三株轻度嗜盐反硝化菌的分离鉴定和降解特性初探
|
摘要
近年来由于工农业的快速发展,排放到水体中的氮素逐年增多,造成严重的水体富营养化,引起水质恶化、湖泊退化,也危害到人体的健康。
生物脱氮工艺因具有处理效果好、处理成本低,操作管理方便等优点而得到广泛应用。在石油、化工、海产品加工和养殖等行业中,外排含氮废水大多具有高盐的特点。研究表明,高盐环境下的盐析作用致使微生物的脱氢酶活性降低、导致微生物细胞破裂而死亡,可见高盐浓度对生物脱氮带来不利影响。因此高盐这一特殊生境中反硝化细菌的分离培养和降解特性研究将有助于了解高盐生物脱氮工艺的可行性。
本论文在实验室已驯化成熟的活性污泥基础上,采用硝酸钾作为惟一氮源,进行嗜盐反硝化菌的富集、分离和筛选,得到六株具有反硝化性能的嗜盐菌株,根据各菌株的脱氮效果优选降解性能较好的三株菌进行后续试验。
通过对三株菌的生理生化试验、16S rDNA测序和同源性分析可得:菌株FX1属于假单胞菌(Pseudomonas sp.),菌株YL-1属于迪茨氏菌(Dietzia sp.),菌株GYL属于盐单胞菌(Halomonas sp.)。
考察了盐度、DO、接种量、pH、温度、碳源种类和C/N对三株菌降解性能的影响,得到各菌株的最佳降解条件。
研究发现,三株菌在异养硝化培养基中均具有较好的异养硝化效果,在改进的DM培养基中异养硝化效果更好。探讨了菌株FX1的好氧反硝化途径与异养硝化途径相互间的影响,发现该菌株的好氧反硝化作用可以促进异养硝化反应的进行。
三株菌既具有异养硝化作用,又具有好氧反硝化作用,可见三株菌可实现同步硝化反硝化(SND),即可以独立完成生物脱氮的全过程。由于三株菌均可耐受一定的盐度,并能实现同步硝化反硝化,因此三株菌对于高盐度废水的处理具有很高的应用价值。
In recent years,due to the rapid development of industry and agriculture,the content of nitrate in the wastewater discharged into the water is becoming higher and higher and have caused serious eutrophication,the quality of water depravation and the lake degenerate.Sometimes,the contaminated water will threaten the people health as the water source.
The biological nitrogen removal process has many advantages such as cheap in cost, easy in operation,convenient in treatment so it has been widely used in wastewater treatment.In petroleum,chemical,seafood process industry,most of the discharge is hypersaline wastewater.In high-salt environment,the salting-out effect may decrease the microorganisms' dehydrogenase activity,lead to microbial cells rupture.Sometimes,the salting-out effect even cause death of microbial.So the high salt concentration has a negative impact on the biological nitrogen removal effect.Therefore,the isolation of denitrifying bacteria from hyperhaline habitats will provide a reference value for hyperhaline biological nitrogen removal process.
In this paper,we have accomplished the enrichment,islation and screening experiments from the salt-tolerant sludge systems using nitrate as the sole nitrogen source. Six strains of halotolerant denitrifying bacteria were preliminary islated from the activated sludge.According to the degradation characteristics of various strains,we selected three of them for the following tests.
According to the morphological observation,physiological biochemical test and sequence analysis of the 16S rDNA,strain FX1 was identified as Pseudomonas sp.;strain YL-1 was identified as Dietzia sp.;strain GYL was identified as Halomonas sp..
We have investigated seven different effect factors effecting the degradation characteristics such as salinity,DO,inoculum concentration and so on to get the optimal degradation condition of each strain.
The heterotrophic nitrification perfermance of the three bacteria has been found. When these three bacteria were cultured in the DM medium,they have higher nitrogen removal efficiency.And the relationship of aerobic denitrification and heterotrophic nitrification perfermance about FX1 was discussed.The result showed that aerobic denitrification can promote the effect ofheterotrophic nitrification.
These three bacteria are aerobic denitrifying bacteria which can simultaneously perform heterotrophic nitrification,so they can realize simultaneous nitrification and denitrification(SND).
The three bacteria can achieve SND in hyperhaline environment,so our tests have big application value for nitrogen removal of hyperhaline wastewater.
生物脱氮工艺因具有处理效果好、处理成本低,操作管理方便等优点而得到广泛应用。在石油、化工、海产品加工和养殖等行业中,外排含氮废水大多具有高盐的特点。研究表明,高盐环境下的盐析作用致使微生物的脱氢酶活性降低、导致微生物细胞破裂而死亡,可见高盐浓度对生物脱氮带来不利影响。因此高盐这一特殊生境中反硝化细菌的分离培养和降解特性研究将有助于了解高盐生物脱氮工艺的可行性。
本论文在实验室已驯化成熟的活性污泥基础上,采用硝酸钾作为惟一氮源,进行嗜盐反硝化菌的富集、分离和筛选,得到六株具有反硝化性能的嗜盐菌株,根据各菌株的脱氮效果优选降解性能较好的三株菌进行后续试验。
通过对三株菌的生理生化试验、16S rDNA测序和同源性分析可得:菌株FX1属于假单胞菌(Pseudomonas sp.),菌株YL-1属于迪茨氏菌(Dietzia sp.),菌株GYL属于盐单胞菌(Halomonas sp.)。
考察了盐度、DO、接种量、pH、温度、碳源种类和C/N对三株菌降解性能的影响,得到各菌株的最佳降解条件。
研究发现,三株菌在异养硝化培养基中均具有较好的异养硝化效果,在改进的DM培养基中异养硝化效果更好。探讨了菌株FX1的好氧反硝化途径与异养硝化途径相互间的影响,发现该菌株的好氧反硝化作用可以促进异养硝化反应的进行。
三株菌既具有异养硝化作用,又具有好氧反硝化作用,可见三株菌可实现同步硝化反硝化(SND),即可以独立完成生物脱氮的全过程。由于三株菌均可耐受一定的盐度,并能实现同步硝化反硝化,因此三株菌对于高盐度废水的处理具有很高的应用价值。
In recent years,due to the rapid development of industry and agriculture,the content of nitrate in the wastewater discharged into the water is becoming higher and higher and have caused serious eutrophication,the quality of water depravation and the lake degenerate.Sometimes,the contaminated water will threaten the people health as the water source.
