Nitrate and oxygen significantly changed the abundance rather than structure of sulphate-reducing and sulphur-oxidising bacteria in water retrieved from petroleum reservoirs.

Sulphate-reducing bacteria (SRB) are the main culprits of microbiologically influenced corrosion in water-flooding petroleum reservoirs, but some sulphur-oxidising bacteria (SOB) are stimulated when nitrate and oxygen are injected, which control the growth of SRB. This study aimed to determine the distributions of SRB and SOB communities in injection-production systems and to analyse the responses of these bacteria to different treatments involving nitrate and oxygen. Desulfovibrio, Desulfobacca, Desulfobulbus, Sulfuricurvum and Dechloromonas were commonly detected via 16S rRNA gene sequencing. Still, no significant differences were observed for either the SRB or SOB communities between injection and production wells. Three groups of water samples collected from different sampling sites were incubated. Statistical analysis of functional gene (dsrB and soxB) clone libraries and quantitative polymerase chain reaction showed that the SOB community structures were more strongly affected by the nitrate and oxygen levels than SRB clustered according to the sampling site; moreover, both the SRB and SOB community abundances significantly changed. Additionally, the highest SRB inhibitory effect and the lowest dsrB/soxB ratio were obtained under high concentrations of nitrate and oxygen in the three groups, suggesting that the synergistic effect of nitrate and oxygen level was strong on the inhibition of SRB by potential SOB.

Bacterial and Archaeal Community Distribution in Oilfield Water Re-injection Facilities and the Influences from Microorganisms in Injected Water.

Water flooding is widely employed for oil production worldwide. However, there has never been a systematic investigation of the microbial communities occurring in oilfield water re-injection facilities. Here, we investigated the distribution of bacterial and archaeal communities in water re-injection facilities of an oilfield, and illustrated the combined influences of environmental variation and the microorganisms in injected water on the microbial communities. Bacterial communities from the surface injection facilities were dominated by aerobic or facultative anaerobic Betaproteobacteria, Alphaproteobacteria, and Flavobacteria, whereas Clostridia, Deltaproteobacteria, Anaerolineae, and Synergistia predominated in downhole of the injection wells, and Gammaproteobacteria, Betaproteobacteria, and Epsilonproteobacteria predominated in the production wells. Methanosaeta, Methanobacterium, and Methanolinea were dominant archaea in the injection facilities, while Methanosaeta, Methanomethylovorans, and Methanoculleus predominated in the production wells. This study also demonstrated that the microorganisms in injected water could be easily transferred from injection station to wellheads and downhole of injection wells, and environmental variation and diffusion-limited microbial transfer resulted from formation filtration were the main factors determining microbial community assembly in oil-bearing strata. The results provide novel information on the bacterial and archaeal communities and the underlying mechanisms occurring in oilfield water re-injection facilities, and benefit the development of effective microbiologically enhanced oil recovery and microbiologically prevented reservoir souring programs.

Low-Abundance Dietzia Inhabiting a Water-Flooding Oil Reservoir and the Application Potential for Oil Recovery.

With the development of molecular ecology, increasing low-abundance microbial populations were detected in oil reservoirs. However, our knowledge about the oil recovery potential of these populations is lacking. In this study, the oil recovery potential of low-abundance Dietzia that accounts for less than 0.5% in microbial communities of a water-flooding oil reservoir was investigated. On the one hand, Dietzia sp. strain ZQ-4 was isolated from the water-flooding reservoir, and the oil recovery potential was evaluated from the perspective of metabolisms and oil-displacing test. On the other hand, the strain has alkane hydroxylase genes alkB and P450 CYP153 and can degrade hydrocarbons and produce surfactants. The core-flooding test indicated that displacing fluid with 2% ZQ-4 fermentation broth increased 18.82% oil displacement efficiency, and in situ fermentation of ZQ-4 increased 1.97% oil displacement efficiency. Furthermore, the responses of Dietzia in the reservoir accompanied by the nutrient stimulation process was investigated and showed that Dietzia in some oil production wells significantly increased in the initial phase of nutrient injection and sharply decreased along with the continuous nutrient injection. Overall, this study indicates that Dietzia sp. strain has application potential for enhancing oil recovery through an ex situ way, yet the ability of oil recovery in situ based on nutrient injection is limited.

Spatial isolation and environmental factors drive distinct bacterial and archaeal communities in different types of petroleum reservoirs in China.

