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Radioisotopic, culture-based, and oligonucleotide microchip analyses of thermophilic microbial communities in a continental high-temperature petroleum reservoir.

Activity measurements by radioisotopic methods and cultural and molecular approaches were used in parallel to investigate the microbial biodiversity and its physiological potential in formation waters of the Samotlor high-temperature oil reservoir (Western Siberia, Russia). Sulfate reduction with rates not exceeding 20 nmol of H(2)S liter(-1) day(-1) occurred at 60 and 80 degrees C. In upper horizons (AB, A, and B), methanogenesis (lithotrophic and/or acetoclastic) was detected only in wells in which sulfate reduction did not occur. In some of the wells from deeper (J) horizons, high-temperature sulfate reduction and methanogenesis occurred simultaneously, the rate of lithotrophic methanogenesis exceeding 80 nmol of CH(4) liter(-1) day(-1). Enrichment cultures indicated the presence of diverse physiological groups representing aerobic and anaerobic thermophiles and hyperthermophiles; fermentative organotrophs were predominant. Phylogenetic analyses of 15 isolates identified representatives of the genera Thermotoga, Thermoanaerobacter, Geobacillus, Petrotoga, Thermosipho, and Thermococcus, the latter four being represented by new species. Except for Thermosipho, the isolates were members of genera recovered earlier from similar habitats. DNA obtained from three samples was hybridized with a set of oligonucleotide probes targeting selected microbial groups encompassing key genera of thermophilic bacteria and archaea. Oligonucleotide microchip analyses confirmed the cultural data but also revealed the presence of several groups of microorganisms that escaped cultivation, among them representatives of the Aquificales/Desulfurobacterium-Thermovibrio cluster and of the genera Desulfurococcus and Thermus, up to now unknown in this habitat. The unexpected presence of these organisms suggests that their distribution may be much wider than suspected.

Archaea↗

Mercury-resistant bacteria and petroleum degradation.

The concentration of mercury in water and sediment and in the oil extracted from water and sediment was determined for samples collected in Colgate Creek, located in Baltimore Harbor of the Chesapeake Bay. The concentration of mercury in the oil was 4,000 times higher than in sediment and 300,000 times higher than in water samples. The mercury-resistant bacterial populations of the samples studied have been shown to degrade oil, suggesting these bacteria to be a significant factor in the degradation of oil in Colgate Creek.

Bacteria↗

Degradation of petroleum by an alga, Prototheca zopfii.

Prototheca zopfii is an achlorophyllous alga which degrades oil. It has been found to degrade 10 and 40% of a motor oil and crude oil, respectively, when tested under appropriate conditions. Degradation of the crude oil observed in this study compares well with the amount of degradation accomplished by bacteria. P. zopfii was found to degrade a greater percentage of the aromatic hydrocarbons in motor oil than of the saturated hydrocarbons and a greater percentage of saturated hydrocarbons in crude oil than of aromatic hydrocarbons. Resins and asphaltens were produced during degradation of motor oil, whereas these fractions in crude oil were degraded. P. zopfii did not demonstrate preferential utilization of lower homologues of cycloalkanes and aromatics as has been observed with bacteria.

Alkanes↗

Effects of temperature and crude oil composition on petroleum biodegradation.

The biodegradability of seven different crude oils was found to be highly dependent on their composition and on incubation temperature. At 20 C lighter oils had greater abiotic losses and were more susceptible to biodegradation than heavier oils. These light crude oils, however, possessed toxic volatile components which evaporated only slowly and inhibited microbial degradation of these oils at 10 C. No volatile toxic fraction was associated with the heavier oils tested. Rates of oil mineralization for the heavier oils were significantly lower at 20 C than for the lighter ones. Similar relative degradation rates were found with a mixed microbial community, using CO2 evolution as the measure, and with a Pseudomonas isolate from the Arctic, using O2 consumption as the measure. The paraffinic, aromatic, and asphaltic fractions were subject to biodegradation. Some preference was shown for paraffin degradation, especially at low temperatures. Branched paraffins, such as pristane, were degraded at both 10 and 20 C. At best, a 20% residue still remained after 42 days of incubation. Oil residues generally had a lower relative percentage of paraffins and higher percentage of asphaltics than fresh or weathered oil.

Alkanes↗

An epidemiological study of petroleum refinery employees.

