of deltamethrin by two strains should be Nutrition Agar substrate with the content of 50mg/L Deltamathrin,at 30℃,with pH 7.0 and 96 hours and inoculation 25%.Growth measured as optical density measured the rate of disappearance of other pesticides,we showed that two strains also degrade the Cypermethrin, Fenpropathrin and Fenvalerate,53.9%,49.9%and56.7%being average degraded respectively, after 7 days.Preliminary observation of the stains,they were all grew negative reaction,we further indentify the physiological and biochemical of the strains.Since X09 Enterobacter cloacae and X20 Pseudomonas fluorescens can degrade a broad spectrum pesticides,it should be able to use it in a bioremediation process to minimize contamination of the product and the environment during primary products cultivation.
From the pot-culture orthogonal experiment statistical analysis result,we can see the optimal conditions for the degradation of deltamethrin were that,strain-X20,each plant was sprayed 10mg/L Deltamethrin,5mL culture liquid,spraying bacteria culture liquid mixed with deltamethrin and kept for 7 days.In this paper,through nearly two years of laboratory work,we isolated Deltamethrin-degrading bacteria.Study on its degrading characteristics,then we simulated environmental conditions,explore they degradation effect.We work for microbial remediation of pesticide residues,then provide data basis.
本论文目标是寻找能够降解溴氰菊酯农药的高效降解菌。从长期使用溴氰菊酯的菜园土壤和过期的农药中筛选出能够以溴氰菊酯为唯一碳源生长的降解菌株,并研究其降解效率、降解特性及其农药降解谱。此外,通过室内盆栽试验研究其最优化降解条件。
最终分离得到两株溴氰菊酯降解菌(编号为X09、X24)。经Biolog自动微生物分析系统鉴定结果为:菌株X09为阴沟肠杆菌Enterobacter cloacae,菌株X20为荧光假单胞菌Pseudomonas fluorescens。两种降解菌在溴氰菊酯为唯一碳源的选择性培养基中生长良好,在pH7.0、添加50mg/L溴氰菊酯为唯一碳源的基础培养基中,30℃振荡培养,接种7天后,对溴氰菊酯的降解率分别达62.82%、43.25%。菌株X09、X20对溴氰菊酯的最大耐受浓度为800mg/L。
降解菌株的最优化降解条件为温度30℃、pH7.0、接种量25%和培养时间96h。其中菌株X09在该条件下降解率高达65%以上;菌株X20在外加少量碳源如淀粉、葡萄糖、牛肉膏等,可以显著提高其降解能力,达到69.3%。此外,两种降解菌对其他拟除虫菊酯类农药也有一定的降解能力,对氯氰菊酯、甲氰菊酯、氰戊菊酯的平均降解率分别为53.0%,49.9.%,56.7%。
室内盆栽正交试验统计结果表明,各种因素对菌株降解能力影响大小依次为菌株种类、处理时间、喷菌量、处理方式及施药量。统计分析结果为,最优化降解条件为菌株X24,施药量为10mg/L,喷菌量为5mL/株,处理方式为菌液与农药混和喷施以及处理时间为7天。另外,菌液和农药混和喷施的处理方法取得了较好的降解效果,为溴氰菊酯降解菌剂的研究及微生物修复农药残留问题提供了一定的数据参考。
Bioremediation is an environmentally-benign method for degrading insecticides from contaminated substrates.Bacteria that can effectively degrade pesticide compound had been isolated from natural environment,they can be pour into contaminated system and obviously enhanced the effect,therefore,they have extraordinary potential in bioremediation.Pyrethroids are neural toxicum substance and is very harmful to human health.Its origin existed wildly in our life.Synthetic Pyrethroid insecticides is the third largest class of insecticides.The residue of this kind of insecticides has become one of the most dominant insecticide residues in farm produce.And microorganism has important function on degrading pesticide residues.Therefore, the technology of microbial degradation has turned into the organic production technology to remove pesticide residues.
In this text,our goal was to find bacteria that could degrade the Pyrethroids that are used in agriculture production.We isolated bacteria from pesticide-treated contaminated soil from vegetable garden.Initial screening was on enrichment media,which subsequent sequential isolation on minimal media containing the pesticides as sole C source.
As a result,two strains which grew particularly well(designated X09 and X20) was Enterobacter cloacae and Pseudomonas fluorescens respectively,identified with the Biolog whole-auto-microbial analysis system.We then used gas chromatography HP6890 to measure the rates of disappearance of the deltamethrin from pH 7.0,liquid cultures maintained aerobically at 30℃.Other than 50mg/L of the pesticides,the media were carbon flee.Seven days after inoculation with X09 and X20the concentrations of the deltamethrin decreased was 62.82%and 43.25%respectively.
