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Determination of chromosome aberrations in workers in a petroleum refining factory.

Human exposure to benzene is derived occupationally from the petrochemical and petroleum refining industries. This study was performed to determine whether the frequencies of chromosome aberrations in workers exposed to low levels of benzene in a petroleum refining factory were elevated compared to an unexposed control group. The study population was comprised of 178 exposed workers and 36 unexposed workers. The frequencies of chromatid deletions and total chromosome aberrations in workers exposed to benzene were significantly higher compared to the unexposed control group. The frequency of total chromosome aberration was 4.20 per 500 metaphase cells in the exposed workers, whereas the frequency was 2.24 per 500 metaphase cells in the unexposed control group. The frequencies of total chromosome aberrations were significantly associated with benzene concentration after adjusting for confounding variables such as age, smoking status, and alcohol intake. The frequencies of chromosome aberrations were significantly increased in workers with low white blood cell counts (below 4000 cells/mm3) compared to those with high white blood cell counts (above 4000 cells/mm3). A reduced white blood cell count (below 4000/mm3) is suggestive of chronic exposure to benzene. In conclusion chronic benzene exposure and preclinical signs, such as reduced white blood cell counts, may be monitored by chromosome aberrations analysis.

Adult↗

Chemical composition and genotoxic activity of petroleum derivatives collected in two working environments.

Pitch and bitumen, two complex petroleum derivative mixtures, were studied for both their chemical composition and their mutagenic/DNA damaging activity. While bitumen revealed no genotoxic effect and low polycyclic aromatic hydrocarbons (PAHs) concentration, petroleum pitch showed a high concentration of mutagenic/carcinogenic PAHs, and also an elevated mutagenic activity when assayed by the Ames test, in the presence of postmitochondrial rat liver fractions. The in vitro mutagenic activity was detectable as frameshift mutation by assaying the pitch both as an in toto mixture and after HPLC fractionation, the most polar fractions being the most active. In contrast, both derivatives showed no in vivo DNA damage in rat liver, using the DNA alkaline elution technique and the fluorometric assay of DNA unwinding.

Animals↗

Benzene and total hydrocarbons exposures in the downstream petroleum industries.

A review of studies, including both articles published in peer-reviewed journals and reports that were not peer reviewed, regarding occupational exposure to benzene and total hydrocarbons in the downstream petroleum industry operations was performed. The objective was to provide a broad estimate of exposures by compiling exposure data according to the following categories: refinery, pipeline, marine, rail, bulk terminals and trucks, service stations, underground storage tanks, tank cleaning, and site remediations. The data in each category was divided into personal occupational long-term and short-term samples. The summarized data offers valuable assistance to hygienists by providing them with an estimate and range of exposures. The traditional 8-hour time-weighted average (TWA) exposure and the 40-hour workweek do not generally coincide with exposure periods applicable to workers in marine, pipeline, railcar, and trucking operations. They are more comparable with short-term exposure or task-based exposure assessments. The marine sector has a large number of high exposures. Although relatively few workers are exposed, their exposures to benzene and total hydrocarbons are sometimes an order of magnitude higher than the respective exposure limits. It is recommended that in the future, it would be preferable to do more task-based exposure assessments and fewer traditional TWA long-term exposure assessments within the various sectors of the downstream petroleum industry.

Air Pollutants, Occupational↗

Hydrocarbon exposures at petroleum bulk terminals and agencies.

Occupational exposures to the 55 hydrocarbon components of gasoline and petroleum products were measured at the bulk terminals and agencies of six Ontario petroleum companies during the summer of 1986. A total of 82 long-term (full-shift) and 111 short-term personal samples were taken over 3 months. The data, expressed as concentrations in milligrams per cubic meter, were highly variable and appeared to fit the lognormal distribution well. Full-shift exposures of bulk terminal drivers, agency drivers, and plantmen to total hydrocarbons (THC), computed as an n-hexane equivalent, and other hydrocarbon components for which exposure limits exist can be expected to exceed their respective 1986-1987 threshold limit value-time-weighted average (TLV-TWA) no greater than 1% of the time on the basis of the lognormal model. The short-term THC exposures of agency truck drivers can be expected to exceed the 1986-1987 TLV-short-term exposure limits about 7% of the time while top-loading and more than 17% while off-loading. For benzene, the short-term exceedance percentages are 1% and 4% for top- and off-loading operations, respectively. For long-term benzene exposures, up to 69% of the assessments can be expected to exceed the 1990-1991 proposed TLV-TWA of 0.3 mg/m3 (0.1 ppm). The full-shift hydrocarbon exposures of agency drivers were significantly higher than those for bulk terminal drivers. At the bulk terminals, the short-term hydrocarbon exposures during top-loading were significantly higher than during bottom-loading.

