Reactive Transport Modeling of Induced Selective Plugging by Leuconostoc Mesenteroides in Carbonate Formations

A process-based mechanistic reactive transport model was developed to understand how in-situ coupled processes and operational factors affect selective plugging of reactive carbonate formations by the fermenting bacteria Leuconostoc mesenteroides that produces a plugging polymer dextran. The growth and transport of L. mesenteroides and the associated (bio) geochemical reactions were simulated explicitly with enzyme activity at the field scale over spatial extents of hundreds of meters. Simulations were performed to explore controls on selective bioplugging of high permeability zones in a representative carbonate reservoir, a process that can be used to improve oil sweep efficiency through lower permeability layers. Simulation results indicate that dextran production and the effectiveness of plugging can be largely affected by sucrose and bacteria injection rates. Selective plugging of high permeability zones can only be achieved when the injection rates are high compared to the rates of dextran production. Otherwise, plugging only occurs at the vicinity of injection wells. Due to the dependence of enzyme activity on pH and the reactive nature of carbonate formations, the chemistry of the injection and the formation water is also important. The injection of sucrose and L. mesenteroides at the optimum pH for dextran production (5.2) leads to the dissolution of calcite and an increase in pH levels. However, the resulting pH does not suppress plugging with dextran. Lactic acid and CO₂ formed during the growth of L. mesenteroides buffers the pH of water to levels between 5.2 and 7.0 for continued dextran production. At neutral and basic pH levels, induced precipitation of calcite does not significantly modify the permeability profile at carbonate concentrations typically found in oilfield formation waters. This is the first work that examines the controlling parameters that affect selective plugging of carbonate formations at the field scale within the context of enhanced oil recovery. The demonstrated approach can be used to identify optimal operational conditions for enhanced oil recovery and other applications where selective plugging can be beneficial.     

Effects of oil reservoir conditions on the production of water‐insoluble Levan by Bacillus licheniformis

Bacillus licheniformis produced a water‐insoluble levan which has potential application as a selective plugging agent in microbial enhanced oil recovery (MEOR). The microorganism grew on sucrose, glucose, and fructose but produced levan only on sucrose. Plugging may thus be selectively controlled in the reservoir by substrate manipulation. B. licheniformis and a crude preparation of its extracellular enzymes were evaluated for their ability to produce levan under reservoir conditions. Oil reservoirs which have a temperature of less than 55°C, a pH between 6 and 9, a pressure less than 500 atm, and a salt concentration of 4% or less are potentially suitable. Examples of such reservoir conditions are found in Lloydminster on the Alberta‐Saskatchewan border, one of the largest Canadian oil reserves.     

Core flooding tests to investigate the effects of IFT reduction and wettability alteration on oil recovery during MEOR process in an Iranian oil reservoir

Microbial enhanced oil recovery (MEOR) refers to the process of using bacterial activities for more oil recovery from oil reservoirs mainly by interfacial tension reduction and wettability alteration mechanisms. Investigating the impact of these two mechanisms on enhanced oil recovery during MEOR process is the main objective of this work. Different analytical methods such as oil spreading and surface activity measurements were utilized to screen the biosurfactant-producing bacteria isolated from the brine of a specific oil reservoir located in the southwest of Iran. The isolates identified by 16S rDNA and biochemical analysis as Enterobacter cloacae (Persian Type Culture Collection (PTCC) 1798) and Enterobacter hormaechei (PTCC 1799) produce 1.53 g/l of biosurfactant. The produced biosurfactant caused substantial surface tension reduction of the growth medium and interfacial tension reduction between oil and brine to 31 and 3.2 mN/m from the original value of 72 and 29 mN/m, respectively. A novel set of core flooding tests, including in situ and ex situ scenarios, was designed to explore the potential of the isolated consortium as an agent for MEOR process. Besides, the individual effects of wettability alteration and IFT reduction on oil recovery efficiency by this process were investigated. The results show that the wettability alteration of the reservoir rock toward neutrally wet condition in the course of the adsorption of bacteria cells and biofilm formation are the dominant mechanisms on the improvement of oil recovery efficiency.     

