Lewis R. Brown

Microbial enhanced oil recovery (MEOR)

Two-thirds of the oil ever found is still in the ground even after primary and secondary production. Microbial enhanced oil recovery (MEOR) is one of the tertiary methods purported to increase oil recovery. Since 1946 more than 400 patents on MEOR have been issued, but none has gained acceptance by the oil industry. Most of the literature on MEOR is from laboratory experiments or from field trials of insufficient duration or that lack convincing proof of the process. Several authors have made recommendations required to establish MEOR as a viable method to enhance oil recovery, and until these tests are performed, MEOR will remain an unproven concept rather than a highly desirable reality.

Effects of SEM Preparation Techniques on the Appearance of Bacteria and Biofilms in the Carter Sandstone

Abstract When biofilms (aggregations of bacteria and extracellular polymer secretions) in samples from the Carter Sandstone of Alabama were prepared for scanning electron microscopy (SEM) using different dehydration techniques, the organic material had visibly different textures and distributions. In order to assess whether the variation was attributable to sample preparation or to inherent biofilm heterogeneity, each of five techniques were tested 3 to 10 times on small (1 cm) pieces of the Carter Sandstone containing either a strain of bacteria cultured from and reintroduced into the rock, or an in situ biofilm grown by injection of nutrients through core samples. The techniques tested were (1) air drying alone; (2) fixation in 10% glutaraldehyde with air drying; (3) ethanol dehydration with hexamethyldisilazane (HMDS) drying [2.5% glutaraldehyde, ethanol dehydration, and HMDS]; (4) ethanol dehydration with critical-point drying; and (5) ethanol and acetone dehydration with critical-point drying. Unpreserved control samples were either imaged wet in an environmental scanning electron microscope (ESEM) or vacuum-dried for SEM. Observations were based on SEM microscopy of over 60 samples and study of over 150 photomicrographs. In both experiments, the original morphology of individual bacteria was best preserved by ethanol dehydration with HMDS drying, ethanol dehydration with critical-point drying, or ethanol-acetone dehydration with critical-point drying. Critical-point drying preserved bacteria but stripped away mucilaginous material, revealing filamentous structures within the biofilm. These filaments, along with masses of microspheres (nannobacteria?) and the smooth mucilaginous outer layer, also occur in wet samples studied by ESEM, and are, therefore, not dehydration artifacts. However, different sample preparation techniques accentuated different components of the heterogeneous biofilm, thus resulting in vastly different textures. The cultured bacteria produced a biofilm that had a different surface texture and was more susceptible to sample preparation artifacts than the in situ biofilm. Use of more than one sample preparation technique is recommended in order to avoid bias.

Methanogenesis under acidic pH conditions in a semi-continuous reactor system

Operating an anaerobic digester at low pH could offer several advantages over operation at neutral pH. Most wastewater streams targeted for anaerobic digestion are inherently acidic, requiring alkalinity supplementation (at added expense) to buffer the pH at neutral. Additionally, previously published work completed by the authors using batch systems suggested that lowering the system pH could increase methane production by as much as 30%. The goal of this research was to evaluate the feasibility of sustaining methanogenesis at low pH in a semi-continuous laboratory-scale fermentor. Significant methane production was achieved in a system ranging in pH from approximately 4.0-5.3. Results show that, if the consortium is allowed to sufficiently acclimate to acidic conditions, methanogenesis can be maintained under acidic pH conditions, resulting in overall chemical oxygen demand (COD) reduction and methane production comparable to that achieved in a neutral pH system.

Use of Sublimation To Prepare Solid Microbial Media with Water-Insoluble Substrates

ABSTRACT A method was developed to deposit a visible layer of water-insoluble compounds via sublimation onto the surface of solid media. The compound is sublimed from a heated aluminum dish containing the compound onto the surface of an inverted, ice-cooled, inoculated agar petri dish. The method results in the deposition of a thin, even layer on the agar surface without the use of solvent. After incubation, clearing zones around colonies indicate the presence of compound-degrading microorganisms.

Examination of thirteen petroliferous formations for hydrocarbon-utilizing sulfate-reducing microorganisms

Virgin cores and production fluids were obtained from seven wells, ranging in depth from 805 ft to 14 492 ft, and examined for the presence of sulfate-reducing bacteria (SRB) using Rosenfelds sulfate-reducing medium modified by using crude oil in place of lactate. Cores from an additional six wells, ranging in depth from 1160 ft to 13 337 ft were tested for SRB using the modified Rosenfeld medium and API-sulfate-reducing medium. Produced waters from five of the six wells were tested also. All of the eleven produced water samples were positive for SRB while H2S production was not detected from the core samples.

Isopropanol and acetone induces vinyl chloride degradation in Rhodococcus rhodochrous

Abstract In situ bioremediation of vinyl chloride (VC)-contaminated waste sites requires a microorganism capable of degrading VC. While propane will induce an oxygenase to accomplish this goal, its use as a primary substrate in bioremediation is complicated by its flammability and low water solubility. This study demonstrates that two degradation products of propane, isoproponal and acetone, can induce the enzymes in Rhodococcus rhodochrous that degrade VC. Additionally, a reasonable number of cells for bioremediation can be grown on conventional solid bacteriological media (nutrient agar, tryptic soy agar, plate count agar) in an average microbiological laboratory and then induced to produce the necessary enzymes by incubation of a resting cell suspension with isopropanol or acetone. Since acetone is more volatile than isopropanol and has other undesirable characteristics, isopropanol is the inducer of choice. It offers a non-toxic, water-soluble, relatively inexpensive alternative to propane for in situ bioremediation of waste sites contaminated with VC.

