【作者】 王萍；

【导师】 王呈玉；

【作者基本信息】 吉林农业大学 ， 环境工程（专业学位）， 2013， 硕士

【摘要】 石油促进了世界的经济社会发展,促进了人类文明的进步,成为了人类赖以生存的最主要能源之一,其在油田钻采及运送流程中,由于各种客观及主观因素,导致了石油烃类物质的溢出和排放,严重污染土壤资源及水资源,甚至污染大气环境,由于石油污染物质难清理,难降解等问题,给人类和生态系统带来巨大的危害,严重威胁到人类和其他生物的生存和发展,所以石油污染问题已经备受世界各地学者的关注。本文菌株是从持久受原油污染的土壤中筛选得到四株原油降解菌,编号为C-1、C-2、C-3和C-4,采用紫外分光光度法测定菌株的原油降解率,测定OD(600nm)的方式体现菌株在不同条件下的生长状况,测定MATH值和排油圈直径等了解菌株的性能,主要结论如下：1.单一变量法：通过测定不同条件下的初始原油浓度,氯化钠浓度,不同碳源,营养物质等情况下微生物的降解率,得知以上四种菌株的最适生长条件分别为：C-1菌株的最适生长条件为C(初始原油浓度)=1000mg/l,pH(初始值)=7,盐浓度10%,温度为25℃,C-2菌株的最适生长条件为C(初始原油浓度)=800mg/l,pH(初始值)=7,盐浓度10%,温度为25℃,C-3菌株的最适生长条件为C(初始原油浓度)=600mg/l,pH (初始值)=7,盐浓度12%,温度为30℃,C-4菌株的最适生长条件为C(初始原油浓度)=800mg/l,pH(初始值)=7,盐浓度12%,温度为25℃,且在以上最优的条件下微生物降解能力大小顺序为：C-4>C-2>C-1>C-3。2.微生物对碳源利用能力有所不同,通过测定菌株生长状况,得知其利用碳源能力的难易排序为：链烷烃<多环烷烃<芳香烃类,且其中链烷烃是最容易被微生物降解利用的,其中C-1,C-2菌株对于降解苯酚,硝基苯及甲苯等苯酚类碳源的能力明显强于C-3和C-4,而C-3,C-4菌株降解链烷烃的能力明显强于C-1,C-2菌株。3.混合菌的降解率明显高于单一菌的降解率,两种菌相互混合后,随着处理时间的延长,原油的降解率都有所升高,在前期混合菌的优势并不明显,后期,混合菌的降解优势日益显现,结果说明三种菌株相互间既没有较强的共生关系,同样也不存在拮抗作用,通过降解率的测定,可以看出三种高效菌混合比任何独立菌柱和两种混合菌的降解效果好。4.比较牛肉膏蛋白胨培养液和柴油无机盐培养液中菌株的生长情况,说明菌株对亲水性碳源的利用能力较强,及通过排油圈直径的测定结果显示,四种菌株均能产生表面活性物质,C-3菌的油圈直径最大,说明其产生的表面活性物质最多,采用MATH法对菌体表面亲油性进行分析得到,其亲油的能力为:C-4>C-2>C-1>C-3。更多还原

【Abstract】 Petroleum Petroleum has become one of the most important energy sources for human survival and development for it is promoting the development of the world’s economic social and human civilization. Some objective and subjective factors leads to petroleum hydrocarbons pilling over and emission,which pollutes soil resources and water resources seriously, even the air. Hence petroleum pollution brings great harm to humans and environment, and it has threatened the survival and development of humans and other organisms for its hard degradation. The problem of petroleum pollution has attracted the attention of scientists around the world.In this study, C-1、C-2、C-3和C-4Four Petroleum degrading bacteria Strains are screened from the polluted soil by crude oil.We determined the rate of oil degradation using ultraviolet spectrophotometer, observed the Growth of the strains in different conditions by determining their OD(600nm). Understanded the strain properties by determining their MATH value and exclusive circle diameters The main conclusions are as follows:Through the determination of the initial concentration of crude oil under different conditions, different concentrations of sodium chloride, carbon source, nutrient condition microbial degradation rate,1. Change the single factor such as the concentration of crude oil and different concentrations of sodium chloride, carbon source, nutrient condition and so on to measure microbial degradation rate,to choose the best conditions, the result is:the concentration of crude oil is1000mg/l, the concentration of salt is10%,the temperature is25℃and pH=7, these are the best conditions for strain C-1., The concentration of crude oil is800mg/l, the concentration of salt is10%,the temperature is25℃and pH=7, these are the best conditions for strain C-2, The concentration of crude oil is600mg/l, the concentration of salt is12%,the temperature is30℃and pH=7, tthese are the best conditions for strain C-3, The concentration of crude oil is800mg/l, the concentration of salt is12%,the temperature is25℃and pH=7, these are the best conditions for strain C-4, In those conditions, the order of Microbial degradation ability is C-4>C-2>C-1>C-3.2.Through the determination of strain growth conditions for Microbial carbon utilization ability was different, we knew the order of microorganism’s ability of using carbon was: aliphatic hydrocarbon>aromatic hydrocarbon> PAHs. Strains C-l and C-2was stronger than strains C-3and C-4to degradate benzene compounds,.and strains C-3and C-4was stronger than strains C-1and C-2to degradate alkane.3. Degradation rate of mixed bacteria was significantly higher than that of single strain, and with the time going the rate of degradation were rise which was not odvious five days ago, but ten days later. The results showed that there was not strong symbiotic relationship but also no no antagonism among three strains. By measuring the degradation rate,it can be seen that the mixing of three kinds of high efficient bacteria degradation performance than any independent bacteria column and two kinds of mixed bacteria make the degradation better than any independent bacteria and two kinds of mixed bacteria4. Made the strains grow in beef extract peptone culture solution and diesel inorganic salt medium and compared theirs growth rate, we knew Strains utilize the hydrophilic carbon source effectively. The measurement of the oil drain coil diameter indicated that all four strains could produce surfactant, oil ring diameters of C-3bacteria was bigger which indicated that C-3bacteria produced the most surface active substance.The surface hydrophobicity of bactera were analysed by the MATH method, its lipophilic capacity was:C-4>C-2>C-1>C-3更多还原