The optimal conditions for the aerobic degradation of deltamethrin by two strains should be Nutrition Agar substrate with the content of 50mg/L Deltamathrin,at 30℃,with pH 7.0 and 96 hours and inoculation 25%.Growth measured as optical density measured the rate of disappearance of other pesticides,we showed that two strains also degrade the Cypermethrin, Fenpropathrin and Fenvalerate,53.9%,49.9%and56.7%being average degraded respectively, after 7 days.Preliminary observation of the stains,they were all grew negative reaction,we further indentify the physiological and biochemical of the strains.Since X09 Enterobacter cloacae and X20 Pseudomonas fluorescens can degrade a broad spectrum pesticides,it should be able to use it in a bioremediation process to minimize contamination of the product and the environment during primary products cultivation.
From the pot-culture orthogonal experiment statistical analysis result,we can see the optimal conditions for the degradation of deltamethrin were that,strain-X20,each plant was sprayed 10mg/L Deltamethrin,5mL culture liquid,spraying bacteria culture liquid mixed with deltamethrin and kept for 7 days.In this paper,through nearly two years of laboratory work,we isolated Deltamethrin-degrading bacteria.Study on its degrading characteristics,then we simulated environmental conditions,explore they degradation effect.We work for microbial remediation of pesticide residues,then provide data basis.
引文
[1]钱暑强,刘铮.污染土壤修复技术介绍[J].化工进展,2000,21(4):10-12
[2]李凯峰,温青,石汕.污染土壤的生物修复[J].化学工程师,2002,93(6):52-53
[3]杨国栋.污染土壤微生物修复技术主要研究内容和方法[J].农业环境保护,2001,20(4):286-288
[4]Bento F M,Camargo F A,Okeke B C,et al.Comparative bioremediation of soils contaminated with diesel oil by natural attenuation,biostimulation and bioaugmentation[J].Bioresource technology,2005,96:1049-1055
[5]Takeuchi M,Nanba K,Iwamoto,etal.In situbioremediation of a cisdichloroethylene utilizing methanerich groundwater from an uneontaminatedaquifer[J].Water research,2005,2438-2444
[6]滕应,黄昌勇.重金属污染土壤的微生物生态效应及修复研究进展[J].土壤与环境,2002,11(1):85-89
[7]张春桂,许华夏,姜晴楠.污染土壤生物恢复技术[J].生态学杂志,1997,18(4):52-58
[8]Herkovits J,Perez C,Herkovits F D.Ecotoxicological studies of environmental samples from Buenos Airesarea using a standardized amphibian embryo toxicitytest(AMPHITOX)[J].Environmental Pollution,2002,116(1):177-183
[9]霍美玲.葛根素对溴氰菊酯急性中毒的实验治疗[J].卫生毒理学杂志,1988,2(4):124
[10]耿文奎,杨玉英.溴氰菊酯毒性研究现状[J]广西预防医学,1996,2(5):308-310
[11]陈天铎,关育芳.溴氰菊酯对微小牛蜱的敏感度和对雌蜱产卵的影响的研究[J].中国兽医寄生虫病,1994,2(1):6-9
[12]王明学,扶庆,周至刚,等.溴氰菊酯对草鱼早期发育阶段的毒性效应[J].渔业,2000,20(6):39-40
[13]张毓琪,陈叙龙.环境生物毒理学[M].天津:天津大学出版社,1993
[14]贺全仁,聂星湖.拟除虫菊酯类农药的临床毒理学研究现状[J].中国工业医学杂志,1997,10(1):46-48
[15]胡春容.拟除虫菊酯农药的毒性研究进展[J].毒理学杂志,河北,唐山:2005,19(3):239-241
[16]霍美玲.葛根素对溴氰菊酯急性中毒的实验治疗[J].卫生毒理学杂志,1988,2(4):124
[17]Papadopulou,Mourkidou El Analusis ofEstablished Pyrethroids[J].Inseeticidel Residue Rev,1983,89:79-208
[18]刘世亮,骆永明,丁克强,等.土壤中有机污染物的植物修复研究进展[J].土壤,2003,35(3):187-192
[19]董元华,张桃林.基于农产品质量安全的土壤资源管理与可持续利用[J].土壤,2003,35(3):182-186
[20]Liao M,Xie XM,Subhani A.Combined effect of nutrientand pest management on soil ecological quality in hybridrice double-cropping system[J].Pedosphere,2003,13(2):129-138
