降解石油污染的微生物分离、培养及其中试生物修复研究
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摘要
本文通过对辽河油田油污土壤中土著菌的培养、筛分和驯化,得到了三种对辽河原油具有良好耐受能力的细菌菌种。通过液相降解辽河原油的试验,确定三种细菌均对辽河原油具有很好的降解性能。本文以多次正交试验的方法优化了三种细菌的发酵培养基配方并考查了摇瓶发酵的操作,优化了三种微生物的培养温度及pH值条件。并根据三种细菌的生长曲线建立了菌体的生长模型。本文将发酵得到的高密度菌液制成菌剂投加到辽河油田油污现场进行的中试试验,经过为期60天的降解过程,中试场地内辽河油田地表油污土壤的含油量大幅下降。并对中试场地土壤中微生物的数量进行分析,分析表明微生物的生长主要受气温的影响,并以数值模拟的方法得到了中试场地自然条件下优势菌种的生长规律。石油降解微生物与油污土壤表面的原油接触是降解过程的必要前提,因此对细菌在土壤中的迁移规律进行研究。本文根据土壤颗粒的结构特性及现代分形方法取得了土壤孔隙结构参数并根据传质理论以简单易行的方法确定了细菌在土壤中扩散系数;根据热力学平衡实验确定了细菌在土壤中的平衡吸附模型。在此基础上提出了微生物在土壤中的迁移模型。通过对迁移影响因素的分析,明确了各参数对迁移所起的重要作用,加深了对细菌迁移规律的认识。
Three oil-resistant strains were identified.They were separated by culturing, domesticating and selecting form the contaminated soil of Liaohe oil field. The separated strains were proven that they have good degradation ability to the crud oil by fluid-phase decomposition tests. In this study, the culture medium as well as the shake flask operation condition, namely the optimum culture temperature and pH, was optimized by Orthogonal Tests. The growth models were also constructed according to the growth curves of the separated strains. A bacterium agent which was made using the three bacteria was added to the oil contaminated soil located in Liaohe oil field. This pilot test indicated that the oil content of the contaminated soil was distinctly reduced about 70% by biodegradation in 60 days. Meanwhile the growing pattern of the predominant bacterium group was obtained based on the amount analysis of the microbo. The contact of the bacteria and the oil-contaminated soil is a necessary precondition of soil remediation, so the transportation law of bacteria was also studied in this paper. The parameter of soil pores was obtained based on the soil granule structure characteristic and fractal theory, then the bacteria diffusion coefficient was obtained by utilizing the membrane cell. The equilibrium absorbtion model of the bacteria in soil was identified by thermodynamics balance experiment, which was the bedrock of the constructed microbo transportation model. By analysing the transportation parameters, contribution of each parameter to the transporttion was dug out, and the understanding transportation law of microbio was deepened.
Three oil-resistant strains were identified.They were separated by culturing, domesticating and selecting form the contaminated soil of Liaohe oil field. The separated strains were proven that they have good degradation ability to the crud oil by fluid-phase decomposition tests. In this study, the culture medium as well as the shake flask operation condition, namely the optimum culture temperature and pH, was optimized by Orthogonal Tests. The growth models were also constructed according to the growth curves of the separated strains. A bacterium agent which was made using the three bacteria was added to the oil contaminated soil located in Liaohe oil field. This pilot test indicated that the oil content of the contaminated soil was distinctly reduced about 70% by biodegradation in 60 days. Meanwhile the growing pattern of the predominant bacterium group was obtained based on the amount analysis of the microbo. The contact of the bacteria and the oil-contaminated soil is a necessary precondition of soil remediation, so the transportation law of bacteria was also studied in this paper. The parameter of soil pores was obtained based on the soil granule structure characteristic and fractal theory, then the bacteria diffusion coefficient was obtained by utilizing the membrane cell. The equilibrium absorbtion model of the bacteria in soil was identified by thermodynamics balance experiment, which was the bedrock of the constructed microbo transportation model. By analysing the transportation parameters, contribution of each parameter to the transporttion was dug out, and the understanding transportation law of microbio was deepened.
