Control of microbial sulfide production by limiting sulfate dispersal in a water-injected oil field.

Oil production by water injection often involves the use of makeup water to replace produced oil. Sulfate in makeup water is reduced by sulfate-reducing bacteria to sulfide, a process referred to as souring. In the MHGC field souring was caused by using makeup water with 4mM (384ppm) sulfate. Mixing with sulfate-free produced water gave injection water with 0.8mM sulfate. This was amended with nitrate to limit souring and was then distributed fieldwide. The start-up of an enhanced-oil-recovery pilot caused all sulfate-containing makeup water to be used for dissolution of polymer, which was then injected into a limited region of the field. Produced water from this pilot contained 10% of the injected sulfate concentration as sulfide, but was free of sulfate. Its use as makeup water in the main water plant of the field caused injection water sulfate to drop to zero. This in turn strongly decreased produced sulfide concentrations throughout the field and allowed a decreased injection of nitrate. The decreased injection of sulfate and nitrate caused major changes in the microbial community of produced waters. Limiting sulfate dispersal into a reservoir, which acts as a sulfate-removing biofilter, is thus a powerful method to decrease souring.

A screening test for the determination of ethylene sensitivity.

Ethephon, which releases ethylene within plant tissues after application, was chosen to perform assessments of the relative sensitivity of crops to ethylene and to determine which stages of plant development were most sensitive. The species chosen were: barley, wheat, oats, canola and field pea, all of which are important crops in the province of Alberta, Canada. Plants were treated with ethephon at one of 7 different stages. Plants were assessed for their vegetative and reproductive growth, including height, biomass, yield and seed quality. Visual symptoms were photographed and documented to compare them with symptoms caused by ethylene applied as a gas. It was concluded that in barley, wheat and canola the late vegetative and early reproductive stages were most sensitive, at least when sensitivity was defined as reductions in yield and quality. As for field pea, ethephon had no effect on yield but did cause increased numbers of pods, which in certain conditions could lead to increased yields. Significant effects on vegetative growth were only observed in the early vegetative stages of development but with no effects on yield. The screening protocol successfully identified sensitive cultivars and growth stages for further investigation of the effects of ethylene exposure.

Biodegradation of dicyclopentadiene in the field.

Dicyclopentadiene (DCPD) is formed during the pyrolysis of alkanes to produce olefins suitable for manufacturing synthetic polymers. DCPD has an irritating odor with a 5 ppb detection level that provides the impetus for remediation efforts. One method of destroying odors is to alter the structure of the chemical. This can be accomplished by biological oxidation using microorganisms. Field studies at two sites, where DCPD was a soil contaminant, indicated that biodegradation contributed significantly to DCPD removal. DCPD degradation was stimulated by decreasing bulk soil density and adding nitrogen and phosphorous nutrients. The presence of other easier degradable aromatic hydrocarbons may also be beneficial, suggesting that the process is cometabolic.

In vitro degradation of dicyclopentadiene by microbial consortia isolated from hydrocarbon-contaminated soil.

Degradation of dicyclopentadiene (DCPD) to carbon dioxide and oxygenated intermediates was established in the laboratory. Screening of many inocula using BIOLOGMT plates showed that no single colony isolate readily mineralized DCPD. Mixed cultures from a variety of environmental sources produced 14CO2 when incubated with [14C]DCPD, but most of the DCPD was metabolized to oxygenated intermediates that could be extracted from the culture liquid and detected using gas chromatography and mass spectroscopy. Stimulation of environmental inocula with nutrients and preexposure to DCPD before testing for degradation gave mineralization rates after 25 days of in vitro incubation that were twice as fast as those previously reported.

Disposal of slop oil and sludges by biodegradation.

These pilot tests indicate that immobilized microbial populations can degrade a wide range of crude oils absorbed into a properly prepared peat matrix with surprising speed and flexibility. Depending on the hydrocarbon composition, ultimate disposal by landfarming or landfilling of the residues is possible. In the former case, performance of the landfarm will be enhanced and environmental concerns related to its operation reduced. In terms of ease of operation, capital and operating costs and reduced environmental concerns, the novel bioreactor presented here meets the requirements and cost constraints associated with on-site slop oil and sludge disposal for the sorts of volumes normally encountered. A patent has been applied for (Francis and Jack, 1991). The technological issues in this development process largely arose from requirements and constraints associated with the target application. Scientific issues surrounding the enhanced biodegradation, seen especially for absorbed heavy oil, remain unresolved. These may or may not be picked up in further development and optimization as need and cost allow.

Quantitative reverse sample genome probing of microbial communities and its application to oil field production waters.

