Petroleum Microbiology Research Group, Department of Biological Sciences, University of Calgary, 2500 University Dr. NW, Calgary, Alberta T2N 1N4, Canada.
Appl Environ Microbiol. 2010 Aug;76(15):4977-87. doi: 10.1128/AEM.00596-10. Epub 2010 Jun 18.
Community analysis of a mesothermic oil field, subjected to continuous field-wide injection of nitrate to remove sulfide, with denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes indicated the presence of heterotrophic and sulfide-oxidizing, nitrate-reducing bacteria (hNRB and soNRB). These reduce nitrate by dissimilatory nitrate reduction to ammonium (e.g., Sulfurospirillum and Denitrovibrio) or by denitrification (e.g., Sulfurimonas, Arcobacter, and Thauera). Monitoring of ammonium concentrations in producing wells (PWs) indicated that denitrification was the main pathway for nitrate reduction in the field: breakthrough of nitrate and nitrite in two PWs was not associated with an increase in the ammonium concentration, and no increase in the ammonium concentration was seen in any of 11 producing wells during periods of increased nitrate injection. Instead, ammonium concentrations in produced waters decreased on average from 0.3 to 0.2 mM during 2 years of nitrate injection. Physiological studies with produced water-derived hNRB microcosms indicated increased biomass formation associated with denitrification as a possible cause for decreasing ammonium concentrations. Use of anammox-specific primers and cloning of the resulting PCR product gave clones affiliated with the known anammox genera "Candidatus Brocadia" and "Candidatus Kuenenia," indicating that the anammox reaction may also contribute to declining ammonium concentrations. Overall, the results indicate the following: (i) that nitrate injected into an oil field to oxidize sulfide is primarily reduced by denitrifying bacteria, of which many genera have been identified by DGGE, and (ii) that perhaps counterintuitively, nitrate injection leads to decreasing ammonium concentrations in produced waters.
对一个中温油田进行群落分析,该油田连续进行全场范围内的硝酸盐注入以去除硫化物,用聚合酶链反应(PCR)扩增 16S rRNA 基因的变性梯度凝胶电泳(DGGE)表明存在异养和硫氧化、硝酸盐还原菌(hNRB 和 soNRB)。这些细菌通过异化硝酸盐还原为铵(例如 Sulfurospirillum 和 Denitrovibrio)或通过反硝化(例如 Sulfurimonas、Arcobacter 和 Thauera)来还原硝酸盐。监测生产井(PW)中的铵浓度表明,反硝化是该油田硝酸盐还原的主要途径:在两口 PW 中,硝酸盐和亚硝酸盐的突破与铵浓度的增加无关,在硝酸盐注入增加期间,11 口生产井中没有一口的铵浓度增加。相反,在硝酸盐注入的两年中,生产水中的铵浓度平均从 0.3 降至 0.2mM。利用生产水衍生的 hNRB 微宇宙进行的生理研究表明,反硝化过程中生物量的增加可能是铵浓度降低的原因。使用专性厌氧氨氧化菌引物并克隆所得 PCR 产物,得到与已知的厌氧氨氧化菌属“Candidatus Brocadia”和“Candidatus Kuenenia”相关的克隆,表明厌氧氨氧化反应也可能导致铵浓度下降。总的来说,结果表明:(i)注入油田以氧化硫化物的硝酸盐主要被反硝化细菌还原,其中许多属已通过 DGGE 鉴定,(ii)反硝化细菌出乎意料地导致生产水中的铵浓度降低。