Dose-Response Relationship, Drug↗

A mathematical model of phytoremediation for petroleum contaminated soil: sensitivity analysis.

Phytoremediation is an attractive treatment technology for many contaminated sites due to its cost effectiveness and public acceptance. We present a sensitivity analysis of important parameters from a screening level model for phytoremediation by grass species of weathered petroleum-contaminated sites. The conceptual framework is that root movement through contaminated soil will enhance contaminant biodegradation by providing a local environment more favorable for petroleum degrading microorganisms--the so-called rhizosphere effect. Common questions in phytoremediation are, "What species should be planted?" and "What management practices should be followed?" These choices may affect degradation kinetics, root biomass (and therefore rhizosphere volume), and the root turnover. Important model parameters are the rate constants, rhizosphere volume, and the rate of root turnover. We present a sensitivity analysis with the aim of identifying the most important factors for improving phytoremediation effectiveness. For simulations of the phytoremediation of weathered diesel range organics, our results indicate that annual species, with higher root turnover, are preferred over perennial species with the caveat of equal degradation rate constants, that is, no species-dependent effects. In addition, the results suggest that the management of nonrhizosphere soil could play an important role in the overall effectiveness of phytoremediation. Finally, the effect of increasing root biomass or increasing the rhizosphere thickness is approximately equivalent with respect to the ultimate removal of the contaminants.

Environmental Pollution↗

Application of glass capillary columns to monitor petroleum-type hydrocarbons in marine sediments.

High resolution glass capillary columns coated with SE 52 liquid phase were used to resolve the indigenous hydrocarbons extracted from sediment samples collected from three outer continental shelf areas. The extracts were than spiked with small amounts of aliphatic or aromatic components isolated from a Louisiana crude oil and rechromatographed. The resolution was sufficient to separate almost all the isoprenoid, branched and cyclic alkanes associated with petroleum. Additionally, many of the key petroleum aromatics could also be resolved from naturally occurring organics. Such chromatographic procedures will aid in distinguishing between indigenous hydrocarbons of contemporary origin and those known to be associated with fossil hydrocarbon pollution of marine sediments.

Chromatography, Gas↗

Toxicology of petroleum naphtha distillate vapors.

A unique opportunity was presented to observe the potentially toxic effects of an acute exposure to the vapors of petroleum naphtha distillate on a relatively large number of individuals. The immediate manifestation in all was dyspnea. The action on motor vehicle combustion suggested that some of this could have been due to oxygen deprivation; however, all individuals were dyspneic for several minutes after exposure. A few were cyanotic for several minutes after exposure. All were excited. Tremulousness and mild nausea followed the initial symptoms but were of brief duration. One individual manifested numerous premature ventricular contractions. Since his exposure was brief and since none of the others showed similar findings, it is unlikely that the exposure was causal. The central nervous system depression described in acute exposure cases of the intact (not distillate) petroleum naphtha fumes was not observed in any of this series. There were no delayed manifestations or complications.

Alkanes↗

Radiological assessment of petroleum pipe scale from pipe-rattling operations.

Petroleum pipe scale, consisting of concentrated inorganic solids such as barium sulfate, can deposit on the inside of down-hole pipes during the normal course of oil field pumping operations. A portion of this scale has been shown to contain naturally occurring radioactive materials (NORM), predominantly compounds of radium. When these pipes are removed from the well, there is a potential for radiation doses to the oil field workers handling the pipes, especially as the pipes are cleaned for reuse. A thorough sampling and measurement protocol was applied under a variety of weather conditions in an outdoor laboratory to obtain an accurate indication of the radiological and aerodynamic characteristics of scale release and dust dispersion during petroleum pipe scale removal from out-of-service pipes with a restored, historically relevant outdoor pipe-cleaning machine. Exposure rate data were also obtained for both the pre-cleaned pipes, and the general area inhabited by workers during the descaling operation. Four radiation exposure pathways were investigated: inhalation of pipe scale dust generated during pipe rattling, incidental ingestion of the pipe scale dust, external exposure from uncleaned pipes, and external exposure from pipe scale dispersed on the ground. Pipes from three oil fields were rattled to collect as much industry-representative data as possible. The Ra specific activity of the pipe scale ranged from 33.6 +/- 0.4 to 65.5 +/- 0.7 Bq g, depending on the formation. A median atmospheric dust loading of 0.13 mg m was measured in the operator breathing zone. The respirable fraction was observed to be about 42% to 46%. Based on cleaning 20 pipes per day,250 d per year on average, annual committed effective doses for the operator and helper ranged from 0.11 mSv (11 mrem) to 0.45 mSv(45 mrem) for inhalation and from 19 microSv (1.9 mrem) to 97 microSv (9.7 mrem) for incidental ingestion. Worker annual external dose from the pipe racks ranged from 0 to 0.28 mSv (28 mrem). In the deposition experiment, more than 99% by weight of the deposited scale fell within 2 m of the machine centerline, the vast majority of which was in the downwind direction. The dose from this deposited material dominated the worker dose estimates. The annual external dose from dispersed material was estimated to be 2.8 mSv (280 mrem) for the operator and 4.1 mSv (410 mrem) for the helper.