[21]农业部农药检定所.新编农药手册[M].北京:中国农业出版社,1989
[22]刘均,何燕,蔡克健,溴氰菊酯残留物的分析方法研究[J].云南环境科学,1995,14(2):56-66
[23]Dean N.Advancing research for bioremediation[J].Environment Problem,1992,3(7):19-25
[24]ELROYAC.The Rhizosphere[M].Berlin:Spring-Vedag,1986
[25]Stager C E,Davis J R1 Automated Systems for identification of microorganisms[J].Clinical Microbiology Reviews,1992,5:302-307
[26]Kim C H,Lee J H Heo J H,et al.Cloning and expression anovel esterase gene CPoA from B urkholderia cepacia[J].Journal of Applied Microbiology,2004,96,1306-1316
[27]Saboo V M,Gealt M A1Gene Seguences of.the PCPB gene of Pentachlorophenol-degrading Sphingomonas Chlorophenolica found in nondegrading bacteric[J].Canadian Journal of Microbiology,1998,44 667-675
[28]Praphailong W,M.Van Gestel,Fleet G H,et all Evaluation of the Biolog system for the identification of food and beverage yeasts[J].Letters in Applied Microbiology,1997,24,455-459
[29]Shan Hua Yang,Pi Han Wang.Three species of yeasts news to Taiwan[J].Taiwania,2003,48(2):99-105
[30]张朝华.Biolog细菌自动鉴定系统的应用[J].中国进出境动植检,1994(3):30-32
[31]谢家仪,王永力.BIOLOG细菌自动鉴定系统的应用与研究[J].微生物学通报,1996,23(5):264-267
[32]高崇省,魏亚东,王学令.Biplog细菌自动鉴定系统操作体会[J].中国进出境动植检,1996,(2):26-27
[33]冯瑞华,樊蕙,李力,等.Biolog细菌自动鉴定系统应用初探[J].微生物学杂志,2000,20(2):36-38
[34]李运,盛慧.Biolog微生物鉴定系统在菌种鉴定中的应用[J].酿酒科技,2005,133(7):84-851
[35]虞云龙,樊德方.农药微生物降解的研究现状与发展策略[J].环境科学进展,1996,4(3):28-36
[36]Magdoub MNI,Fayed AE,El-Shenawy MA,Abou-Arab AAK.Persistence of fenvalerate pyrethroid in milk in relation to lactic acid bacteria[J].Egyptian Journal of Dairy Science,1989,17(2):217-225
[37]Misra AK,Vinod RS,Bhatttacharyya A.Degradation of fenvalerate(pyrethroid) pesticide in milk by lactic acid bacteria[J].Indian Journal of Dairy Science,1996,49(9):635-639
[38]Rangaswamy V,Venkateswarlu K.Degradation of selected insecticides monocrotophos,quinalphos,cypermethrin and fenvalerate]by bacteria isolated from soil[J].Bulletin of Environmental Contamination and Toxicology,1992,49(6):797-804
[39]Musumeci MR,Ostiz SB,Binding of cypermethrin residue in Brazilian soils and its release by microbial Activity[J].Revista de Microbiologia,1994,25(4):216-219
[40]Lee S,Gan J,Kim JS,Kabashima JN,Crowley DE.Microbial transformation of pyrethroid insecticides in aqueous and sediment phases[J].Environmental Toxicology and Chemistry,2004,23(1):1-6
[41]Grant RJ,Daniell TJ,Betts WB.Isolation and identification of synthetic pyrethroid degrading bacteria[J].Journal of Applied Microbiology,2002,92(3):534-540
[42]Grant RJ,Betts WB.Biodegradation of the synthetic pyrethroid cypermethrin in used sheep dip.Letters in Applied Microbiology,2003,36(3):173-176
[43]Grant RJ,Betts WB.Mineral and carbon usage of two synthetic pyrethroid degrading bacterial isolates [J].Journal of Applied Microbiology,2004,97(3):656-662
[44]Hashem,Hafez FH,El-Mohandes MAO.Isolation and identification of pyrethroid insecticides degrading bacteria from soil[J].Annals of Agricultural Science,1999,44(1):123-137
[45]戴青华,张瑞福,蒋建东.一株三唑磷降解菌mp-4的分离鉴定及降解特性的研究[J].土壤学报,2005,42(1):111-115
[46]王兆守,梁小虾,林淦.拟除虫菊酯类农药降解菌的紫外线诱变[J].华东昆虫学报,2003,12(2):82-86.