The biological nitrogen removal process has many advantages such as cheap in cost, easy in operation,convenient in treatment so it has been widely used in wastewater treatment.In petroleum,chemical,seafood process industry,most of the discharge is hypersaline wastewater.In high-salt environment,the salting-out effect may decrease the microorganisms' dehydrogenase activity,lead to microbial cells rupture.Sometimes,the salting-out effect even cause death of microbial.So the high salt concentration has a negative impact on the biological nitrogen removal effect.Therefore,the isolation of denitrifying bacteria from hyperhaline habitats will provide a reference value for hyperhaline biological nitrogen removal process.
In this paper,we have accomplished the enrichment,islation and screening experiments from the salt-tolerant sludge systems using nitrate as the sole nitrogen source. Six strains of halotolerant denitrifying bacteria were preliminary islated from the activated sludge.According to the degradation characteristics of various strains,we selected three of them for the following tests.
According to the morphological observation,physiological biochemical test and sequence analysis of the 16S rDNA,strain FX1 was identified as Pseudomonas sp.;strain YL-1 was identified as Dietzia sp.;strain GYL was identified as Halomonas sp..
We have investigated seven different effect factors effecting the degradation characteristics such as salinity,DO,inoculum concentration and so on to get the optimal degradation condition of each strain.
The heterotrophic nitrification perfermance of the three bacteria has been found. When these three bacteria were cultured in the DM medium,they have higher nitrogen removal efficiency.And the relationship of aerobic denitrification and heterotrophic nitrification perfermance about FX1 was discussed.The result showed that aerobic denitrification can promote the effect ofheterotrophic nitrification.
These three bacteria are aerobic denitrifying bacteria which can simultaneously perform heterotrophic nitrification,so they can realize simultaneous nitrification and denitrification(SND).
The three bacteria can achieve SND in hyperhaline environment,so our tests have big application value for nitrogen removal of hyperhaline wastewater.
引文
[1]朱济成.关于地下水硝酸盐污染原因的探讨[J].北京地质,1995,(2):20-26
[2]李庆诚.全国重点环境考核城市地下水污染概况及防治对策[J].地下水,1992,14(1):2-5
[3]白娟,郑姝卉,吴秋兰.水资源现状探讨[J].现代农业科技,2007(12):187-188
[4]中国工程院“21世纪中国可持续发展水资源战略研究”项目组.中国可持续发展水资源战略研究综合报告[J].中国工程科学,2000,2(8):1-17
[5]祝兴祥等.中国氮、磷污染现状及控制标准,污水除磷脱氮技术研究与实践,中国土木工程学会水工业分会排水委员会.2000
[6]2007年《中国环境状况公报》2008年6月4日公布
[7]郑兴灿,李亚新,污水除磷脱氮技术.北京:中国建筑工业出版社,1998:7-9
[8]叶剑峰等.废水生物脱氮处理新技术.北京:化学工业出版社,2006:12-13
[9]周奇迹.农业微生物学.北京:中国农业出版社,2001:125-126
[10]L.A.Robertson,J.G.Kuenen.Aerobic Denitrification:a Controversy Revived[J].Arch.Microbiol,1984,139(5):351-354
[11]Takaya N,Maria Antonina B C S,Yasushi S,et al.Aerobic denitrification bacteria that produce low levels of nitrous oxide[J].App and Envir Microbio,2003,69(6):3152-3157
[12]Wilson L P,Bouwer E J.Biodegradation of aromatic compounds under mixed oxygen /denitrifying conditions[J].Industr Microbiol Biotech,1997,18:116-130
[13]Bell LC,Richardson DJ,Ferguson SJ.Periplasmic and membrane-bound nitrate reductase in Thiosphaera pantothropa[J].FEBS Left,1990,265:85-87
[14]Bell LC,Berks BC,Richardson DJ,et al.Insertion of transportation Tn5 into a structural gene of themembrane-bound nitrate reudctase of Thiosphaera pantothropa results in anaerobic over expression of Periplasmic nitrate reductase activity[J].Gen Microbial,1993,139:3205-3214
[15]Farver O,Kroneck P M H,Zumft W G,et al.Allosteric control of internal electron transfer in cytochrome cd1 nitrite reductase[J].PNAS,2003,100:7622-7625
[16]Zajicek R S,Allen J W,Richardson D J,et al.Paracoccus pan-totrophus NapC can reductively activate cytochrome cd1 nitrite reductase[J].FEBS LETT,2004,565:48-52