Response of microbial community structure to microbial plugging in a mesothermic petroleum reservoir in China

Microbial plugging, a microbial enhancement of oil recovery (MEOR) technique, has been applied in a candidate oil reservoir of Daqing Oil Field (China). The goal of this study is to monitor the survival of injected bacteria and reveal the response of microbial communities in field trial of microbial plugging through injection of selected microbial culture broth and nutrients. Culture-dependent enrichment and culture-independent 16S rDNA clone library methods were used. The results show that it was easy to activate targeted biopolymer-producing bacteria in a laboratory environment, and it was difficult for injected exogenous bacteria to survive. In addition, microbial communities in the oil reservoir also changed before and after the field trial. However, microbial communities, activated by fermentative medium for biopolymer-producing bacteria, appeared to show greater differences in the laboratory than in the natural reservoir. It was concluded that microbial populations monitoring was important to MEOR; results of response of microbial communities could provide a guide for the future field trials.     

Swift production of rhamnolipid biosurfactant,biopolymer and synthesis of biosurfactant-wrapped silver nanoparticles and its enhanced oil recovery

Microbial enhanced oil recovery (MEOR) is a kind of enhanced oil recovery (EOR) development, often used as a tertiary stage where oil recovery is no longer possible utilizing primary and secondary conventional techniques. Among a few potential natural operators valuable for MEOR, biosurfactants, biopolymers and biosurfactant based nanoparticles assume key jobs. Biosurfactant which are produced by microorganisms’ act as are surface active agents that can be used as an alternative to chemically synthesized surfactants. Pseudomonas aeruginosa TEN01, a gram-negative bacterium isolated from the petroleum industry is a potential biosurfactant (Rhamnolipid) producer using cassava waste as the substrate. This work focuses on production and characterization of rhamnolipid from P. aeruginosa TEN01 and its use in enhanced oil recovery. The effectiveness of Chitosan that is deacetylated form of chitin which is a biopolymer that provides density and viscosity to the fluids is not known in enhanced oil recovery yet and so it is studied. Moreover, the fabrication of biosurfactant-mediated silver nanocrystals and its application in enhanced oil recovery is also studied. Sand-Pack column was constructed and the mechanism of oil recovery in the column was studied. While incubating the crude oil containing sand packed column with Biosurfactant-biopolymer and brine flooding in the ratio of 1:2, and Biosurfactant incubation - flooding with 3 g/l of biopolymer was found to be 34.28% and 44.5% respectively. The biosurfactant based silver nanoparticles are non-toxic and have better stability when compared to chemically synthesized silver nanoparticles. The oil recovery percentage by chemical based Ag NPs and biosurfactant based Ag NPs are 14.94% and 14.28% respectively.     

Monitoring exogenous and indigenous bacteria by PCR-DGGE technology during the process of microbial enhanced oil recovery

A field experiment was performed to monitor changes in exogenous bacteria and to investigate the diversity of indigenous bacteria during a field trial of microbial enhanced oil recovery (MEOR). Two wells (26-195 and 27-221) were injected with three exogenous strains and then closed to allow for microbial growth and metabolism. After a waiting period, the pumps were restarted and the samples were collected. The bacterial populations of these samples were analyzed by denaturing gradient gel electrophoresis (DGGE) with PCR-amplified 16S rRNA fragments. DGGE profiles indicated that the exogenous strains were retrieved in the production water samples and indigenous strains could also be detected. After the pumps were restarted, average oil yield increased to 1.58 and 4.52 tons per day in wells 26-195 and 27-221, respectively, compared with almost no oil output before the injection of exogenous bacteria. Exogenous bacteria and indigenous bacteria contributed together to the increased oil output. Sequence analysis of the DGGE bands revealed that Proteobacteria were a major component of the predominant bacteria in both wells. Changes in the bacteria population in the reservoirs during MEOR process were monitored by molecular analysis of the 16S rRNA gene sequence. DGGE analysis was a successful approach to investigate the changes in microorganisms used for enhancing oil recovery. The feasibility of MEOR technology in the petroleum industry was also demonstrated.     