[47]张卫,虞云龙,吴加伦,等.阿维菌素在土壤中的降解和高效降解菌的筛选[J].土壤学报,2004,41(4):590-596
[48]丁海涛,李顺鹏,沈标,崔中利.拟除虫菊酯类农药降解菌的筛选及其生理特性的研究[J].土壤学报,南京:2003,40(1):123-129
[49]育新,戴青华,李晓慧,李顺鹏.氯氰菊酯降解菌株CDT3的分离鉴定及生理特性研究[J].农业环境科学学报,2004,23(5):958-963
[50]Maloney SE.,Maule A,Smith ARW.Purification andpreliminary characterization of permethrinase from a pyrethroid-transforming strain of Bacillus cereus[J].Applied and Environmental Microbiology,1993,59(7):2007-2013
[51]沈德中.污染环境的生物修复(第一版)IM].北京:化学工业出版社,2001:14,311
[52]张从,夏立江.污染土壤生物修复技术[M].北京:中国环境科学出版社,2000:50-65
[53]徐亚同,史家梁,张明.生物修复技术的作用机理和应用[J].上海化工,2001(19):4-7
[54]阎晓明,何金柱.重金属污染土壤的微生物修复机理及研究进展[J].安徽农业科学,2002,30(6):877-879,883
[55]王保军.微生物与重金属的相互作用[J].重庆环境科学,1996,18(1):35-38
[56]Autry A R,Ellis G M.Bioremediation:an effective remedial alternative for petroleum hydrocarbon-contaminatedsoil[J].Environment Progress,1992,11(4):318-323
[57]陈晓东,常文越,邵春岩.土壤污染生物修复技术研究进展[J].环境保护科学,2001,27(5):23-25
[58]张甲耀,李静,夏威林,等.生物修复枝术研究进展[J].应用与环境生物学报,1996,2(2):193-199
[59]Hicks B N,Caplan J A.Bioremediation:A natural solution[J].Pollution Engineering,1993,25(2):30-33
[60]Prince M,Sambasvam Y.Bioremediation of petroleumwastes from the refining of lubricant oils[J].EnvironmentProgress,1993,12(1):5-11
[61]Fredtickson J K,Bolton H,Brockmon F J.Insituandon-situ Bioremediation[J].Evironmental Science Technology,1993,27(9):1711-1761
[62]Dean N.Advancing research for bioremediation[J].Environment Problem,1992,3(7):19-25
[63]Al-Makkawy H.K.,Madbouly M.D.,Persistence and accumulation of some organic insecticides in Nile water and fish.Resources[J].Conservation and Recycling,1999,27(1-2):105-115
[64]Sinha G,Agrawal AK,Islam F,Seth K,Chaturvedi RK,Shukla S,Seth PK.Mosquito repellent (pyrethroid- based) induced dysfunction of blood-brain barrier permeability in developing brain[J].International Journal of Developmental Neuroscience,2004,22(1):31-37
[65]Kolaczinski JH,Curtis CF.Chronic illness as a result of low-level exposure to synthetic pyrethroid insecticides:A review of the debate[J].Food and Chemical Toxicology,2004,42(5):697-706
[66]Mohnssen H M.Chronic sequelae and irreversible injuries following acute pyrethroid intoxication[J].Toxicology Letters,1999,107(1-3):161-176
[67]Shukla Y,Yadav A,Arora A.Carcinogenic and cocar cinogenic potential of cypermethrin on mouse skin [J].Cancer Letters,2002,182(1):33-41
[68]Shukla Y,Arora A,Singh A.Tumourigenic studies on deltamethrin in Swiss albino mice.Toxicology,2001,163(1):1-9
[69]Kasat K,Go V,Pogo BGT.Effects of pyrethroid insecticides and estrogen on WNT10B proto-oncogene expression[J].Environment International,2002,28(5):429-432
[70]Grosman N,Diel F.Influence of pyrethroids and piperonyl butoxide on the Ca2+-ATPase activity of rat brain synaptosomes and leukocyte membranes.International Immunopharmacology,2005,5(2):63-270
[71]Xia YK,Bian Q,Xu LC,Cheng SP,Song L,Liu JI,Wu W,Wang SL,Wang XR.Genotoxic effects on human spermatozoa among pesticide factory workers exposed to fenvalerate[J].Toxicology,2004,203(1-3):49-60
[72]De la Cerda E.Action potential and underlying ionic currents by the pyrethroid insecticide deltamethrin [J].Archives of Medical Research,2002,33(5):448-454
[73]王朝晖,尹伊伟,林小涛,等.拟除虫菊酯农药对水生态系统的生态毒理学研究综述[J].南大学学报(自然科学版),2000,21(3):123-127
[74]Wang LG,Jiang X,Yan DY,Forster S,Martens D.Comparison of two procedures for extraction and clean-up of orgunophosphorus and pyrethroid pesticides in sediment.Pedosphere,2004,14(2):229-234
[75]Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.,Struhl,K.(1995)[M].Short Protocols in Molecular Biology,4th Edition(New York,NK:John Wiley John and Sons Inc.)
