V. S. Ivoilov

[Microbiological investigations of high-temperature horizons of the Kongdian petroleum reservoir in connection with field trial of a biotechnology for enhancement of oil recovery].

The physicochemical conditions and microbiological characteristics of the formation waters of the Kongdian oilfield of the Dagang oilfield (China) were studied. It was demonstrated that this oilfield is a high-temperature ecosystem with formation waters characterized by low mineralization. The concentrations of nitrogen and phosphorus compounds, as well as of electron acceptors, are low. Oil and oil gas are the main organic matter sources. The oilfield is exploited with water-flooding. The oil stratum was inhabited mostly by anaerobic thermophilic microorganisms, including fermentative (102–105 cells/ml), sulfate-reducing (0–102 cells/ml), and methanogenic (0–103 cells/ml) microorganisms. Aerobic bacteria were detected mainly in the near-bottom zone of injection wells. The rate of sulfate reduction varied from 0.002 to 18.940 μg S2− l−1 day−1 and the rate of methanogenesis from 0.012 to 16.235 μg CH4 l−1 day−1. Microorganisms with great biotechnological potential inhabited the oilfield. Aerobic thermophilic bacteria were capable of oxidizing oil with formation of biomass, the products of partial oxidation of oil (volatile acids), and surfactants. During growth on the culture liquid of oil-oxidizing bacteria, methanogenic communities produced methane and carbon dioxide, which also had oil-releasing capabilities. Using various labeled tracers, the primary filtration flows of injected solutions at the test site were studied. Our comprehensive investigations allowed us to conclude that the method for microbial enhancement of oil recovery based on the activation of the stratal microflora can be applied in the Kongdian oilfield.

Microbiological and production characteristics of the high-temperature Kongdian petroleum reservoir revealed during field trial of biotechnology for the enhancement of oil recovery

Microbiological technology for the enhancement of oil recovery based on the activation of the stratal microflora was tested in the high-temperature horizons of the Kongdian bed (60°C) of the Dagang oil-field (China). This biotechnology consists in the pumping of a water-air mixture and nitrogen and phosphorus mineral salts into the oil stratum through injection wells in order to stimulate the activity of the stratal microflora which produce oil-releasing metabolites. Monitoring of the physicochemical, microbiological, and production characteristics of the trial site has revealed large changes in the ecosystem as a result of the application of biotechnology. The cell numbers of thermophilic hydrocarbon-oxidizing, fermentative, sulfate-reducing, and methanogenic microorganisms increased 10–10000-fold. The rates of methanogenesis and sulfate reduction increased in the near-bottom zone of the injection wells and of some production wells. The microbial oil transformation was accompanied by the accumulation of bicarbonate ions, volatile fatty acids, and biosurfactants in the formation waters, as well as of CH4 and CO2 both in the gas phase and in the oil. Microbial metabolites promoted the additional recovery of oil. As a result of the application of biotechnology, the water content in the production liquid from the trial site decreased, and the oil content increased. This allowed the recovery of more than 14000 tons of additional oil over 3.5 years.

Regulation of geochemical activity of microorganisms in a petroleum reservoir by injection of H2O2 or water-air mixture

In the course of pilot trials of biotechnologies for the enhancement of oil recovery in formation waters of the Gangxi bed of the Dagang oil field (China), microbiological processes were investigated. The biotechnologies are based on injection into the petroleum reservoir of different oxygen sources (H2O2 solution or a water-air mixture) with nitrogen and phosphorus salts. The injection of water-air mixture with nitrogen and phosphorus salts resulted in an increase in the number of aerobic and anaerobic organotrophic bacteria, rates of sulfate reduction and methanogenesis in formation water and also the content of CO2 (from 4.8–12 to 15–23.2%) and methane (from 86–88 to 91.8%) in the gas. The preferential consumption of isotopically light bicarbonate by methanogens resulted in a higher content of the light 12C in methane; the δ13C/CH4 value changed from −45.1…−48.3 to −50.7…−59.3‰. At the same time, mineral carbonates of the formation water became isotopically heavier; the δ13C/Σcarbonates value increased from 3.4…4.0 to 5.4…9.6‰. Growth of hydrocarbon-oxidizing bacteria was accompanied by production of biosurfactants and decreased interfacial tension of formation water. Injection of H2O2 solution resulted in the activation of aerobic processes and in suppression of both sulfate reduction and methanogenesis. Methane content in the gas decreased from 86–88 to 75.7–79.8%, probably due to its consumption by methanotrophs. Due to consumption of isotopically light methane, the residual methane carbon became heavier, with the δ13C/CH4 values from −39.0 to −44.3‰. At the same time, mineral carbonates of the formation water became isotopically considerably lighter; the δ13C/Σcarbonates value decreased from 5.4…9.6 to −1.4…2.7‰. The additional amount of oil recovered during the trial of both variants of biotechnological treatment was 3819 t.