To investigate the spatial distribution of microbial communities and their drivers in petroleum reservoir environments, we performed pyrosequencing of microbial partial 16S rRNA, derived from 20 geographically separated water-flooding reservoirs, and two reservoirs that had not been flooded, in China. The results indicated that distinct underground microbial communities inhabited the different reservoirs. Compared with the bacteria, archaeal alpha-diversity was not strongly correlated with the environmental variables. The variation of the bacterial and archaeal community compositions was affected synthetically, by the mining patterns, spatial isolation, reservoir temperature, salinity and pH of the formation brine. The environmental factors explained 64.22% and 78.26% of the total variance for the bacterial and archaeal communities, respectively. Despite the diverse community compositions, shared populations (48 bacterial and 18 archaeal genera) were found and were dominant in most of the oilfields. Potential indigenous microorganisms, including Carboxydibrachium, Thermosinus, and Neptunomonas, were only detected in a reservoir that had not been flooded with water. This study indicates that: 1) the environmental variation drives distinct microbial communities in different reservoirs; 2) compared with the archaea, the bacterial communities were highly heterogeneous within and among the reservoirs; and 3) despite the community variation, some microorganisms are dominant in multiple petroleum reservoirs.

Microbial abundance and community composition influence production performance in a low-temperature petroleum reservoir.

Enhanced oil recovery using indigenous microorganisms has been successfully applied in the petroleum industry, but the role of microorganisms remains poorly understood. Here, we investigated the relationship between microbial population dynamics and oil production performance during a water flooding process coupled with nutrient injection in a low-temperature petroleum reservoir. Samples were collected monthly over a two-year period. The microbial composition of samples was determined using 16S rRNA gene pyrosequencing and real-time quantitative polymerase chain reaction analyses. Our results indicated that the microbial community structure in each production well microhabitat was dramatically altered during flooding with eutrophic water. As well as an increase in the density of microorganisms, biosurfactant producers, such as Pseudomonas, Alcaligenes, Rhodococcus, and Rhizobium, were detected in abundance. Furthermore, the density of these microorganisms was closely related to the incremental oil production. Oil emulsification and changes in the fluid-production profile were also observed. In addition, we found that microbial community structure was strongly correlated with environmental factors, such as water content and total nitrogen. These results suggest that injected nutrients increase the abundance of microorganisms, particularly biosurfactant producers. These bacteria and their metabolic products subsequently emulsify oil and alter fluid-production profiles to enhance oil recovery.

Dynamic processes of indigenous microorganisms from a low-temperature petroleum reservoir during nutrient stimulation.

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.

Nutrients and oxygen alter reservoir biochemical characters and enhance oil recovery during biostimulation.

Biostimulation of petroleum reservoir to improve oil recovery has been conducted in a large number of oilfields. However, the roles and linkages of organic nutrients, inorganic salts and oxygen content during biostimulation have not been effectively elucidated. Therefore, we investigated the relationships between carbon source, nitrogen source, phosphorus source, oxygen content, and microbial stimulation, oil emulsification, and oil degradation. The organic nutrients (molasses) accelerated microbial growth, and promoted oil emulsification under aerobic conditions. The added molasses also promoted metabolites production (CO2, CH4 and acetic acid) and microbial anaerobic hydrocarbon degradation under anaerobic conditions. (NH4)2HPO4 improved gases production by neutralizing the acidic production and molasses. NaNO3 could also improve gases production by inhibiting sulfate-reducing bacteria to adjust pH value. Oxygen supply was necessary for oil emulsification, but bountiful supply of oxygen aggravated oil degradation, leading the entire ranges of alkanes and some aromatic hydrocarbons were degraded. Core-flooding experiments showed an oil displacement efficiency of 13.81 % in test with air package injected, 8.56 % without air package injection, and 4.77 % in control test with air package injection and 3.61 % without air package injection. The results suggest that the combined effect of organic nutrients, inorganic salts and oxygen content determines microbial growth, while production of metabolites, oil emulsification and biodegradation alter the reservoir biochemical characters and influence oil recovery during stimulation.

[Research on population structure and distribution characteristic of indigenous microorganism in post-polymer-flooding oil reservoir].

Denaturing gradient gel electrophoresis (DGGE) method and principal component analysis (PCA) method were used to analyze the structures of microorganism population in injection wells and production wells of a post-polymer-flooding oil reservoir in Daqing oil field. The results showed that the dominant species in injection wellhead were aerobic bacteria Pseudomonas and Acinenobacter. Facultative anaerobic bacteria Enterbacter was the dominant bacteria in near area of injection wells. Bacteria detected in production wells included Thauera, Clostridia, Pseudomonas, Petrobacter and some uncultured bacteria. Methanosaeta turned out to be the only archaea detected in injection wells, which was an aceticlastic methane-producing archaeon. Archaea detected in production wells consisted of Methanomicrobium, Methanospirillum and Methanobacterium. In general, aerobic bacteria, facultative anaerobe, and strictly anaerobic bacteria distributed successively from injection wells to production wells in this block. The dominant populations of archaea were different between injection wells and production wells, while were influenced by different environments and microbial metabolism products.

[Microbial metabolic characteristics and ecological controlling in petroleum reservoir--a review].

Petroleum reservoir has a variety of microbes with diverse metabolic characteristics and great diversity. These microbes play an important role in geochemical cycle. Research on their metabolism and ecological relationships enables the understanding of Microbial Enhanced Oil Recovery (MEOR). Based on the present researches, we review the microbial metabolic characteristics and ecological relationships in oil reservoir.