[76]Grant R J,Betts W B.Biodegradation of the synthetic pyrethroid eypermethrin inused sheep dip[J].Applied Microbiology.2003(36):173-176
[77]梁定鸿,葛江海.固相萃取气相色谱法测定水中的溴氰菊酯[M].广东,化学工程师.2003,(29-30)
[78]中华人民共和国国家进出口商品检验局.农药残留量气相色谱法[M].北京:中国对外经济贸易出版社,1986,179
[79]默涛,陈鹤鑫,陆贻通,等.农药残留量分析方法[M]上海:科学技术出版社,1992,121-140.
[80]GB/T50091110-2003.植物性食品中氯氰菊酯、氰戊菊酯和溴氰菊酯残留量的测定[S]
[81]Mestres R,Atawijaya S,Cheralier C1 Methods for the study and determination of pestcide in cereal products.Organchlorine.organophosphorus and pyrethroid residues[J].Ann falsif Expert Chem,1979,72:577-589
[82]周良模.气相色谱新技术[M].北京:科学出版社,1994
[83]覃泰群,田淑贤,买光熙.拟除虫菊酯的色谱分析法[J].农业环境护,1994,13(4):192-192
[84]庞国芳,范春林,曹彦忠,等.填充柱气相色谱法同时测定水果、蔬菜和粮谷10种拟除虫菊酯残留量[J].现代商检科技,1995,5(1):1-7
[85]Okadu T,UnoM,Nozawom,Tanigawa.Studies Pyrethroids inseceicide Ⅱ.ECD gas determinatiaon of pyrethroid insecticide in grain[J].Shokulin Eiseigaku Inssli,1983,24,154
[86]王继英,傅承光.羊组织中溴氰菊酯残留量的测定[J].分析测试技术与仪器,1997,3(2):98-108
[87]农业部农药检定所.新编农药手册[M].北京:中国农业出版社,1989
[88]刘均,何燕,蔡克健.溴氰菊酯残留物的分析方法研究[J].云南环境科学,1995,14(2):56-60
[89]Sharp GJ,Brayan J G,Dillis S;Haddad PR,Desmarehelier JM.Extraction,clearup and chromatographic determination of organophosphate,pyrethroid and carabamate grain protectants[J].Analyst,1988,113:1493-1507
[90]《伯杰氏系统细菌学手册》(Bergey's Manual of Systematic Bacteriology,Vol.1-4,1984-1989).
[91]周德庆.微生物学教程[M].北京:高等教育出版社,140-141
[92]俞毓馨,吴国庆.环境工程微生物检测手册[M].中国农业科学出版社,1990.106-110
[2]李凯峰,温青,石汕.污染土壤的生物修复[J].化学工程师,2002,93(6):52-53
[3]杨国栋.污染土壤微生物修复技术主要研究内容和方法[J].农业环境保护,2001,20(4):286-288
[4]Bento F M,Camargo F A,Okeke B C,et al.Comparative bioremediation of soils contaminated with diesel oil by natural attenuation,biostimulation and bioaugmentation[J].Bioresource technology,2005,96:1049-1055
[5]Takeuchi M,Nanba K,Iwamoto,etal.In situbioremediation of a cisdichloroethylene utilizing methanerich groundwater from an uneontaminatedaquifer[J].Water research,2005,2438-2444
[6]滕应,黄昌勇.重金属污染土壤的微生物生态效应及修复研究进展[J].土壤与环境,2002,11(1):85-89
[7]张春桂,许华夏,姜晴楠.污染土壤生物恢复技术[J].生态学杂志,1997,18(4):52-58
[8]Herkovits J,Perez C,Herkovits F D.Ecotoxicological studies of environmental samples from Buenos Airesarea using a standardized amphibian embryo toxicitytest(AMPHITOX)[J].Environmental Pollution,2002,116(1):177-183
[9]霍美玲.葛根素对溴氰菊酯急性中毒的实验治疗[J].卫生毒理学杂志,1988,2(4):124
[10]耿文奎,杨玉英.溴氰菊酯毒性研究现状[J]广西预防医学,1996,2(5):308-310
[11]陈天铎,关育芳.溴氰菊酯对微小牛蜱的敏感度和对雌蜱产卵的影响的研究[J].中国兽医寄生虫病,1994,2(1):6-9
[12]王明学,扶庆,周至刚,等.溴氰菊酯对草鱼早期发育阶段的毒性效应[J].渔业,2000,20(6):39-40
[13]张毓琪,陈叙龙.环境生物毒理学[M].天津:天津大学出版社,1993
[14]贺全仁,聂星湖.拟除虫菊酯类农药的临床毒理学研究现状[J].中国工业医学杂志,1997,10(1):46-48
[15]胡春容.拟除虫菊酯农药的毒性研究进展[J].毒理学杂志,河北,唐山:2005,19(3):239-241
[16]霍美玲.葛根素对溴氰菊酯急性中毒的实验治疗[J].卫生毒理学杂志,1988,2(4):124
[17]Papadopulou,Mourkidou El Analusis ofEstablished Pyrethroids[J].Inseeticidel Residue Rev,1983,89:79-208