引文
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[6]黄国强,姜斌,李鑫钢等. VOCs在土壤孔隙中扩散模型的适用性,天津大学学报, 2004, 37(11):945～948
[7]郑艳梅,李鑫钢,王战强等. AS技术修复MTBE污染地下水的传质研究,农业环境科学学报2005, 24(3):503～50
[8]郑艳梅,黄国强,姜斌等.地下水曝气理论模型研究进展,环境污染与防治, 2006, 28(7):521～525
[9]隋红,李鑫钢,姜斌等.生物通风过程中单井轴对称受污染区域流场的数值模拟,化工学报, 2006, 57(12):2860～2864
[10]杨乐巍,黄国强,李鑫钢.土壤气相抽提(SVE)技术研究进展,环境保护科学, 2006, 32(6):62～65
[11]段云霞,隋红,韩振为等.生物通风修复石油污染土壤的研究进展,环境生态, 2003, 29:25～28
[12]隋红,李鑫钢,黄国强等.土壤有机污染的原位修复技术,环境污染治理技术与设备, 2003, 4(8):41～45
[13]隋红,姜斌,黄国强等.生物通风修复含石油污染物土壤过程,化工学报, 2004, 55(9):1488～1492
[14]黄国强,李鑫钢,徐世民等.土壤气相抽提作用机制探讨和基本数学模型建立,土壤学报, 2004, 41(3):394～400
[15] Leahy J. G. , Microbial degradation of hydrocarbons in the environment, In: Microbial, 1990, 54:305～315
[16]李希明,微生物降解含油废水,重庆环境科学, 1998, 20(1):16～19
[17] Fedorak R. M. , Westlake D. W. , Microbial degradation of aromatics and saturates in Prudhoe Bay crude oil as determined by glass capillary gas chromatography, In: Microbial, 1981, 27:432～443
[18] Killham K. , Amatom, Ladd J. N. , Effect of substrate location in soil and soil pore-water regime on carton turnover. In: Soil Boil Biochem, 1993, 25:57～62
[19] Higgings I. J. et al . Oil Ind. Microb. Ecosyst, Proc Meet, l978, 80～84
[20] Leahy J. G. and Colwell R. R. MicroBial Degradation of Hydrocarbons in Environment .Microbio1.Rev., 1990, 54:305～315.
[21] Bartha R.and Atlas R.M..Transport and Transformations of Petroleum:Biological Processes[R].In:Long-term Environment Effects of Offshore Oil and Gas Development.ed Boesch D F and Rabalais N N.Elsevier Applied Science.New York.1987:287~341.
[22] Texas Research Institute.1984.Laboratory.2001, 10:13～16.
[23] Miller. R. N, Dewney D. C, Carmen V. A. Hinchee R. E, and Leeson A, A summary of bioventing performance at multi—pie air force stites.Broeeedings of the 1993 NWW/A/API scale gasoline spill and venting experiment.Am erican Petroleum In—stitute.Interim Report No.7743~ 5.
[24] Syakti, N. Mazzella, D. Nerini, M. Guiliano, J. C. Bertrand and P. Doumenq. Phospholipid fatty acids of a marine sedimentary microbial community in a laboratory microcosm: Responses to petroleum hydrocarbon contamination Org Geochem , 2006, 37 (11): 1617～1628
[25] M. J. Ayotamuno. Bioremediation of a crude-oil polluted agricultural-soil at Port Harcourt, Nigeria APPL ENERG 2006, 83 (11): 1249～1257.
[26]齐永强,王红旗,郭淼.土壤石油生物降解影响因子正交实验分析.重庆环境科学, 2002, 24(2):29～32
[27]叶淑红,丁鸣,马达,曹方,丁永生,丁德文.微生物修复辽东湾油污染湿地研究.环境科学, 2005, 36(5):143～146
[28] P. Fernández-álvarez, J. Vila, J. M. Garrido-Fernández, M. Grifoll and J. M. Lema. Trials of bioremediation on a beach affected by the heavy oil spill of the Prestige, Hazard Mater, 2006, 137 (3): 1523～1531
[29]李秀艳,魏德洲,何良菊等.氮磷营养盐对微生物降解作用的影响, 2000, 52(4):237～239
[30] Fedorak P. M, Westlake D W, Microbial Degradation of Organic Sulfur Compounds in Prudhoe Bay Crude Oil Canadian. Microbiology, 1983, 29: 291～296
[31] McMillen S. J. , Newland M., In situ bioventing of a diesel fuel spill. In situ and on-site bioremediation, 2002, (4): 297～308
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