[17]Ye R.W,Averill B.A,Tiedje J.M.Denitrification:Production and consumption of nitricoxide[J].Environ Microbiot,1994,60:281-286
[18]Carr GJ,Page M.D,Fergtlson S.J,et al.The energy-conserving nitric-oxide-reductase system in Paracoccus denitrifican Distrbution,from the nitrite reductase that catalyses synthesis of nitric oxide and evldence from trapping experiments for nitric oxide as a free intermediate during denitrification[J].Eur.J.Biochem,1989,179:686-692
[19]Car G.J,Feruson S.J.The nitric oxide reductase of Paracoccus denitrificans[J].Biochem,1990,269:423-430
[20]Heiss B,Frunzke K,W Zumft.Formation of N=N bond from nitric oxide by a membrane-bound cytochrome bc complex of Nitrate-respiring(denilrifying)Psedomonoas stuteri[J].Bacteriol,1989,171:3288-3297
[21]Heikkila M P,Honisch U,Wunsch P,et al.Role of the tat transport system in nitrous oxide reductase translocation and cytochrome cd1 biosynthesus in Psudomonas stutzeri[J].Journal of Bacteriology,2001,183:1663-1671
[22]殷芳芳,王淑莹,昂雪野等.碳源类型对低温条件下生物反硝化的影响[J].环境科学,2009,30(1):108-113
[23]于爱茸,李尤,俞吉安.一株耐氧反硝化菌的筛选及脱氮特性研究[J].微生物学杂,2005,25(3):77-81
[24]Huang H.K,Tseng SK.Nitrate reduction by a citrobacter diversus under aerobic environment[J].Appl Microbiol Biotechno],2001,55:90-94
[25]殷士学,陈丽敏.土壤中硝化、反硝化微生物的研究进展[J].土壤学报,2000,39:116-125
[26]杨基先,高珊珊,马放等.一株好氧反硝化细菌的分离鉴定及反硝化能力[J].环境科学报,2008,28(7):1301-1307
[27]Katarzyna Bernat,Irena Wojnowska-Baryla.Carbon source in aerobic denitrification[J].Biochemical Engineering Journal,2007,36:l16 -122
[28]曾庆武,梁运祥,葛向阳.反硝化细菌的分离筛选及其反硝化特性的初步研究[J].华中农业大学学报,2008,27(5):617-620
[29]Charles GIass,Joann silver.Denitrification kinetics of high nitrate concentration water:pH effecton inhibition and nitrite accumulation[J].Wat.Res,1998,32(3):831-839
[30]Liu L.H,Koenig A.Use of limestone for pH control in autotrophic denitrification:batchexperiments[J].Process Biochemistry,2002,37:885-893
[31]谢珊,李小明,曾光明等.同步硝化反硝化实现途径的探讨[J].环境科学与技术,2004,27(2):107-109.
[32]Alexander M.Introduction to soil microbiology.John Wiley and Sons,NewYork,1997
[33]李树刚,马光庭.生物脱氮菌种筛选及条件选择的研究[J].广西大学学报(自然科学版),2003,28(2):173-176
[34]李卿.好氧反硝化细菌的筛选和生理特性的研究:[硕士学位论文]长春:吉林大学,2006.
[35]阮立权,陈坚.好氧颗粒污泥同步硝化反硝化脱氮过程中N_2O的产生[J].无锡轻工业大学学报,2004,23(4):37-64
[36]Sunjin Hwang,Keisuke Hanaki.Effects of oxygen concentration and moisture content of refuse on nitrification,denitrification and nitrous oxide Production[J].Bioresource Technology,2000,71:159-165
[37]徐亚同.废水生物处理的运行和管理.上海:华东师范大学出版社,1989:181页
[38]Su J J,Liu B Y,Liu C Y.Comparison of aerobic denitrification under high oxygen at mosphereby Thiosphaera pantotropha ATCC35512 and Pseudomonas stutzeri SU2 newly isolated from the actived sludge of a piggery wastewater treatment system[J].Journal of Applied Microbio,2001,90:457-462
[39]Huang H K,Tseng S K.Nitrate reduction by Citrobacter diversus under aerobic environment[J].Appl Microbiol Biotechnol,2001,55:90-94
[40]Patureau D,Zumstein E,Delgenes J P,et al.Aerobic denitrification isolation from diverse natural and managed ecosystems[J].Microbiol Ecol,2000,39:145-152
[41]Lukow T,Diekmann H.Aerobic denitrification by a newly heterotrophic bacterium strain TL1[J].Biotechnol Lett,1997,11(19):1157-1159
[42]Chen F,Xia Q,Ju L K.Aerobic denitrification of Pseudomonas aeruginosa monitored by online NAD(P)H fluorescence[J].Appl and Environ Microbiol,2003,69(11):6715-6722
[43]Kesserli P,Kiss I,Bihari Z,et al.Biological denitrification in a continuous-flow pilot bioreactor containing immobilized Pseudomonas butannovora cells[J].Bioreso Technol,2003,87:75-80
[44]Kim Y J,Yoshizawa M,Takenaka S,et al.Isolation and culture conditions of a Klebsiella pneumoniae strain that can utilize ammonium and nitrate ions simultaneously with controlled iron and molybdate ion concentrations[J].Biosci Biotechnol Biochem,2002,66(5):996-1001
[45]郑平.环境微生物学.浙江:浙江大学出版社,2003
[46]Brierley EDR,Wood M.Heterotrophic nitrification in an acid forest soil:isolation and characterisation of a nitrifying bacterium[J].Soil Biology & Biochemistry,2001,33(10):1403-1409