[18]刘世亮,骆永明,丁克强,等.土壤中有机污染物的植物修复研究进展[J].土壤,2003,35(3):187-192
[19]董元华,张桃林.基于农产品质量安全的土壤资源管理与可持续利用[J].土壤,2003,35(3):182-186
[20]Liao M,Xie XM,Subhani A.Combined effect of nutrientand pest management on soil ecological quality in hybridrice double-cropping system[J].Pedosphere,2003,13(2):129-138
[21]农业部农药检定所.新编农药手册[M].北京:中国农业出版社,1989
[22]刘均,何燕,蔡克健,溴氰菊酯残留物的分析方法研究[J].云南环境科学,1995,14(2):56-66
[23]Dean N.Advancing research for bioremediation[J].Environment Problem,1992,3(7):19-25
[24]ELROYAC.The Rhizosphere[M].Berlin:Spring-Vedag,1986
[25]Stager C E,Davis J R1 Automated Systems for identification of microorganisms[J].Clinical Microbiology Reviews,1992,5:302-307
[26]Kim C H,Lee J H Heo J H,et al.Cloning and expression anovel esterase gene CPoA from B urkholderia cepacia[J].Journal of Applied Microbiology,2004,96,1306-1316
[27]Saboo V M,Gealt M A1Gene Seguences of.the PCPB gene of Pentachlorophenol-degrading Sphingomonas Chlorophenolica found in nondegrading bacteric[J].Canadian Journal of Microbiology,1998,44 667-675
[28]Praphailong W,M.Van Gestel,Fleet G H,et all Evaluation of the Biolog system for the identification of food and beverage yeasts[J].Letters in Applied Microbiology,1997,24,455-459
[29]Shan Hua Yang,Pi Han Wang.Three species of yeasts news to Taiwan[J].Taiwania,2003,48(2):99-105
[30]张朝华.Biolog细菌自动鉴定系统的应用[J].中国进出境动植检,1994(3):30-32
[31]谢家仪,王永力.BIOLOG细菌自动鉴定系统的应用与研究[J].微生物学通报,1996,23(5):264-267
[32]高崇省,魏亚东,王学令.Biplog细菌自动鉴定系统操作体会[J].中国进出境动植检,1996,(2):26-27
[33]冯瑞华,樊蕙,李力,等.Biolog细菌自动鉴定系统应用初探[J].微生物学杂志,2000,20(2):36-38
[34]李运,盛慧.Biolog微生物鉴定系统在菌种鉴定中的应用[J].酿酒科技,2005,133(7):84-851
[35]虞云龙,樊德方.农药微生物降解的研究现状与发展策略[J].环境科学进展,1996,4(3):28-36
[36]Magdoub MNI,Fayed AE,El-Shenawy MA,Abou-Arab AAK.Persistence of fenvalerate pyrethroid in milk in relation to lactic acid bacteria[J].Egyptian Journal of Dairy Science,1989,17(2):217-225
[37]Misra AK,Vinod RS,Bhatttacharyya A.Degradation of fenvalerate(pyrethroid) pesticide in milk by lactic acid bacteria[J].Indian Journal of Dairy Science,1996,49(9):635-639
[38]Rangaswamy V,Venkateswarlu K.Degradation of selected insecticides monocrotophos,quinalphos,cypermethrin and fenvalerate]by bacteria isolated from soil[J].Bulletin of Environmental Contamination and Toxicology,1992,49(6):797-804
[39]Musumeci MR,Ostiz SB,Binding of cypermethrin residue in Brazilian soils and its release by microbial Activity[J].Revista de Microbiologia,1994,25(4):216-219
[40]Lee S,Gan J,Kim JS,Kabashima JN,Crowley DE.Microbial transformation of pyrethroid insecticides in aqueous and sediment phases[J].Environmental Toxicology and Chemistry,2004,23(1):1-6
[41]Grant RJ,Daniell TJ,Betts WB.Isolation and identification of synthetic pyrethroid degrading bacteria[J].Journal of Applied Microbiology,2002,92(3):534-540