Leuconostoc mesenteroides growth kinetics with application to bacterial profile modification

Bacterial profile modification (BPM) is being developed as an oil recovery technique that uses bacteria to selectively plug oil depleted zones within a reservoir to divert displacing fluids (typically water) into oil-rich zones. Leuconostoc mesenteroides, which produces dextran when supplied with sucrose, is a bacterium that is technically feasible for use in profile modification. However, the technique requires controlled bacterial growth to produce selective plugging.A kinetic model for the production of cells and polysaccharides has been developed for L. mesenteroides bacteria. This model, based on data from batch growth experiments, predicts saccharide utilization, cell generation, and dextran production. The underlying mechanism is the extracellular breakdown of sucrose into glucose and fructose and the subsequent production of polysaccharide (dextran). The monosaccharides are then available for growth. Accompanying sucrose consumption is the utilization of yeast extract. The cell requires a complex media that is provided by yeast extract as a source of vitamins and amino acids. Varying the concentration ratio of yeast extract to sucrose in the growth media provides a means of controlling the amount of polymer produced per cell. Consequently, in situ bacteria growth can be controlled by the manipulation of nutrient media composition, thereby providing the ability to create an overall strategy for the use of L. mesenteroides bacteria for profile modification.     

Bacteria in Crude Oil Survived Autoclaving and Stimulated Differentially by Exogenous Bacteria

Autoclaving of crude oil is often used to evaluate the hydrocarbon-degrading abilities of bacteria. This may be potentially useful for bioaugmentation and microbial enhanced oil recovery (MEOR). However, it is not entirely clear if “endogenous” bacteria (e.g., spores) in/on crude oil survive the autoclaving process, or influence subsequent evaluation of the hydrocarbon-degradation abilities of the “exogenous” bacterial strains. To test this, we inoculated autoclaved crude oil medium with six exogenous bacterial strains (three Dietzia strains, two Acinetobacter strains, and one Pseudomonas strain). The survival of the spore-forming Bacillus and Paenibacillus and the non-spore-forming mesophilic Pseudomonas, Dietzia, Alcaligenes, and Microbacterium was detected using a 16S rRNA gene clone library and terminal restriction fragment length polymorphism (T-RFLP) analysis. However, neither bacteria nor bacterial activity was detected in three controls consisting of non-inoculated autoclaved crude oil medium. These results suggest that detection of endogenous bacteria was stimulated by the six inoculated strains. In addition, inoculation with Acinetobacter spp. stimulated detection of Bacillus, while inoculation with Dietzia spp. and Pseudomonas sp. stimulated the detection of more Pseudomonas. In contrast, similar exogenous bacteria stimulated similar endogenous bacteria at the genus level. Based on these results, special emphasis should be applied to evaluate the influence of bacteria capable of surviving autoclaving on the hydrocarbon-degrading abilities of exogenous bacteria, in particular, with regard to bioaugmentation and MEOR. Bioaugmentation and MEOR technologies could then be developed to more accurately direct the growth of specific endogenous bacteria that may then improve the efficiency of treatment or recovery of crude oil.     

厌氧产表面活性剂微生物提高原油采收率的研究进展

微生物强化采油(microbial enhanced oil recovery,MEOR)是近年来在国内外发展迅速的一项提高原油采收率技术。微生物在油藏中高效生产表面活性剂等驱油物质是微生物采油技术成功实施的关键之一。然而,油藏的缺/厌氧环境严重影响好氧表面活性剂产生菌在油藏原位的生存与代谢活性;油藏注空气会增加开采成本,且注入空气的作用时效和范围难以确定。因此,开发厌氧产表面活性剂菌种资源并强化其驱油效率对于提高原油采收率具有重要意义。本文综述了国内外近年来利用厌氧产表面活性剂微生物提高原油采收率的研究进展,简述了微生物厌氧产表面活性剂的相关驱油机理、菌种资源开发现状以及油藏原位驱油应用进展,并对当前的研究提出了一些思考。     