[47]Schmidt I,Sliekers O,Schmid M,et al.New concepts of microbial treatment processes for the nitrogen removal in wastewater[J].FEMS Microbiology Reviews,2003,27(4):481-492
[48]杨宗政,王鑫,庞金钊等.异养硝化菌的分离及其强化活性污泥脱氮效果[J].中国给水排水, 2006,22(21):67-70
[49]张彤,庞金钊,杨宗政等.异养硝化菌的脱氮研究[J].水科学与工程技术,2006,6:41-42
[50]苟莎,黄钧.异养硝化细菌脱氮特性及研究进展[J].微生物学通报,2009,36(2):255-260
[51]Gupta AB.Thiosphaera pantotropha:a sulphur bacterium capable of simultaneoush eterotrophic nitrification and aerobic denitrification[J].Enzyme and Microbial Technology,1997,21:589-595
[52]何霞,吕剑,何义亮.异养硝化机理的研究进展[J].微生物学报,2006,46(5):844-847
[53]Wehrfritz JM,Carter JP,Spiro S,et al.Hydroxylamine oxidation in heterotrophic nitrate-reducing soil bacteria and purification of a hydroxylamine-cytochrome oxidoreductase from a Pseudomonas species[J].Arch Microbiol,1997,166:421-424
[54]Joo HS,Hirai M,Shoda M.Nitrification and denitrification in high-strength ammonium by Alcaligenes faecalis[J].Biotechnology Letters,2005,27(11):773-778
[55]Hu TL,Kung KT.Study of heterotrophic nitrifying bacteria from wastewater treatment systems treating acrylonitrile,butadiene and styrene resin wastewater[J].Water Science and Technology 2000,42(3-4):315-321
[56]陈赵芳,尹立红,浦跃朴等.一株异养硝化菌的筛选及其脱氮条件[J].东南大学学报(自然科学版),2007,37(3):486-490
[57]Su JJ,Yeh KS,Tsheng PW.A strain of Pseudomonas sp.isolated from piggery wastewater treatment systems with heterotrophic nitrification capability in Taiwan[J].Current Microbiology,2006,53(1):77-81
[58]Kim JK,Park KJ,Cho KS,et al.Aerobic nitrification-denitrification by heterotrophic Bacillus strains[J].Bioresource Technology,2005,96(17):1897-1906
[59]何霞,赵彬,吕剑等.异养硝化细菌Bacillus sp.LY脱氮性能研究[J].环境科学,2007,28(6):1404-1408
[60]Blagodatsky SA,Kesik M,Papen H,et al.Production of NOand N_2O by the heterotrophic nitrifier Alcaligenes faecalis subsp,parafaecalis under varying conditions of oxygen saturation[J].Geomicrobiology Journal,2006,23(3-4):165-176
[61]曹喜涛,常志州,黄红英等.1株高温异养硝化细菌的分离鉴定和特性研究[J].安徽农业科学2006,34(19):4833-4834
[62]林燕,何义亮,李春杰等.MBR同步硝化反硝化及异养硝化试验研究[J].环境科学与技术,2006,29(1):7-9
[63]Anshuman A,Khardenavis,Atya Kapley,et al.Simultaneous nitrification and denitrification by diverse Diaphorobacter sp[J].Appl Microbiol Biotechnol,2007,77:403-409
[64]林燕,孔海南,何义亮等.异养硝化细菌的分离及其硝化特性实验研究[J].环境科学,27(2):324-328
[65]Hung-Soo Joo,Mitsuyo Hirai,Makoto Shoda.Characteristics of Ammonium Removal by Heterotrophic Nitrification-Aerobic Denitrification by Alcaligenes faecalis No.4[J].Journal of Bioscience and Bioengineering.2005,100(2):184-191
[66]刘正.高浓度含盐废水生物处理技术[J].化工环保,2004,24(增刊):209-211
[67]东秀珠,蔡妙英等.常见细菌系统鉴定手册.北京:科学出版社,2001:62-65
[68]R.E布坎南,N.E吉本斯等编.伯杰细菌鉴定手册(第八版).科学出版社,1984
[69]王弘宇,马放,苏俊峰等.好氧反硝化菌株的鉴定及其反硝化特性研究[J].环境科学.2007,28(7):1548-1552
[70]孔庆鑫.一株新型的脱氮微生物的分离鉴定及其脱氮机制.城市环境与城市生态[J].2005,13(2):36-39
[71]崔家荣.水中氨氮纳氏试剂分光光度法测定[J].现代农业科技,2008,8:208-209
[72]国家环境保护总局和《水和废水监测分析方法》编委会编.水和废水监测分析方法.北京:中国环境科学出版社,271-274
[73]孙仕萍,邢大荣,张岚等.水中硝酸盐氮的麝香草酚分光光度测定法[J].环境与健康杂志,2007,24(4):256-257
[74]Chun-ChinWang,Chi-Mei Lee.Isolation of the caprolactam denitrifying bacteria from a Wastewater treatment system manufactured with acrylonitrile-butadiene-styrene resin[J].Journal of Hazardous Materials 2007,145:136-141
[75]苏俊峰,王继华,马放等.好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究[J].环境科学2007,28(10):2332-2335
[76]HAMID-REZA KARIMINIAAE-HAMEDAANI,KOHZO KANDA,FUMIO KATO.Denitrification Activity of the Bacterium Pseudomonas sp.ASM-2-3 Isolated from the Ariake Sea Tideland[J].Journal of Bioscience and Bioengineering,2004,97(1),39-44
[77]项慕飞.好氧反硝化菌的分离筛选和鉴别研究:[硕士学位论文]北京:北京工商大学,2007
[78]刘水平,罗志勇.琼脂糖凝胶电泳实验技巧[J].实用预防医学,2006,13(4):1068-1069
[79]Kesser U P,Kiss I,Bihari Z,et al.Biological denitrificationin a continuous 2 flow pilot bioreactor containing immobilized Pseudomonas butnnovora cells[J].Bioresource Technology,2003,87:75-80
[80]Takaya N,Maria A ntoninaB C S,Yasushi S,et al.Aerobic denitrification bacteria that produce low levels of nitrous oxide[J].App and Envir Microbio,2003,69(6):3152-3157.