[42]Grant RJ,Betts WB.Biodegradation of the synthetic pyrethroid cypermethrin in used sheep dip.Letters in Applied Microbiology,2003,36(3):173-176
[43]Grant RJ,Betts WB.Mineral and carbon usage of two synthetic pyrethroid degrading bacterial isolates [J].Journal of Applied Microbiology,2004,97(3):656-662
[44]Hashem,Hafez FH,El-Mohandes MAO.Isolation and identification of pyrethroid insecticides degrading bacteria from soil[J].Annals of Agricultural Science,1999,44(1):123-137
[45]戴青华,张瑞福,蒋建东.一株三唑磷降解菌mp-4的分离鉴定及降解特性的研究[J].土壤学报,2005,42(1):111-115
[46]王兆守,梁小虾,林淦.拟除虫菊酯类农药降解菌的紫外线诱变[J].华东昆虫学报,2003,12(2):82-86.
[47]张卫,虞云龙,吴加伦,等.阿维菌素在土壤中的降解和高效降解菌的筛选[J].土壤学报,2004,41(4):590-596
[48]丁海涛,李顺鹏,沈标,崔中利.拟除虫菊酯类农药降解菌的筛选及其生理特性的研究[J].土壤学报,南京:2003,40(1):123-129
[49]育新,戴青华,李晓慧,李顺鹏.氯氰菊酯降解菌株CDT3的分离鉴定及生理特性研究[J].农业环境科学学报,2004,23(5):958-963
[50]Maloney SE.,Maule A,Smith ARW.Purification andpreliminary characterization of permethrinase from a pyrethroid-transforming strain of Bacillus cereus[J].Applied and Environmental Microbiology,1993,59(7):2007-2013
[51]沈德中.污染环境的生物修复(第一版)IM].北京:化学工业出版社,2001:14,311
[52]张从,夏立江.污染土壤生物修复技术[M].北京:中国环境科学出版社,2000:50-65
[53]徐亚同,史家梁,张明.生物修复技术的作用机理和应用[J].上海化工,2001(19):4-7
[54]阎晓明,何金柱.重金属污染土壤的微生物修复机理及研究进展[J].安徽农业科学,2002,30(6):877-879,883
[55]王保军.微生物与重金属的相互作用[J].重庆环境科学,1996,18(1):35-38
[56]Autry A R,Ellis G M.Bioremediation:an effective remedial alternative for petroleum hydrocarbon-contaminatedsoil[J].Environment Progress,1992,11(4):318-323
[57]陈晓东,常文越,邵春岩.土壤污染生物修复技术研究进展[J].环境保护科学,2001,27(5):23-25
[58]张甲耀,李静,夏威林,等.生物修复枝术研究进展[J].应用与环境生物学报,1996,2(2):193-199
[59]Hicks B N,Caplan J A.Bioremediation:A natural solution[J].Pollution Engineering,1993,25(2):30-33
[60]Prince M,Sambasvam Y.Bioremediation of petroleumwastes from the refining of lubricant oils[J].EnvironmentProgress,1993,12(1):5-11
[61]Fredtickson J K,Bolton H,Brockmon F J.Insituandon-situ Bioremediation[J].Evironmental Science Technology,1993,27(9):1711-1761
[62]Dean N.Advancing research for bioremediation[J].Environment Problem,1992,3(7):19-25
[63]Al-Makkawy H.K.,Madbouly M.D.,Persistence and accumulation of some organic insecticides in Nile water and fish.Resources[J].Conservation and Recycling,1999,27(1-2):105-115
[64]Sinha G,Agrawal AK,Islam F,Seth K,Chaturvedi RK,Shukla S,Seth PK.Mosquito repellent (pyrethroid- based) induced dysfunction of blood-brain barrier permeability in developing brain[J].International Journal of Developmental Neuroscience,2004,22(1):31-37
[65]Kolaczinski JH,Curtis CF.Chronic illness as a result of low-level exposure to synthetic pyrethroid insecticides:A review of the debate[J].Food and Chemical Toxicology,2004,42(5):697-706
[66]Mohnssen H M.Chronic sequelae and irreversible injuries following acute pyrethroid intoxication[J].Toxicology Letters,1999,107(1-3):161-176