The Utilization of the Microflora Indigenous to and Present in Oil-Bearing Formations to Selectively Plug the More Porous Zones Thereby Increasing Oil Recovery During Waterflooding

This project was designed to demonstrate that a microbially enhanced oil recovery process (MEOR), developed in part under DOE Contract No. DE-AC22-90BC14665, will increase oil recovery from fluvial dominated deltaic oil reservoirs. The process involves stimulating the in-situ indigenous microbial population in the reservoir to grow in the more permeable zones, thus diverting flow to other areas of the reservoir, thereby increasing the effectiveness of the waterflood. This five and a half year project is divided into three phases, Phase I, Planning and Analysis (9 months), Phase II, Implementation (45 months), and Phase III, Technology Transfer (12 months). Phase I was completed and reported in the first annual report. This fifth annual report covers the completion of Phase II and the first six months of Phase III.

Improvement of Carbon Dioxide Sweep Efficiency by Utilization of Microbial Permeability Profile Modification to Reduce the Amount of Oil Bypassed During Carbon Dioxide Flood

The objective of this project was to couple microbial permeability profile modification (MPPM), with carbon dioxide flooding to improve oil recovery from the Upper Cretaceous Little Creek Oil Field situated in Lincoln and Pike counties, MS. This study determined that MPPM technology, which improves production by utilizing environmentally friendly nutrient solutions to simulate the growth of the indigenous microflora in the most permeable zones of the reservoir thus diverting production to less permeable, previously unswept zones, increased oil production without interfering with the carbon dioxide flooding operation. Laboratory tests determined that no microorganisms were produced in formation waters, but were present in cores. Perhaps the single most significant contribution of this study is the demonstration that microorganisms are active at a formation temperature of 115⁰C (239⁰F) by using a specially designed culturing device. Laboratory tests were employed to simulate the MPPM process by demonstrating that microorganisms could be activated with the resulting production of oil in coreflood tests performed in the presence of carbon dioxide at 66˚C (the highest temperature that could be employed in the coreflood facility). Geological assessment determined significant heterogeneity in the Eutaw Formation, and documented relatively thin, variably-lithified, well-laminated sandstone interbedded with heavily-bioturbated, clay-rich sandstone and shale. Live core samples of the Upper Cretaceous Eutaw Formation from the Heidelberg Field, MS were quantitatively assessed using SEM, and showed that during MPPM permeability modification occurs ubiquitously within pore and throat spaces of 10-20 μm diameter. Testing of the MPPM procedure in the Little Creek Field showed a significant increase in production occurred in two of the five production test wells; furthermore, the decline curve in each of the production wells became noticeably less steep. This project greatly extends the number of oil fields in which MPPM can be implemented.

Effects of n-hexadecane and PM-100 clay on trichloroethylene degradation by Burkholderia cepacia

Trichloroethylene (TCE) is a non-flammable, volatile organochlorine compound which was a widely used degreasing agent, anesthetic, and coolant prior to 1960, but has since been placed on the Environmental Protection Agencys (EPA) list of priority pollutants. The inadequate disposal practices for TCE have created numerous TCE-contaminated superfund sites. The most commonly employed practice for remediating TCE-contaminated sites is to purge the contaminant from the source and trap it onto an adsorbent which is disposed of in a landfill or by incineration. This investigation was undertaken to evaluate the effectiveness of Burkholderia cepacia strain G4 (G4) to regenerate used sorbents by degrading TCE from the sorbent directly or indirectly. The results of this investigation showed that G4 was capable of reducing TCE attached to PM-100 clay but at significantly reduced rate due to the slow desorption rate. Conversely, it was shown that G4 was capable of degrading TCE dissolved in n-hexadecane at the same rate as systems without n-hexadecane present. The reduction in TCE degradation when the TCE is attached to the PM-100 clay could be overcome by solvent rinsing the TCE from the clay with subsequent removal of the TCE from the n-hexadecane by G4.

An Oligochaete as a Potential Food Source for Fish in Aquaculture

Abstract The oligochaete Aeolosoma heinprichi was successfully cultured in an aquatic environment with single-cell protein as its sole food source. Aeolosoma heinprichi grew throughout a pH range of 5.0–7.0 and a temperature range of 20–32°C. Decreased pH caused an increased doubling time (DT), i.e., a decreased reproductive rate. Increased temperatures resulted in decreased DT; growth was similar between 25 and 32°C. At pH 7.0 and 25°C, minimum DT was 36 h. Population densities depended on the bottom area of the culture vessel and reached a maximum of 330 organisms/cm2. Moderate concentrations of inorganic ions were toxic to the organism. The oligochaete proved to be an adequate food for fry and adult guppies (Poecilia reticulata) and adult golden shiners (Notemigonus crysoleucas).