Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin 300071, PR China; College of Life Sciences, Nankai University, Tianjin 300071, PR China.
Research Institute of Experiment and Detection, Xinjiang Oilfield Company, Karamay 834000, PR China.
J Biosci Bioeng. 2014 Feb;117(2):215-221. doi: 10.1016/j.jbiosc.2013.07.009. Epub 2013 Aug 19.
Compared to medium-high temperature petroleum reservoirs (30°C-73°C), little is known about microbial regulation by nutrients in low-temperature reservoirs. In this study, we report the performance (oil emulsification and biogas production) and community structure of indigenous microorganisms from a low-temperature (22.6°C) petroleum reservoir during nutrient stimulation. Culture-dependent approaches indicated that the number of hydrocarbon-oxidizing bacteria (HOB), nitrate-reducing bacteria (NRB) and methane-producing bacteria (MPB) increased by between 10- and 1000-fold, while sulfate-reducing bacteria (SRB) were observed at low levels during stimulation. Phylogenetic analysis of the 16S rRNA gene indicated that Pseudomonas, Ochrobactrum, Acinetobacter, Halomonas and Marinobacter, which have the capability to produce surfactants, were selectively enriched. Methanoculleus, Methanosaeta, Methanocorpusculum and Methanocalculus showed the largest increase in relative abundance among archaea. Micro-emulsion formed with an average oil droplet diameter of 14.3 μm (ranging between 4.1 μm and 84.2 μm) during stimulation. Gas chromatographic analysis of gas production (186 mL gas/200 mL medium) showed the levels of CO2 and CH4 increased 8.97% and 6.21%, respectively. Similar to medium-high temperature reservoirs, HOB, NRB, SRB and MPB were ubiquitous in the low-temperature reservoir, and oil emulsification and gas production were the main phenomena observed during stimulation. Oil emulsification required a longer duration of time to occur in the low-temperature reservoir.
与中高温油藏（30°C-73°C）相比，低温油藏中微生物受营养物质调控的情况知之甚少。在本研究中，我们报告了来自低温（22.6°C）油藏中土著微生物在营养刺激下的性能（油乳化和沼气生成）和群落结构。基于培养的方法表明，烃氧化菌（HOB）、硝酸盐还原菌（NRB）和产甲烷菌（MPB）的数量增加了 10-1000 倍，而硫酸盐还原菌（SRB）在刺激过程中数量较低。16S rRNA 基因的系统发育分析表明，具有产生表面活性剂能力的假单胞菌、氧化亚铁硫杆菌、不动杆菌、盐单胞菌和海洋杆菌被选择性富集。古菌中甲烷微球菌、产甲烷丝菌、产甲烷球菌和产甲烷八叠球菌的相对丰度增加最大。刺激过程中形成的微乳液平均油滴直径为 14.3μm（范围为 4.1μm 至 84.2μm）。气体产生的气相色谱分析（200mL 培养基中产生 186mL 气体）表明 CO2 和 CH4 的水平分别增加了 8.97%和 6.21%。与中高温油藏类似，HOB、NRB、SRB 和 MPB 在低温油藏中普遍存在，油乳化和气体产生是刺激过程中观察到的主要现象。油乳化在低温油藏中需要更长的时间才能发生。