[82]燕永亮,杨剑,陈立宏等.斯氏假单胞菌(Pseudomonas stutzeri)A1501反硝化相关基因结构 及功能分析[J].中国科学C辑,2005,35(3):246-253
[83]周丽华,陈士超,邓志瑞等.太湖沉积物中的可培养细菌:Ⅰ.细菌多样性初步分析[J].湖泊科学,2009,21(1):27-35
[84]Irene vonder Weid,Joana M.Marques,Claudia D.Cunha,et al.Identification and biodegradation potential of a novel strain of Dietzia cinnamea isolated from a petroleum-contaminated tropical soil[J].Systematic and Applied Microbiology,2007,30:331-339
[85]刘真,邵宗泽.南海深海沉积物烷烃降解菌的富集分离与多样性初步分析[J].微生物学报,2007,47(5):869-873
[86]A.A.Shapovalova,T.V.Khijniak,T.P.Tourov,et al.Heterotrophic denitrification at extremely high salt and pH by haloalkaliphilic Gammaproteobacteria from hypersaline soda lakes[J].Extremophiles,2008,12:619-625
[87]Sachiko Yoshie,Hiroshi Makino,Hidenobu Hirosawa et al.Molecular analysis of halophilic bacterial community for high-rate denitrification of saline industrial wastewater[J].Appl Microbiol Biotechnol,2006,72:182-189
[88]宁卓,张波.嗜盐菌的研究进展及应用[J].苏盐科技,2007,3(1):31-32
[89]王振雄,徐毅,周培瑾.嗜盐碱古生菌新种的系统分类学研究[J].微生物学报,2000,40(2):115-120
[90]马放,周丹丹,王宏宇等.一株好氧反硝化细菌生理生态特性的研究[J].哈尔滨工业大学学报,2006,38(4):575-577
[91]P.Elefsiniotis,D.Li.The effect of temperature and carbon source on denitrification using volatile fatty acids[J].Biochemical Engineering Journal,2006,28:148-155
[92]杨莎莎,宋英豪,赵宗升等.pH值和碳氮比对亚硝酸型反硝化影响的研究[J].环境工程学报,2007,12(1):15-19
[93]闫博.高效反硝化菌的分离及其在水体中脱氮规律的研究:[硕士学位论文]天津:天津大学,2002
[94]赵庆.破源浓度对反硝化聚磷效果的影响研究:[硕士学位论文]西安:西安建筑科技大学,2003
[95]Hung-Soo Joo,Mitsuyo Hirai,Makoto Shoda.Piggery wastewater treatment using Alcaligenes faecalis strain No.4 with heterotrophic nitrification and aerobic denitrification[J].Water Research,2006,(40):3029-3036
[96]Reto Manser,Willi Gujer,Hansruedi Siegrist.Oecay processes of nitrifying bacteria in biological wastewater treatment systems.Water Research,2006,(40):2416-2426
[97]苏俊峰.异养型同步硝化反硝化脱氮技术及微生物特性研究:[工学博士学位论文].哈尔滨: 哈尔滨工业大学,2007
[98]Soren Rysgaard,Peter Thastu M,Tage Dalsgaard.Effects of Salinityon NH_4~+-adsorption capacity,Nitrification,and Denitrification in Danish Estuarine Sediments.Estuaries,1999,22(3):21-30
[99]柯国华,汪苹,杨志.好氧反硝化脱氮气态中间产物的研究分析[J].环境科学与技术,2006,29(7):45-54
[100]汪苹,王斌,刘军.同步硝化反硝化处理氨氮废水过程中气态脱氮产物的研究[J].环境科学研究,2005,18(5):67-71
[2]李庆诚.全国重点环境考核城市地下水污染概况及防治对策[J].地下水,1992,14(1):2-5
[3]白娟,郑姝卉,吴秋兰.水资源现状探讨[J].现代农业科技,2007(12):187-188
[4]中国工程院“21世纪中国可持续发展水资源战略研究”项目组.中国可持续发展水资源战略研究综合报告[J].中国工程科学,2000,2(8):1-17
[5]祝兴祥等.中国氮、磷污染现状及控制标准,污水除磷脱氮技术研究与实践,中国土木工程学会水工业分会排水委员会.2000
[6]2007年《中国环境状况公报》2008年6月4日公布
[7]郑兴灿,李亚新,污水除磷脱氮技术.北京:中国建筑工业出版社,1998:7-9
[8]叶剑峰等.废水生物脱氮处理新技术.北京:化学工业出版社,2006:12-13
[9]周奇迹.农业微生物学.北京:中国农业出版社,2001:125-126
[10]L.A.Robertson,J.G.Kuenen.Aerobic Denitrification:a Controversy Revived[J].Arch.Microbiol,1984,139(5):351-354
[11]Takaya N,Maria Antonina B C S,Yasushi S,et al.Aerobic denitrification bacteria that produce low levels of nitrous oxide[J].App and Envir Microbio,2003,69(6):3152-3157
[12]Wilson L P,Bouwer E J.Biodegradation of aromatic compounds under mixed oxygen /denitrifying conditions[J].Industr Microbiol Biotech,1997,18:116-130