Selection of Bacteria with Favorable Transport Properties Through Porous Rock for the Application of Microbial-Enhanced Oil Recovery

This paper presents a bench-scale study on the transport in highly permeable porous rock of three bacterial species—Bacillus subtilis, Pseudomonas putida, and Clostridium acetobutylicum—potentially applicable in microbial-enhanced oil recovery processes. The transport of cells during the injection of bacterial suspension and nutrient medium was simulated by a deep bed filtration model. Deep bed filtration coefficients and the maximum capacity of cells in porous rock were measured. Low to intermediate (~10⁶/ml) injection concentrations of cellular suspensions are recommended because plugging of inlet surface is less likely to occur. In addition to their resistance to adverse environments, spores of clostridia are strongly recommended for use in microbial-enhanced oil recovery processes since they are easiest among the species tested to push through porous rock. After injection, further transport of bacteria during incubation can occur by growth and mobility through the stagnant nutrient medium which fills the porous rock. We have developed an apparatus to study the migration of bacteria through a Berea sandstone core containing nutrient medium.     

油藏微生物的代谢特征与提高原油采收率技术

油藏是一个高温、高压、少氧、寡营养和封闭的极端环境,油田经过多年注水开发后,在油藏内部形成了相对稳定的微生物群落体系,这些微生物以石油烃分解为起始,形成了一个复杂的食物链,对油藏碳、硫和金属离子的元素地球化学循环起着非常重要的作用。微生物提高原油采收率技术(MEOR)是利用微生物及其代谢产物与油藏和原油发生作用来提高原油采收率的一种新技术,具有成本低、适应性强和环境友好等特点,因此有望成为未来化学驱后油藏和高含水油藏进一步提高采收率的重要手段。对油藏内源微生物及其介导的生化反应,微生物采油原理、发展历程和现场试验进行综述,并提出了未来的发展方向。     

Impact of an indigenous microbial enhanced oil recovery field trial on microbial community structure in a high pour-point oil reservoir

Based on preliminary investigation of microbial populations in a high pour-point oil reservoir, an indigenous microbial enhanced oil recovery (MEOR) field trial was carried out. The purpose of the study is to reveal the impact of the indigenous MEOR process on microbial community structure in the oil reservoir using 16Sr DNA clone library technique. The detailed monitoring results showed significant response of microbial communities during the field trial and large discrepancies of stimulated microorganisms in the laboratory and in the natural oil reservoir. More specifically, after nutrients injection, the original dominant populations of Petrobacter and Alishewanella in the production wells almost disappeared. The expected desirable population of Pseudomonas aeruginosa, determined by enrichment experiments in laboratory, was stimulated successfully in two wells of the five monitored wells. Unexpectedly, another potential population of Pseudomonas pseudoalcaligenes which were not detected in the enrichment culture in laboratory was stimulated in the other three monitored production wells. In this study, monitoring of microbial community displayed a comprehensive alteration of microbial populations during the field trial to remedy the deficiency of culture-dependent monitoring methods. The results would help to develop and apply more MEOR processes.     

Xanthan degragation by biofilm in porous media

Biological activity in oil reservoirs can cause significant problems such as souring and plugging. This study focuses on the problem of polymer degradation and permeability reduction due to biofilm formation during polymer injection for improved oil recovery. Polymers are included in injection fluids to increase their viscosity. Results relating biological processes and polymer degradation to fluid‐dynamic conditions in a laboratory model porous medium are presented.

A transparent flow cell with an etched two‐dimensional network of pores served as a model porous medium. A sterile xanthan polymer and natural sea water solution were continuously injected into the porous medium. A bacterial culture capable of xanthan degradation was introduced into the cell by a single injection. Some of the cells from this culture attached to the pore walls forming an immobile bacterial culture, termed biofilm. The development of this biofilm, its xanthan degradation and its effect on permeability were measured.