Degradation of Machine Oil by Nocardioform Bacteria

Gas liquid chromatography (GLC) was used for the first time to screen for machine oil–degrading microorganisms. Oil degradation was evaluated from the microorganism respiratory activity during the utilization of oil as the sole carbon and energy source. The results are consistent with those obtained by the conventional weighing method. Substrate specificity of the active strains with respect to different machine oils was studied. Bacterial communities exhibited the highest activity, whereas a Rhodococcus erythropolisstrain was the most active among pure cultures. Various stages of bacterial interaction with oil drops were followed by means of fluorescent microscopy.

Distribution and activity of microorganisms in the deep repository for liquid radioactive waste at the Siberian Chemical Combine

The physicochemical conditions, composition of microbial communities, and the rates of anaerobic processes in the deep sand horizons used as a repository for liquid radioactive wastes (LRW) at the Siberian Chemical Combine (Seversk, Tomsk oblast), were studied. Formation waters from the observation wells drilled into the horizons used for the radioactive waste disposal were found to be inhabited by microorganisms of different physiological groups, including aerobic organotrophs, anaerobic fermentative, denitrifying, sulfate-reducing, and methanogenic bacteria. The density of microbial population, as determined by cultural methods, was low and usually did not exceed 104 cells/ml. Enrichment cultures of microorganisms producing gases (hydrogen, methane, carbon dioxide, and hydrogen sulfide) and capable of participation in the precipitation of metal sulfides were obtained from the waters of the disposal site. The contemporary processes of sulfate reduction and methanogenesis were assayed; the rates of these terminal processes of organic matter destruction were found to be low. The denitrifying bacteria from the deep repository were capable of reducing the nitrates contained in the wastes, provided sources of energy and biogenic elements were available. Biosorption of radionuclides by the biomass of aerobic bacteria isolated from groundwater was demonstrated. The results obtained give us insight into the functional structure of the microbial community inhabiting the waters of repository horizons. This study indicates that the numbers and activity of microbial cells are low both inside and outside the zone of radioactive waste dispersion, in spite of the long period of waste discharge.

Microorganisms of the Carbonate Petroleum Reservoir 302 of the Romashkinskoe Oilfield and Their Biotechnological Potential

Diversity, geochemical activity, and biotechnological potential of the microorganisms from oil bed 302 of the Romashkinskoe oilfield (Tatarstan, Russia) are reported. The microbial community contained almost no aerobic microorganisms. Sulfate-reducing (103−106 cells/mL) and fermentative bacteria (102−105 cells/mL) predominated in the oilfield. Sulfate reduction was the predominant process in formation water with the rates up to 26.6 μg S2−L/day. The number of methanogens and methanogenesis rate in formation water did not exceed 104 cells/mL and 8.19 μg CH4 L/day, respectively. Analysis of the 16S rRNA gene clone library revealed the sequences of denitrifying bacteria of the genera Sulfurimonas and Thauera. The oil recovery technique combining the stimulation of fermentative bacteria and suppression of sulfate reducers in the oilfield was proposed for development of the bed 302. Fermentative bacteria could be activated by the traditional method, i.e., injection of molasses and nitrogen and phosphorus mineral salts through the injection wells. Introduction of high concentrations of nitrate will activate the growth of denitrifying bacteria, suppress the growth of sulfidogenic bacteria, and result in decreased sulfide concentration in formation water. The proposed biotechnology is technologically simple and environmentally friendly.

Microbiological processes in the Severnyi deep disposal site for liquid radioactive wastes

Local monitoring of physicochemical, radiochemical, and microbiological parameters was performed in the deep horizons of the Severnyi site used for disposal of liquid radioactive waste (LRW). Analysis of the chemical and radiochemical composition of the wastes and formation fluid revealed that the boundary for migration of radionuclides lagged behind that for nonradioactive waste components (sodium nitrate) and tritium. The physicochemical and radiochemical conditions in deep horizons did not prevent microbial growth. The numbers of microorganisms (aerobic organotrophs, denitrifying, fermentative, sulfate-reducing, and methanogenic) were low, as were the rates of sulfate reduction and methanogenesis; they increased in the waste dispersion zone. The microorganisms from deep horizons were able to produce gases (CH4, CO2, N2, and H2S) from possible waste components. Denitrifying bacteria belonged to different Pseudomonas species and reduced nitrate to dinitrogen under the conditions of pH, salinity, temperature, and radioactivity found in the disposal site. These results suggest the need for control of microbiological processes in deep disposal site for liquid RW.