Grantee Research Project Results
2002 Progress Report: Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
EPA Grant Number: R827015C021Subproject: this is subproject number 021 , established and managed by the Center Director under grant R827015
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: HSRC (1989) - Northeast HSRC
Center Director: Sidhu, Sukh S.
Title: Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
Investigators: McInerney, Michael , Suflita, Joseph , Gieg, Lisa M.
Institution: University of Oklahoma
EPA Project Officer: Aja, Hayley
Project Period: June 1, 2002 through May 31, 2004
Project Period Covered by this Report: June 1, 2002 through May 31, 2003
RFA: Integrated Petroleum Environmental Consortium (IPEC) (1999) RFA Text | Recipients Lists
Research Category: Hazardous Waste/Remediation , Targeted Research
Objective:
The objective of this research project is to determine the mechanism(s) of action of commercially available, microbial formulations used to treat paraffin deposition in the oil field. Because there are many conflicting reports by producers on the efficacy of microbial treatments to remedy paraffin deposits, it is not known why microbial treatments work under some conditions but not others. Knowledge of the mechanism(s) used by microorganisms to remediate paraffin deposits is a critical first step to understanding how the application of microbial treatments for paraffin removal can be optimized in the oil field. The results of this study will benefit the domestic oil industry because understanding the mechanisms of action of these products will allow the independent producer to determine the conditions under which they are likely to succeed and to determine if and when the purchase of microbial commercial paraffin treatments represents a wise expenditure of investment dollars.
Progress Summary:
We have completed the preliminary screening experiments for two crude oils (Alaska Oils A & B) selected by ConocoPhillips for this research project. These tests entailed incubating an oil with artificial brine medium in the absence (controls) or presence (tests) of a proprietary microbial product chosen specifically to treat the given oil. Tests were conducted at room temperature (~24 °C) and/or at 60 °C either in the presence or absence of oxygen. All incubations were in triplicate. Alaska Oil A was incubated for 3 days, whereas Alaska Oil B was incubated for 7 days (see below). After incubation with slow, end-over-end mixing, oil layers were removed to conduct the wax appearance temperature (WAT) test. Using cross-polarized microscopy, this test measures the temperature at which paraffin crystals begin to form when a given oil is cooled under controlled conditions. For the purposes of this research, microbial paraffin treatments that lowered the WAT by a minimum of 5 percent over that of the parallel microbial-free controls were considered successful for the preliminary screening assays. A lowering of the WAT values suggests that troublesome long-chain hydrocarbons have been “treated” by the microbial formulations.
For the preliminary screening of Alaska Oil A, all bottles were incubated for 3 days. Of the WAT tests conducted on Alaska Oil A, no bottles incubated under anaerobic conditions showed a reduction in the WAT. In contrast, one of the three replicates incubated aerobically at 60 °C showed an 8 percent reduction in the WAT relative to a microbial-free control. However, conducting ANOVA analysis of all of the WAT data from Alaska Oil A preliminary experiments showed that there was no significant difference in the WAT values for any microbe-amended bottle incubated under any condition relative to the microbe-free (sterile) controls. These results suggested that the proprietary microbial product selected to treat this oil was ineffective, at least over the 3-day incubation period. This outcome was somewhat surprising because previous data have shown that this oil was amenable to microbial treatment (ConocoPhillips, unpublished data). Further, emulsions were observed in many of the microbe-amended bottles but not in the sterile controls, suggesting microbial action on the oil. Three factors were postulated to have had an effect on the unsuccessful results: (1) cultures were incubated for too short of a time; (2) aerobic incubations became oxygen limited; and (3) WAT measurements for control incubations varied by as much as 10 percent, which made it difficult to determine a 5 percent difference from microbe-amended incubations.