The effects of injection rate and rate transitions were analyzed. Injection fluid viscosity was reduced by 30% after 5 min flow through the porous medium at the maximum steady state degradation rate observed. Permeability was significantly reduced by the xanthan degrading biofilm, causing an increase in pressure drop through the porous medium of up to 80%. Polymer injection in oil reservoirs may, therefore, have negative effects on oil recovery, unless efficient biofouling control is applied. The methodology presented may serve as a tool in the development of biofouling control measures in porous media.     

Effect of exogenous inoculants on enhancing oil recovery and indigenous bacterial community dynamics in long-term field pilot of low permeability reservoir

Pseudomonas aeruginosa DN1 strain and Bacillus subtilis QHQ110 strain were chosen as rhamnolipid and lipopeptide producer respectively, to evaluate the efficiency of exogenous inoculants on enhancing oil recovery (EOR) and to explore the relationship between injected bacteria and indigenous bacterial community dynamics in long-term filed pilot of Hujianshan low permeability water-flooded reservoir for 26 months. Core-flooding tests showed that the oil displacement efficiency increased by 18.46% with addition of exogenous consortia. Bacterial community dynamics using quantitative PCR and high-throughput sequencing revealed that the exogenous inoculants survived and could live together with indigenous bacterial populations. They gradually became the dominant community after the initial activation, while their comparative advantage weakened continually after 3 months of the first injection. The bacterial populations did not exert an observable change in the process of the second injection of exogenous inoculants. On account of facilitating oil emulsification and accelerating bacterial growth with oil as the carbon source by the injection of exogenous consortia, γ-proteobacteria was finally the prominent bacterial community at class level varying from 25.55 to 32.67%, and the dominant bacterial populations were increased by 2–3 orders of magnitude during the whole processes. The content of organic acids and rhamnolipids in reservoir were promoted with the change of bacterial community diversity, respectively. Cumulative oil increments reached 26,190 barrels for 13 months after the first injection, and 55,947 barrels of oil had been accumulated in all of A20 wells block through two rounds of bacterial consortia injection. The performance of EOR has a cumulative improvement by the injection of exogenous inoculants without observable inhibitory effect on the indigenous bacterial populations, demonstrating the application potential in low permeability water-flooded reservoirs.     

Growth and Starvation of a Strain of Klebsiella pneumoniae Isolated from a Brazilian Oil Formation

Summary The changes that occur in cell size and shape during experimental starvation and resuscitation of bacteria may find practical application in the biotechnological processes used in microbial enhanced oil-recovery (MEOR), in in situ bioremediation and in the creation of biobarriers. In the work described here, the aim was to observe certain aspects of the response of a strain of Klebsiella pneumoniae, isolated from oil producer wells belonging to PETROBRAS (National Oil Company of Brazil), to starvation and growth under various culture conditions. Cells of K. pneumoniae were taken from an exponentially-growing culture, washed and suspended in phosphate-buffered saline solution, without nutrients, where they remained for 91 days. Aliquots were withdrawn periodically, to perform viable and total cell counts and measure the optical density and cell dimensions. The starved cells were resuspended in sodium citrate medium, where they recuperated and exhibited a typical growth curve. The results indicate that this strain is a viable option for future trials on the transport and growth of microorganisms inside porous media, aimed at possible applications in MEOR.     

培养法和免培养法联合检测油藏环境烃降解菌和产甲烷菌群多样性

烃降解菌和产甲烷菌是油藏环境微生物生态系统中重要的功能菌群, 采用DGGE和FISH方法分析了不同油藏样品中两类菌群的多样性和产甲烷活性。DGGE结果表明, 不同水样的alkB基因多样性相差较大, 而且注水井条带明显多于采油井。FISH结果表明, 油藏水样中产甲烷菌含量明显高于烃降解菌, 且两者空间分布的位置较近; 说明油藏环境中烃降解菌和产甲烷菌结成一定的相互关系。富集培养表明, 胜利油田产出液接种物培养130 d后, 石油烃降解率达到50%以上, 产甲烷的最大速率达到1.57×10^?2 mmol/(L?d)。利用分子生物学方法分析油藏环境功能菌群的多样性, 可以为开展微生物采油技术的应用提供有用信息。     