[67]Shukla Y,Yadav A,Arora A.Carcinogenic and cocar cinogenic potential of cypermethrin on mouse skin [J].Cancer Letters,2002,182(1):33-41
[68]Shukla Y,Arora A,Singh A.Tumourigenic studies on deltamethrin in Swiss albino mice.Toxicology,2001,163(1):1-9
[69]Kasat K,Go V,Pogo BGT.Effects of pyrethroid insecticides and estrogen on WNT10B proto-oncogene expression[J].Environment International,2002,28(5):429-432
[70]Grosman N,Diel F.Influence of pyrethroids and piperonyl butoxide on the Ca2+-ATPase activity of rat brain synaptosomes and leukocyte membranes.International Immunopharmacology,2005,5(2):63-270
[71]Xia YK,Bian Q,Xu LC,Cheng SP,Song L,Liu JI,Wu W,Wang SL,Wang XR.Genotoxic effects on human spermatozoa among pesticide factory workers exposed to fenvalerate[J].Toxicology,2004,203(1-3):49-60
[72]De la Cerda E.Action potential and underlying ionic currents by the pyrethroid insecticide deltamethrin [J].Archives of Medical Research,2002,33(5):448-454
[73]王朝晖,尹伊伟,林小涛,等.拟除虫菊酯农药对水生态系统的生态毒理学研究综述[J].南大学学报(自然科学版),2000,21(3):123-127
[74]Wang LG,Jiang X,Yan DY,Forster S,Martens D.Comparison of two procedures for extraction and clean-up of orgunophosphorus and pyrethroid pesticides in sediment.Pedosphere,2004,14(2):229-234
[75]Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.,Struhl,K.(1995)[M].Short Protocols in Molecular Biology,4th Edition(New York,NK:John Wiley John and Sons Inc.)
[76]Grant R J,Betts W B.Biodegradation of the synthetic pyrethroid eypermethrin inused sheep dip[J].Applied Microbiology.2003(36):173-176
[77]梁定鸿,葛江海.固相萃取气相色谱法测定水中的溴氰菊酯[M].广东,化学工程师.2003,(29-30)
[78]中华人民共和国国家进出口商品检验局.农药残留量气相色谱法[M].北京:中国对外经济贸易出版社,1986,179
[79]默涛,陈鹤鑫,陆贻通,等.农药残留量分析方法[M]上海:科学技术出版社,1992,121-140.
[80]GB/T50091110-2003.植物性食品中氯氰菊酯、氰戊菊酯和溴氰菊酯残留量的测定[S]
[81]Mestres R,Atawijaya S,Cheralier C1 Methods for the study and determination of pestcide in cereal products.Organchlorine.organophosphorus and pyrethroid residues[J].Ann falsif Expert Chem,1979,72:577-589
[82]周良模.气相色谱新技术[M].北京:科学出版社,1994
[83]覃泰群,田淑贤,买光熙.拟除虫菊酯的色谱分析法[J].农业环境护,1994,13(4):192-192
[84]庞国芳,范春林,曹彦忠,等.填充柱气相色谱法同时测定水果、蔬菜和粮谷10种拟除虫菊酯残留量[J].现代商检科技,1995,5(1):1-7
[85]Okadu T,UnoM,Nozawom,Tanigawa.Studies Pyrethroids inseceicide Ⅱ.ECD gas determinatiaon of pyrethroid insecticide in grain[J].Shokulin Eiseigaku Inssli,1983,24,154
[86]王继英,傅承光.羊组织中溴氰菊酯残留量的测定[J].分析测试技术与仪器,1997,3(2):98-108
[87]农业部农药检定所.新编农药手册[M].北京:中国农业出版社,1989
[88]刘均,何燕,蔡克健.溴氰菊酯残留物的分析方法研究[J].云南环境科学,1995,14(2):56-60
[89]Sharp GJ,Brayan J G,Dillis S;Haddad PR,Desmarehelier JM.Extraction,clearup and chromatographic determination of organophosphate,pyrethroid and carabamate grain protectants[J].Analyst,1988,113:1493-1507
[90]《伯杰氏系统细菌学手册》(Bergey's Manual of Systematic Bacteriology,Vol.1-4,1984-1989).
[91]周德庆.微生物学教程[M].北京:高等教育出版社,140-141
[92]俞毓馨,吴国庆.环境工程微生物检测手册[M].中国农业科学出版社,1990.106-110