Thus, to address the first two points for the preliminary experiments with Alaska Oil B, controls and cultures were incubated for 7 days instead of 3 days, and aerobic incubations were amended daily with sterile air to prevent oxygen deficiency. Further, WATs were conducted on controls at the onset of the analyses to evaluate the variability of oil samples removed from aqueous incubations. It was postulated that the wide differences in the WAT values observed in the control samples from Alaska Oil A may have been due to interferences from tiny emulsions with water droplets. For the controls in Alaska Oil B, at least duplicate measurements were made on the same drop of oil. The WAT values obtained from the controls incubated at 25°C (vary by ~28%) was much wider than that of the controls incubated at 60°C (vary by ~5%). Wax experts at ConocoPhillips suggested that the variability at 25°C is due to a phenomenon known as cold seeding in which paraffins drop out of the oil and move into an emulsion layer when the oil is below its cloud point, resulting in irreproducible oil samples. Thus, we concluded that the WAT test cannot accurately be performed on oils from room temperature incubations. Accordingly, incubations containing the microbial formulations were only established at 60°C. ANOVA tests performed on the WAT values measured at the end of the 7-day incubation period showed that there was a significant difference in the WAT value at 60°C under aerobic conditions relative to controls. We calculated a 7.2 percent reduction in the WAT value relative to controls. In contrast, under anaerobic conditions at 60°C, there were no statistically significant changes in the WAT values of the microbe-amended incubations relative to those of the controls (3.3% reduction in the WAT). The positive results observed at 60°C with Alaska Oil B have led us to establish Stage II experiments, which will help pinpoint the mechanism of action of the microbial formulation that caused the significant reduction in the WAT values. These experiments are currently incubating. In this phase of the project, analytical methods for examining oils have been developed and will be used to determine whether changes in the long-chain hydrocarbons are evident as a result of microbial treatment at the end of Stage II experiments. High-temperature gas chromatography methods (e.g., simulated distillation) have been developed to analyze standard mixtures of long-chain hydrocarbons and paraffin-rich oils. Our analytical studies have shown that the proportion of the higher molecular waxes is small relative to the other hydrocarbons in the oil, and it will be difficult to quantitate their loss. Thus, we will have to perform a wax extraction procedure to concentrate these waxes for quantitative analysis. Currently, this procedure is being developed and will follow the method of Thanh, et al. (1999). Stage II experiments also will involve surface tension measurements to determine whether biosurfactant production is a mechanism used by the microbes in the formulation to help treat or reduce paraffin accumulation. This measurement uses a DuNuoy ring detachment apparatus, which has been calibrated using water and isopropanol as high- and low-end standards, respectively. Further, an emulsification assay (Trebbau, et al., 1996) will be conducted to determine whether the microbes in the formulation are acting to treat paraffin by secreting emulsifying agents.
Future Activities:
At the end of a long incubation period (40 to 60 days), Stage II experiments with Alaska Oil B will be assayed for biosurfactant- or emulsifier-producing activities, and for changes in the hydrocarbon profiles relative to sterile controls. Further, supernatants of the incubations also will be analyzed for the presence of known hydrocarbon biodegradation products to determine whether this is a mechanism used by microbes to treat paraffin-rich oil. Also, we are hoping to identify a third oil on which to conduct microbial paraffin-treatment studies.
References:
de Acevedo GT, McInerney MJ. Emulsifying activity in thermophilic and extremely thermophilic microorganisms. Journal of Industrial Microbiology 1996;16(1):1-7.
Thanh NX, Hsieh M, Philp RP. Waxes and asphaltenes in crude oils. Organic Geochemistry 1999;30(2-3):119-132.