Application of microbial enhanced oil recovery technique to a Turkish heavy oil

Summary Microbial enhance oil recovery utilizes microorganisms and their metabolic products to improve the recovery of crude oil from reservoir rocks. In this study an anaerobic bacterium, Clostridium acetobutylicum was injected into a one-dimensional model reservoir containing a Turkish heavy oil (Raman oil) at 38° C. This injection was followed by water flooding after a suitable shut-in period. Comparison of oil recovery results of pure water flooding runs with experiments in which bacterial concentration and shut-in periods were varied indicated increases in oil recovery of about 12% of the original oil in place. This increase was attributed to changes in the viscosity and pH of the crude oil.Offprint requests to: T. Mehmetoglu     

Microbially enhanced oil recovery from carbonate reservoirs

About half of the world's oil production is from carbonate formations. However, most of the research in microbially enhanced oil recovery (MEOR), a potentially important tertiary recovery technology, has focused on sandstone reservoirs because, in general, they are geologically simpler than carbonate reservoirs and easier to model in the laboratory. Carbonate formations have a wide range of pore geometries and distributions, resulting in complex flow dynamics. The low matrix permeabilities and the dual porosity characteristics of most carbonate formations, coupled with the chemistry of carbonates, have slowed implementation of enhanced oil recovery methods. A review of the data on carbonate reservoirs in Dwight's Energydata TOTL System indicated that 40% of the oil‐producing carbonate reservoirs surveyed in the United States have environmental, geological, and petrophysical conditions that would make them candidates for MEOR. A review of a number of MEOR field trials showed that rates of oil production could be increased by as much as 200%. Microbial activity in these trials was probably due to that of indigenous populations rather than the microorganisms injected for the trials. Detrimental effects such as loss of injectivity and increased souring were not reported. Based on analysis of the geology and petrophysical characteristics of carbonates, two common mechanisms of MEOR, microbial acid production and microbial gas production, are especially suited for application in carbonate reservoirs.     

The potential for hydrocarbon biodegradation and production of extracellular polymeric substances by aerobic bacteria isolated from a Brazilian petroleum reservoir

Extracellular polymeric substances (EPS) can contribute to the cellular degradation of hydrocarbons and have a huge potential for application in biotechnological processes, such as bioremediation and microbial enhanced oil recovery (MEOR). Four bacterial strains from a Brazilian petroleum reservoir were investigated for EPS production, emulsification ability and biodegradation activity when hydrocarbons were supplied as substrates for microbial growth. Two strains of Bacillus species had the highest EPS production when phenanthrene and n-octadecane were offered as carbon sources, either individually or in a mixture. While Pseudomonas sp. and Dietzia sp., the other two evaluated strains, had the highest hydrocarbon biodegradation indices, EPS production was not detected. Low EPS production may not necessarily be indicative of an absence of emulsifier activity, as indicated by the results of a surface tension reduction assay and emulsification indices for the strain of Dietzia sp. The combined results gathered in this work suggest that a microbial consortium consisting of bacteria with interdependent metabolisms could thrive in petroleum reservoirs, thus overcoming the limitations imposed on each individual species by the harsh conditions found in such environments.     

Exopolysaccharide production by a genetically engineered Enterobacter cloacae strain for microbial enhanced oil recovery

Microbial enhanced oil recovery (MEOR) is a petroleum biotechnology for manipulating function and/or structure of microbial environments existing in oil reservoirs for prolonged exploitation of the largest source of energy. In this study, an Enterobacter cloacae which is capable of producing water-insoluble biopolymers at 37 °C and a thermophilic Geobacillus strain were used to construct an engineered strain for exopolysaccharide production at higher temperature. The resultant transformants, GW3-3.0, could produce exopolysaccharide up to 8.83 g l⁻¹ in molasses medium at 54 °C. This elevated temperature was within the same temperature range as that for many oil reservoirs. The transformants had stable genetic phenotype which was genetically fingerprinted by RAPD analysis. Core flooding experiments were carried out to ensure effective controlled profile for the simulation of oil recovery. The results have demonstrated that this approach has a promising application potential in MEOR.