[13]Bell LC,Richardson DJ,Ferguson SJ.Periplasmic and membrane-bound nitrate reductase in Thiosphaera pantothropa[J].FEBS Left,1990,265:85-87
[14]Bell LC,Berks BC,Richardson DJ,et al.Insertion of transportation Tn5 into a structural gene of themembrane-bound nitrate reudctase of Thiosphaera pantothropa results in anaerobic over expression of Periplasmic nitrate reductase activity[J].Gen Microbial,1993,139:3205-3214
[15]Farver O,Kroneck P M H,Zumft W G,et al.Allosteric control of internal electron transfer in cytochrome cd1 nitrite reductase[J].PNAS,2003,100:7622-7625
[16]Zajicek R S,Allen J W,Richardson D J,et al.Paracoccus pan-totrophus NapC can reductively activate cytochrome cd1 nitrite reductase[J].FEBS LETT,2004,565:48-52
[17]Ye R.W,Averill B.A,Tiedje J.M.Denitrification:Production and consumption of nitricoxide[J].Environ Microbiot,1994,60:281-286
[18]Carr GJ,Page M.D,Fergtlson S.J,et al.The energy-conserving nitric-oxide-reductase system in Paracoccus denitrifican Distrbution,from the nitrite reductase that catalyses synthesis of nitric oxide and evldence from trapping experiments for nitric oxide as a free intermediate during denitrification[J].Eur.J.Biochem,1989,179:686-692
[19]Car G.J,Feruson S.J.The nitric oxide reductase of Paracoccus denitrificans[J].Biochem,1990,269:423-430
[20]Heiss B,Frunzke K,W Zumft.Formation of N=N bond from nitric oxide by a membrane-bound cytochrome bc complex of Nitrate-respiring(denilrifying)Psedomonoas stuteri[J].Bacteriol,1989,171:3288-3297
[21]Heikkila M P,Honisch U,Wunsch P,et al.Role of the tat transport system in nitrous oxide reductase translocation and cytochrome cd1 biosynthesus in Psudomonas stutzeri[J].Journal of Bacteriology,2001,183:1663-1671
[22]殷芳芳,王淑莹,昂雪野等.碳源类型对低温条件下生物反硝化的影响[J].环境科学,2009,30(1):108-113
[23]于爱茸,李尤,俞吉安.一株耐氧反硝化菌的筛选及脱氮特性研究[J].微生物学杂,2005,25(3):77-81
[24]Huang H.K,Tseng SK.Nitrate reduction by a citrobacter diversus under aerobic environment[J].Appl Microbiol Biotechno],2001,55:90-94
[25]殷士学,陈丽敏.土壤中硝化、反硝化微生物的研究进展[J].土壤学报,2000,39:116-125
[26]杨基先,高珊珊,马放等.一株好氧反硝化细菌的分离鉴定及反硝化能力[J].环境科学报,2008,28(7):1301-1307
[27]Katarzyna Bernat,Irena Wojnowska-Baryla.Carbon source in aerobic denitrification[J].Biochemical Engineering Journal,2007,36:l16 -122
[28]曾庆武,梁运祥,葛向阳.反硝化细菌的分离筛选及其反硝化特性的初步研究[J].华中农业大学学报,2008,27(5):617-620
[29]Charles GIass,Joann silver.Denitrification kinetics of high nitrate concentration water:pH effecton inhibition and nitrite accumulation[J].Wat.Res,1998,32(3):831-839
[30]Liu L.H,Koenig A.Use of limestone for pH control in autotrophic denitrification:batchexperiments[J].Process Biochemistry,2002,37:885-893
[31]谢珊,李小明,曾光明等.同步硝化反硝化实现途径的探讨[J].环境科学与技术,2004,27(2):107-109.
[32]Alexander M.Introduction to soil microbiology.John Wiley and Sons,NewYork,1997
[33]李树刚,马光庭.生物脱氮菌种筛选及条件选择的研究[J].广西大学学报(自然科学版),2003,28(2):173-176
[34]李卿.好氧反硝化细菌的筛选和生理特性的研究:[硕士学位论文]长春:吉林大学,2006.