Journal Articles:
No journal articles submitted with this report: View all 1 publications for this subprojectSupplemental Keywords:
paraffin, microbial treatment, petroleum, bioremediation, microbiology,, RFA, Scientific Discipline, INTERNATIONAL COOPERATION, TREATMENT/CONTROL, Waste, Sustainable Industry/Business, Sustainable Environment, Environmental Chemistry, Technology, Analytical Chemistry, Technology for Sustainable Environment, Economics and Business, Ecological Risk Assessment, Technical Assistance, Ecology and Ecosystems, Bioremediation, pollution prevention, Environmental Engineering, chemical waste, clean technologies, cleaner production, microbial degradation, hazardous emissions, petrochemicals, oil production, biodegradation, oil production tank bottoms, hazardous waste, pollution control, IPEC, anaerobic biodegradation, paraffin deposition, innovative technology, technology transfer, technology researchRelevant Websites:
Progress and Final Reports:
Original AbstractMain Center Abstract and Reports:
R827015 HSRC (1989) - Northeast HSRC Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R827015C001 Evaluation of Road Base Material Derived from Tank Bottom Sludges
R827015C002 Passive Sampling Devices (PSDs) for Bioavailability Screening of Soils Containing Petrochemicals
R827015C003 Demonstration of a Subsurface Drainage System for the Remediation of Brine-Impacted Soil
R827015C004 Anaerobic Intrinsic Bioremediation of Whole Gasoline
R827015C005 Microflora Involved in Phytoremediation of Polyaromatic Hydrocarbons
R827015C006 Microbial Treatment of Naturally Occurring Radioactive Material (NORM)
R827015C007 Using Plants to Remediate Petroleum-Contaminated Soil
R827015C008 The Use of Nitrate for the Control of Sulfide Formation in Oklahoma Oil Fields
R827015C009 Surfactant-Enhanced Treatment of Oil-Contaminated Soils and Oil-Based Drill Cuttings
R827015C010 Novel Materials for Facile Separation of Petroleum Products from Aqueous Mixtures Via Magnetic Filtration
R827015C011 Development of Relevant Ecological Screening Criteria (RESC) for Petroleum Hydrocarbon-Contaminated Exploration and Production Sites
R827015C012 Humate-Induced Remediation of Petroleum Contaminated Surface Soils
R827015C013 New Process for Plugging Abandoned Wells
R827015C014 Enhancement of Microbial Sulfate Reduction for the Remediation of Hydrocarbon Contaminated Aquifers - A Laboratory and Field Scale Demonstration
R827015C015 Locating Oil-Water Interfaces in Process Vessels
R827015C016 Remediation of Brine Spills with Hay
R827015C017 Continuation of an Investigation into the Anaerobic Intrinsic Bioremediation of Whole Gasoline
R827015C018 Using Plants to Remediate Petroleum-Contaminated Soil
R827015C019 Biodegradation of Petroleum Hydrocarbons in Salt-Impacted Soil by Native Halophiles or Halotolerants and Strategies for Enhanced Degradation
R827015C020 Anaerobic Intrinsic Bioremediation of MTBE
R827015C021 Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
R827015C022 A Continuation: Humate-Induced Remediation of Petroleum Contaminated Surface Soils
R827015C023 Data for Design of Vapor Recovery Units for Crude Oil Stock Tank Emissions
R827015C024 Development of an Environmentally Friendly and Economical Process for Plugging Abandoned Wells
R827015C025 A Continuation of Remediation of Brine Spills with Hay
R827015C026 Identifying the Signature of the Natural Attenuation of MTBE in Goundwater Using Molecular Methods and "Bug Traps"
R827015C027 Identifying the Signature of Natural Attenuation in the Microbial
Ecology of Hydrocarbon Contaminated Groundwater Using Molecular Methods and
"Bug Traps"
R827015C028 Using Plants to Remediate Petroleum-Contaminated Soil: Project Continuation
R827015C030 Effective Stormwater and Sediment Control During Pipeline Construction Using a New Filter Fence Concept
R827015C031 Evaluation of Sub-micellar Synthetic Surfactants versus Biosurfactants for Enhanced LNAPL Recovery
R827015C032 Utilization of the Carbon and Hydrogen Isotopic Composition of Individual Compounds in Refined Hydrocarbon Products To Monitor Their Fate in the Environment
R830633 Integrated Petroleum Environmental Consortium (IPEC)
R830633C001 Development of an Environmentally Friendly and Economical Process for Plugging Abandoned Wells (Phase II)
R830633C002 A Continuation of Remediation of Brine Spills with Hay
R830633C003 Effective Stormwater and Sediment Control During Pipeline Construction Using a New Filter Fence Concept
R830633C004 Evaluation of Sub-micellar Synthetic Surfactants versus Biosurfactants for Enhanced LNAPL Recovery
R830633C005 Utilization of the Carbon and Hydrogen Isotopic Composition of Individual Compounds in Refined Hydrocarbon Products To Monitor Their Fate in the Environment
R830633C006 Evaluation of Commercial, Microbial-Based Products to Treat Paraffin Deposition in Tank Bottoms and Oil Production Equipment
R830633C007 Identifying the Signature of the Natural Attenuation in the Microbial Ecology of Hydrocarbon Contaminated Groundwater Using Molecular Methods and “Bug Traps”
R830633C008 Using Plants to Remediate Petroleum-Contaminated Soil: Project Continuation
R830633C009 Use of Earthworms to Accelerate the Restoration of Oil and Brine Impacted Sites
The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.
Project Research Results
Main Center: R827015
120 publications for this center
16 journal articles for this center