【作者】 孙伟；

【导师】 马放； 杨基先；

【作者基本信息】 哈尔滨工业大学 ， 微生物学， 2008， 硕士

【摘要】 目前,我国多数油田已进入高含水期,这将对原油的集输管线、回注管线及污水处理系统带来严重的腐蚀危害。在油田注水系统中,硫酸盐还原菌(SRB)是微生物腐蚀(MIC)的主要因素之一。微生物腐蚀广泛存在于土壤?海水供水系统及油田环境,而且已经造成了程度不同的腐蚀破坏?据统计,许多严重的腐蚀破坏,70%～80%直接由细菌引起或者与细菌有关,特别是硫酸盐还原菌。本课题改良了传统的厌氧培养方法;改良了硫酸盐还原菌、反硝化细菌的选择性培养基;开发了同步反硝化硫酸盐还原菌的选择性培养基;运用原子力显微镜对菌株进行形态观察,运用16SrDNA和脂肪酸鉴定方法对菌株进行系统发育分析,采用离子色谱方法记录培养液中的NO3-、NO2-和SO42-浓度,对菌株进行功能验证;对几株分离的硫酸盐还原菌进行了聚丙烯酰胺降解特性的测定。分离得到三株硫酸盐还原菌H1、I7和I8。经生理生化鉴定得出,H1为短杆芽孢杆菌,严格厌氧;经16SrDNA和脂肪酸鉴定得出,该菌株为梭杆菌,暂命名为Clostridium Clostridium bifermentans H1;经测定,其对聚丙烯酰胺的去除率为30.8%。对I7菌株的鉴定方法基本同上,所得出的结论为该菌为短杆无芽孢菌,严格厌氧,初步鉴定为Enterobacter中的新种,暂命名为Enterobacter cloacae I7,属于一般丙烯酰胺低聚体的衍生物。I8菌株也为短杆菌,兼性厌氧,初步鉴定为肠杆菌中的一个新种,暂命名为Enterobacter HPAMDegraded Bacteria I8。分离得到三株反硝化细菌F1、F18和F22-1,均为革兰氏阴性菌,在严格厌氧环境下生长速度较硫酸盐还原菌稍慢。经16SrDNA和脂肪酸鉴定得出,三株菌分别为Clostridium butyricum F1,对NO3-去除率为39.17%;Clostridium sp. F18,对NO3-去除率为58.56%;Thauera sp. F22-1,对NO3-去除率为44.56%。分离得到三株同步反硝化硫酸盐还原菌SN10,SN9,SN22-2,初步鉴定三株菌分别为Paenibacillus sp. SN10,对NO3-的去除率为98.52%,对SO 42-的去除率为96.49%;Paenibacillus lautus SN9,对NO3-的去除率为97.76%,对SO 42-的去除率为96.22%;Bacillus coagulans SN22-2,对NO3-的去除率为97.98%,对SO 42-的去除率为96.48%。更多还原

【Abstract】 At present, the majority of our oilfield has entered a high moisture content period, which would cause the oil pipeline, the inject pipeline and sewage treatment systems have serious corrosion hazards. In oilfield water injection system, sulfate-reducing bacteria (SRB) is one of the main factors of microbiologically influenced corrosion (MIC). MIC widespread in soil and water supply systems and oilfield environment, and has caused varying degrees of corrosion damage. According to statistics, 70% to 80% of a number of serious corrosion damages were caused directly by bacteria or have relevance with bacteria, in particular to sulfate-reducing bacteria (SRB).The study improved the traditional anaerobic training methods, improved the selective medium of the sulfate-reducing bacteria and the denitrifying bacteria, developed the selective medium of the denitrification and sulfate-reducing bacteria simultaneously, used atomic force microscope for the observation to the strains, used the 16SrDNA and the fatty acid identification of the strain to take a systematic analysis, used the ion chromatography to measure the concentration of NO3-, NO2-and SO42-, took the strain for functional verification, and measured the polyacrylamide degradation characteristics.Three sulfate-reducing bacteria H1, I7 and I8 were isolated. It can be known that, after the physiological and biochemical identification, H1 is a short-Bacillus, strictly anaerobic. After the identification of 16SrDNA and fatty acids, the strains of Fusobacterium tentatively were named by Clostridium Clostridium bifermentans H1. After the determination, the removal rate of polyacrylamide is 30.8%. The same identification method was taken with I7 strain. The bacteria is short Bacillus without spore, strictly anaerobic, identified as Enterobacter of new species, named for Enterobacter cloacae I7, a general acrylamide oligomer derivatives. I8 is short Bacillus strains, facultative anaerobic, identified as Enterobacteriaceae of a new species, named Enterobacter HPAMDegraded Bacteria I8 with the removal rate 63.17% .Three denitrifying bacteria F1, F18 and F22-1 were isolated. With the same identifying method above, the strains are all gram-negative bacteria, strictly anaerobic, the growth rate is a little slower than the rate of sulfate-reducing bacteria. After 16 SrDNA and fatty acid identification, three bacteria were gained: Clostridium butyricum F1, whose nitrate removal rate reached 39.17%. Clostridium sp. F18, nitrate removal rate of 58.56%. Thauera sp. F22-1, nitrate removal rate reached 44.56%.Three denitrifying and sulfate-reducing bacteria simultaneously SN10, SN9 and SN22-2 were isolated. After the preliminary identification, the strains are Paenibacillus sp. SN10, the NO3- removal rate reached 98.52 percent, SO42- removal rate was 96.49%. Paenibacillus lautus SN9, the NO3- removal rate reached 97.76 percent, SO42- removal rate was 96.22%. Bacillus coagulans SN22-2, the NO3- removal rate reached 97.98%, SO42- removal rate was 96.48%.更多还原