A cohort study of 14179 current and former Chevron USA employees at the Richmond and El Segundo, California, refineries was conducted. The cohort consisted of everyone working at either refinery for a minimum of one year. The observed mortality of the cohort, by cause, was compared with the expected based on the United States mortality rates, standardised for age, race, sex, and calendar time. Analyses by refinery, job category, hire date, duration of employment, and latency were performed. For the entire cohort, mortality from all causes was 72.4% of that expected, a deficit that was statistically significant. In addition, a significantly lower mortality was found for all forms of cancer combined, digestive cancer, lung cancer, heart disease, non-malignant respiratory disease, diseases of the digestive system, and accidents. Only lymphopoietic cancer showed a pattern of increased risk suggestive of a possible relation to an occupational exposure. The excess appears confined to cancer of lymphatic tissue (not leukaemias) at Richmond, and only among those hired before 1948. A follow up case analysis of the deaths from lymphatic cancer failed to identify a common exposure pattern.

Brain Neoplasms↗

An updated cause specific mortality study of petroleum refinery workers.

An update of a cohort study of 14,074 employees at the Richmond and El Segundo refineries of Chevron USA in California was conducted to further examine mortality patterns. The update added six years of follow up (1981-6) and 941 deaths. As in the previous study, mortality from all causes (standard mortality ratio (SMR) = 73) was significantly lower among men compared with the general United States population. Significant deficits were also found for all cancers combined (SMR = 81), several site specific cancers, and most non-malignant causes of death. Mortality from suicide was increased relative to the United States as a whole. Based on a comparison with California rates, however, men had fewer deaths from suicide than expected. Standard mortality ratios were raised for several other causes of death, but only leukaemia and lymphoreticulosarcoma exhibited a pattern suggestive of an occupational relation. The increase appeared to be confined to those hired before 1949, and in the case of lymphoreticulosarcoma, to Richmond workers.

California↗

A prospective study of morbidity patterns in a petroleum refinery and chemical plant.

This study examined the morbidity experience from 1981 to 1988 of a prospective cohort of 3422 refinery and petrochemical plant employees from the Shell Deer Park manufacturing complex. The morbidity data for this study, which include all illness and absence records in excess of five days, were extracted from the morbidity section of the Shell health surveillance system. Standardised morbidity ratios (SMRs) of disease prevalence in this cohort were calculated using an internal comparison group of all manufacturing employees of the Shell Oil Company. Among production employees, the overall morbidity was statistically significantly higher (SMR = 109) than that of the comparison group. Illness due to hypertension (SMR = 144), haemorrhoids (SMR = 149), diseases of the nervous system (SMR = 120), respiratory system (SMR = 108), and digestive system (SMR = 117) were also raised for this group. The increased risk due to these medical conditions does not appear to be associated with occupational factors. Lymphatic and haematopoietic tissue neoplasms were raised (SMR = 124), but were based on only four cases.

Acute Disease↗

Biodegradation of petroleum by Chesapeake Bay sediment bacteria.

Chesapeake Bay sediment bacteria from oil-contaminated and oil-free environments were compared for their ability to utilize a South Louisiana crude oil. Preferential solubility, column chromatography, gas-liquid chromatography, and computerized mass spectrometry were used to provide new and useful information regarding biodegradation of fractions and components of the crude oil. Vibrio, Pseudomonas, and Acinetobacter spp. were isolated from the culture inoculated with oil-contaminated sediment, whereas coryneforms and Pseudomonas spp. were isolated from the culture inoculated with oil-free sediment. Microorganisms from the oil-free sediment produced greater quantities of polar n-pentane-insoluble components (asphaltenes) after degradation, whereas microorganisms from the oil-contaminated sediments provided greater degradation of saturated and aromatic hydrocarbons.

Acinetobacter↗

Long-chain n-alkanes occurring during microbial degradation of petroleum.

Five axenic cultures and a mixed culture were examined for ability to degrade South Louisiana, Brass River Nigerian, Anaco Venezuelan, and Altamont crude oils. A wax was observed during microbial degradation of Altamont crude oil, but not during weathering of the oil. The high-boiling n-alkanes in the wax were associated with microbial degradation of the oil and appeared to be similar to components of tarballs found in the open ocean.

Acinetobacter↗

Biodegradation rates of components of petroleum.

Rates of microbial biodegradation of components of South Louisiana crude oil were determined by computer analysis of data obtained from computerized mass spectrometry. Total residue of the oil decreased exponentially with time, with maximum decrease noted at logarithmic phase, whereas asphaltenes and resins increased at the logarithmic phase of growth. Saturates decreased continuously during growth. Microbial degradation of components of crude oil was concluded to be a dynamic process.

Alkanes↗

Neural networks in petroleum engineering: a case study.

A multilayer neural network has been used for deciding which oil reservoir layer has to be perforated. Many network architectures were tested until we found those with the best generalization capability. The network performs better than human experts and its achievements are higher than the historical average in the test area. As in other applications of neural networks, the learning capability improves with more hidden neurons but the generalization does not.

Algorithms↗