This paper presents a protocol for quantitative analysis of microbial communities by reverse sample genome probing is presented in which (i) whole community DNA is isolated and labeled in the presence of a known amount of an added internal standard and (ii) the resulting spiked reverse genome probe is hybridized with a master filter on which denatured genomic DNAs from bacterial standards isolated from the target environment were spotted in large amounts (up to 1,500 ng) in order to improve detection sensitivity. This protocol allowed reproducible fingerprinting of the microbial community in oil field production waters at 19 sites from which water and biofilm samples were collected. It appeared that selected sulfate-reducing bacteria were significantly enhanced in biofilms covering the metal surfaces in contact with the production waters.

Identification of distinct communities of sulfate-reducing bacteria in oil fields by reverse sample genome probing.

Thirty-five different standards of sulfate-reducing bacteria, identified by reverse sample genome probing and defined as bacteria with genomes showing little or no cross-hybridization, were in part characterized by Southern blotting, using 16S rRNA and hydrogenase gene probes. Samples from 56 sites in seven different western Canadian oil field locations were collected and enriched for sulfate-reducing bacteria by using different liquid media containing one of the following carbon sources: lactate, ethanol, benzoate, decanoate, propionate, or acetate. DNA was isolated from the enrichments and probed by reverse sample genome probing using master filters containing denatured chromosomal DNAs from the 35 sulfate-reducing bacterial standards. Statistical analysis of the microbial compositions at 44 of the 56 sites indicated the presence of two distinct communities of sulfate-reducing bacteria. The discriminating factor between the two communities was the salt concentration of the production waters, which were either fresh water or saline. Of 34 standards detected, 10 were unique to the fresh water and 18 were unique to the saline oil field environment, while only 6 organisms were cultured from both communities.

The surface active properties and antibacterial activity of the compounds, trans-[Rh(L)4Cl2]Cl . nH2O.

The antibacterial activity and surfactant activity of the compounds trans-[Rh(L)4-Cl2]Cl . nH2O increase in the order L = pyridine less than 4-methylpyridine less than 4-ethylpyridine less than 4-n-propylpyridine. As surfactants, the compounds are far more effective at reducing the interfacial tension, n-hexadecane/H2O, than the surface tension, H2O/air. The most effective and efficient surfactant in this series, trans-[Rh(4-n-propylpyridine)4Cl2]Cl . H2O, can cause the leakage of intracellular manganese ions from the gram-positive bacteria, Bacillus brevis ATCC 9999, at a concentration of 130 ppm but there is no observable effect on the retention of intracellular manganese ions at the minimum concentration required to prevent growth of this organism (approximately 0.6 ppm at 23 degrees C in nutrient broth). At 130 ppm, trans-[Rh(4-n-propylpyridine)4Cl2]Cl . H2O does not cause the loss of intracellular manganese ions from the gram-negative bacteria, Escherichia coli JS-1. In this case, a concentration of at least 63 ppm of this rhodium compound is required to prevent the growth of this organism in M9TUH medium at 35 degrees C. On the basis of these results, it is suggested that gross membrane disruption effects caused by the surfactants trans-[Rh(L)4Cl2A1Cl . nH2O are not directly responsible for their observed antibacterial action.

The effect of trans-[Rh(4-ethylpyridine)4Cl2]Cl x 2H2O on nucleic acid and protein syntheses in Escherichia coli.

The effect of the transition metal compound trans-[Rh(4-ethylpyridine)4Cl2]Cl x 2H2O on the syntheses of DNA, RNA, and protein has been investigated for an auxotrophic bacterial strain, Escherichia coli JS-1, incapable of thymidine, uridine, and histidine syntheses. At low concentration (7.4 x 10(-6) M), this rhodium complex interferes with normal cell division and induces the formation of filaments comparable to those observed in the presence of the cis-(NH3)2PtClx antitumour agents. Once the suppressed growth rate of the filamenting cells has been taken into account, the rhodium compound is found not to alter macromolecular synthesis. Again this is consistent with similar observations made for the platinum compounds.

The enzymatic conversion of L-histidine to urocanic acid by whole cells of Micrococcus luteus immobilized on carbodiimide activated carboxymethylcellulose.

Whole cells of Micrococcus luteus (formerly Sarcina lutea ATCC 9341) have been covalently linked to a carboxymethylcellulose support system, with the retention of histidine ammonia-lyase activity. The dependence of the rate of urocanic acid formation on pH, temperature, and added surfactant concentration was similar for the free and the immobilized cells. The immobilization procedure used is based on the carbodiimide activation of carboxymethylcellulose and has been optimized for the histidine ammonia-lyase activity of the immobilized cells on a given weight of cellulose. In a column reactor at 23 degrees C and superficial velocity of 0.044 cm/min, 5 g of cellulose with bound cells gave a 35% conversion of an L-histidine solution (0.25M, pH 9.0) to urocanic acid for 16 days of continuous operation. The scope of this carbodiimide assisted immobilization procedure has been investigated for a series of microorganisms and a variety of carboxylate functionalized supports.