[35]阮立权,陈坚.好氧颗粒污泥同步硝化反硝化脱氮过程中N_2O的产生[J].无锡轻工业大学学报,2004,23(4):37-64
[36]Sunjin Hwang,Keisuke Hanaki.Effects of oxygen concentration and moisture content of refuse on nitrification,denitrification and nitrous oxide Production[J].Bioresource Technology,2000,71:159-165
[37]徐亚同.废水生物处理的运行和管理.上海:华东师范大学出版社,1989:181页
[38]Su J J,Liu B Y,Liu C Y.Comparison of aerobic denitrification under high oxygen at mosphereby Thiosphaera pantotropha ATCC35512 and Pseudomonas stutzeri SU2 newly isolated from the actived sludge of a piggery wastewater treatment system[J].Journal of Applied Microbio,2001,90:457-462
[39]Huang H K,Tseng S K.Nitrate reduction by Citrobacter diversus under aerobic environment[J].Appl Microbiol Biotechnol,2001,55:90-94
[40]Patureau D,Zumstein E,Delgenes J P,et al.Aerobic denitrification isolation from diverse natural and managed ecosystems[J].Microbiol Ecol,2000,39:145-152
[41]Lukow T,Diekmann H.Aerobic denitrification by a newly heterotrophic bacterium strain TL1[J].Biotechnol Lett,1997,11(19):1157-1159
[42]Chen F,Xia Q,Ju L K.Aerobic denitrification of Pseudomonas aeruginosa monitored by online NAD(P)H fluorescence[J].Appl and Environ Microbiol,2003,69(11):6715-6722
[43]Kesserli P,Kiss I,Bihari Z,et al.Biological denitrification in a continuous-flow pilot bioreactor containing immobilized Pseudomonas butannovora cells[J].Bioreso Technol,2003,87:75-80
[44]Kim Y J,Yoshizawa M,Takenaka S,et al.Isolation and culture conditions of a Klebsiella pneumoniae strain that can utilize ammonium and nitrate ions simultaneously with controlled iron and molybdate ion concentrations[J].Biosci Biotechnol Biochem,2002,66(5):996-1001
[45]郑平.环境微生物学.浙江:浙江大学出版社,2003
[46]Brierley EDR,Wood M.Heterotrophic nitrification in an acid forest soil:isolation and characterisation of a nitrifying bacterium[J].Soil Biology & Biochemistry,2001,33(10):1403-1409
[47]Schmidt I,Sliekers O,Schmid M,et al.New concepts of microbial treatment processes for the nitrogen removal in wastewater[J].FEMS Microbiology Reviews,2003,27(4):481-492
[48]杨宗政,王鑫,庞金钊等.异养硝化菌的分离及其强化活性污泥脱氮效果[J].中国给水排水, 2006,22(21):67-70
[49]张彤,庞金钊,杨宗政等.异养硝化菌的脱氮研究[J].水科学与工程技术,2006,6:41-42
[50]苟莎,黄钧.异养硝化细菌脱氮特性及研究进展[J].微生物学通报,2009,36(2):255-260
[51]Gupta AB.Thiosphaera pantotropha:a sulphur bacterium capable of simultaneoush eterotrophic nitrification and aerobic denitrification[J].Enzyme and Microbial Technology,1997,21:589-595
[52]何霞,吕剑,何义亮.异养硝化机理的研究进展[J].微生物学报,2006,46(5):844-847
[53]Wehrfritz JM,Carter JP,Spiro S,et al.Hydroxylamine oxidation in heterotrophic nitrate-reducing soil bacteria and purification of a hydroxylamine-cytochrome oxidoreductase from a Pseudomonas species[J].Arch Microbiol,1997,166:421-424
[54]Joo HS,Hirai M,Shoda M.Nitrification and denitrification in high-strength ammonium by Alcaligenes faecalis[J].Biotechnology Letters,2005,27(11):773-778
[55]Hu TL,Kung KT.Study of heterotrophic nitrifying bacteria from wastewater treatment systems treating acrylonitrile,butadiene and styrene resin wastewater[J].Water Science and Technology 2000,42(3-4):315-321
[56]陈赵芳,尹立红,浦跃朴等.一株异养硝化菌的筛选及其脱氮条件[J].东南大学学报(自然科学版),2007,37(3):486-490
[57]Su JJ,Yeh KS,Tsheng PW.A strain of Pseudomonas sp.isolated from piggery wastewater treatment systems with heterotrophic nitrification capability in Taiwan[J].Current Microbiology,2006,53(1):77-81
[58]Kim JK,Park KJ,Cho KS,et al.Aerobic nitrification-denitrification by heterotrophic Bacillus strains[J].Bioresource Technology,2005,96(17):1897-1906
[59]何霞,赵彬,吕剑等.异养硝化细菌Bacillus sp.LY脱氮性能研究[J].环境科学,2007,28(6):1404-1408
[60]Blagodatsky SA,Kesik M,Papen H,et al.Production of NOand N_2O by the heterotrophic nitrifier Alcaligenes faecalis subsp,parafaecalis under varying conditions of oxygen saturation[J].Geomicrobiology Journal,2006,23(3-4):165-176
[61]曹喜涛,常志州,黄红英等.1株高温异养硝化细菌的分离鉴定和特性研究[J].安徽农业科学2006,34(19):4833-4834
[62]林燕,何义亮,李春杰等.MBR同步硝化反硝化及异养硝化试验研究[J].环境科学与技术,2006,29(1):7-9
[63]Anshuman A,Khardenavis,Atya Kapley,et al.Simultaneous nitrification and denitrification by diverse Diaphorobacter sp[J].Appl Microbiol Biotechnol,2007,77:403-409
[64]林燕,孔海南,何义亮等.异养硝化细菌的分离及其硝化特性实验研究[J].环境科学,27(2):324-328
[65]Hung-Soo Joo,Mitsuyo Hirai,Makoto Shoda.Characteristics of Ammonium Removal by Heterotrophic Nitrification-Aerobic Denitrification by Alcaligenes faecalis No.4[J].Journal of Bioscience and Bioengineering.2005,100(2):184-191
[66]刘正.高浓度含盐废水生物处理技术[J].化工环保,2004,24(增刊):209-211
[67]东秀珠,蔡妙英等.常见细菌系统鉴定手册.北京:科学出版社,2001:62-65
[68]R.E布坎南,N.E吉本斯等编.伯杰细菌鉴定手册(第八版).科学出版社,1984
[69]王弘宇,马放,苏俊峰等.好氧反硝化菌株的鉴定及其反硝化特性研究[J].环境科学.2007,28(7):1548-1552
[70]孔庆鑫.一株新型的脱氮微生物的分离鉴定及其脱氮机制.城市环境与城市生态[J].2005,13(2):36-39
[71]崔家荣.水中氨氮纳氏试剂分光光度法测定[J].现代农业科技,2008,8:208-209
[72]国家环境保护总局和《水和废水监测分析方法》编委会编.水和废水监测分析方法.北京:中国环境科学出版社,271-274
[73]孙仕萍,邢大荣,张岚等.水中硝酸盐氮的麝香草酚分光光度测定法[J].环境与健康杂志,2007,24(4):256-257
[74]Chun-ChinWang,Chi-Mei Lee.Isolation of the caprolactam denitrifying bacteria from a Wastewater treatment system manufactured with acrylonitrile-butadiene-styrene resin[J].Journal of Hazardous Materials 2007,145:136-141
[75]苏俊峰,王继华,马放等.好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究[J].环境科学2007,28(10):2332-2335
[76]HAMID-REZA KARIMINIAAE-HAMEDAANI,KOHZO KANDA,FUMIO KATO.Denitrification Activity of the Bacterium Pseudomonas sp.ASM-2-3 Isolated from the Ariake Sea Tideland[J].Journal of Bioscience and Bioengineering,2004,97(1),39-44
[77]项慕飞.好氧反硝化菌的分离筛选和鉴别研究:[硕士学位论文]北京:北京工商大学,2007
[78]刘水平,罗志勇.琼脂糖凝胶电泳实验技巧[J].实用预防医学,2006,13(4):1068-1069
[79]Kesser U P,Kiss I,Bihari Z,et al.Biological denitrificationin a continuous 2 flow pilot bioreactor containing immobilized Pseudomonas butnnovora cells[J].Bioresource Technology,2003,87:75-80
[80]Takaya N,Maria A ntoninaB C S,Yasushi S,et al.Aerobic denitrification bacteria that produce low levels of nitrous oxide[J].App and Envir Microbio,2003,69(6):3152-3157.
[82]燕永亮,杨剑,陈立宏等.斯氏假单胞菌(Pseudomonas stutzeri)A1501反硝化相关基因结构 及功能分析[J].中国科学C辑,2005,35(3):246-253
[83]周丽华,陈士超,邓志瑞等.太湖沉积物中的可培养细菌:Ⅰ.细菌多样性初步分析[J].湖泊科学,2009,21(1):27-35
[84]Irene vonder Weid,Joana M.Marques,Claudia D.Cunha,et al.Identification and biodegradation potential of a novel strain of Dietzia cinnamea isolated from a petroleum-contaminated tropical soil[J].Systematic and Applied Microbiology,2007,30:331-339
[85]刘真,邵宗泽.南海深海沉积物烷烃降解菌的富集分离与多样性初步分析[J].微生物学报,2007,47(5):869-873
[86]A.A.Shapovalova,T.V.Khijniak,T.P.Tourov,et al.Heterotrophic denitrification at extremely high salt and pH by haloalkaliphilic Gammaproteobacteria from hypersaline soda lakes[J].Extremophiles,2008,12:619-625
[87]Sachiko Yoshie,Hiroshi Makino,Hidenobu Hirosawa et al.Molecular analysis of halophilic bacterial community for high-rate denitrification of saline industrial wastewater[J].Appl Microbiol Biotechnol,2006,72:182-189
[88]宁卓,张波.嗜盐菌的研究进展及应用[J].苏盐科技,2007,3(1):31-32
[89]王振雄,徐毅,周培瑾.嗜盐碱古生菌新种的系统分类学研究[J].微生物学报,2000,40(2):115-120
[90]马放,周丹丹,王宏宇等.一株好氧反硝化细菌生理生态特性的研究[J].哈尔滨工业大学学报,2006,38(4):575-577
[91]P.Elefsiniotis,D.Li.The effect of temperature and carbon source on denitrification using volatile fatty acids[J].Biochemical Engineering Journal,2006,28:148-155
[92]杨莎莎,宋英豪,赵宗升等.pH值和碳氮比对亚硝酸型反硝化影响的研究[J].环境工程学报,2007,12(1):15-19
[93]闫博.高效反硝化菌的分离及其在水体中脱氮规律的研究:[硕士学位论文]天津:天津大学,2002
[94]赵庆.破源浓度对反硝化聚磷效果的影响研究:[硕士学位论文]西安:西安建筑科技大学,2003
[95]Hung-Soo Joo,Mitsuyo Hirai,Makoto Shoda.Piggery wastewater treatment using Alcaligenes faecalis strain No.4 with heterotrophic nitrification and aerobic denitrification[J].Water Research,2006,(40):3029-3036
[96]Reto Manser,Willi Gujer,Hansruedi Siegrist.Oecay processes of nitrifying bacteria in biological wastewater treatment systems.Water Research,2006,(40):2416-2426
[97]苏俊峰.异养型同步硝化反硝化脱氮技术及微生物特性研究:[工学博士学位论文].哈尔滨: 哈尔滨工业大学,2007
[98]Soren Rysgaard,Peter Thastu M,Tage Dalsgaard.Effects of Salinityon NH_4~+-adsorption capacity,Nitrification,and Denitrification in Danish Estuarine Sediments.Estuaries,1999,22(3):21-30
[99]柯国华,汪苹,杨志.好氧反硝化脱氮气态中间产物的研究分析[J].环境科学与技术,2006,29(7):45-54
[100]汪苹,王斌,刘军.同步硝化反硝化处理氨氮废水过程中气态脱氮产物的研究[J].环境科学研究,2005,18(5):67-71