Barium Sulfate↗

Physiological and molecular characterization of a microbial community established in unsaturated, petroleum-contaminated soil.

The microbial communities established in soil samples from an unsaturated, petroleum-contaminated zone and from an adjacent uncontaminated site were characterized by physiological and molecular approaches. Possible electron acceptors such as sulfate and nitrate had been completely depleted in these soil samples. Slurries of these soil samples were incubated in bottles in the presence of hydrocarbon indicators (benzene, toluene, xylene and decane), and the degradation of these compounds was examined. Supplementation with electron acceptors stimulated hydrocarbon degradation, although the stimulatory effect was small in the contaminated soil. The initial degradation rates in the contaminated soil under fermentative/methanogenic conditions were comparable to those under aerobic conditions. The microbial populations in the original soil samples were analysed by cloning and sequencing of polymerase chain reaction (PCR)-amplified bacterial and archaeal 16S rRNA gene fragments, showing that the sequences retrieved from these soils were substantially different. For instance, Epsilonproteobacteria, Gammaproteobacteria, Crenarchaeota and Methanosarcinales could only be detected at significant levels in the contaminated soil. Denaturing gradient gel electrophoresis (DGGE) analyses of 16S rRNA gene fragments amplified by PCR from the incubated soil-slurry samples showed that supplementation of the electron acceptors resulted in a shift in the major populations, while the DGGE profiles after incubating the contaminated soil under the fermentative/methanogenic conditions were not substantially changed. These results suggest that petroleum contamination of the unsaturated zone resulted in the establishment of a fermentative/methanogenic community with substantial hydrocarbon-degrading potential.

Bacteria↗

Rapid evolution of redox processes in a petroleum hydrocarbon-contaminated aquifer.

Ground water chemistry data collected over a six-year period show that the distribution of contaminants and redox processes in a shallow petroleum hydrocarbon-contaminated aquifer has changed rapidly over time. Shortly after a gasoline release occurred in 1990, high concentrations of benzene were present near the contaminant source area. In this contaminated zone, dissolved oxygen in ground water was depleted, and by 1994 Fe(III) reduction and sulfate reduction were the predominant terminal electron accepting processes. Significantly, dissolved methane was below measurable levels in 1994, indicating the absence of significant methanogenesis. By 1996, however, depletion of solid-phase Fe(III)-oxyhydrox ides in aquifer sediments and depletion of dissolved sulfate in ground water resulted in the onset of methanogenesis. Between 1996 and 2000, water-chemistry data indicated that methanogenic metabolism became increasingly prevalent. Molecular analysis of 16S-rDNA extracted from sediments shows the presence of a more diverse methanogenic community inside as opposed to outside the plume core, and is consistent with water-chemistry data indicating a shift toward methanogenesis over time. This rapid evolution of redox processes reflects several factors including the large amounts of contaminants, relatively rapid ground water flow (approximately 0.3 m/day [approximately foot/day]), and low concentrations of microbially reducible Fe(III) oxyhydroxides ( approximately 1 micromol/g) initially present in aquifer sediments. These results illustrate that, under certain hydrologic conditions, redox conditions in petroleum hydrocarbon-contaminated aquifers can change rapidly in time and space, and that the availability of solid-phase Fe(III)-oxyhydroxides affects this rate of change.

Hydrocarbons↗

Cancer mortality in U.S. counties with petroleum industries.

A survey of cancer mortality from 1950 to 1969 was conducted in U.S. counties where the petroleum industry is most heavily concentrated. Male residents of these counties experienced significantly higher rates for cancers of the lung, the nasal cavity and sinuses, and the skin (including malignant melanoma) compared to male residents of counties with similar demographic characteristics. Further study is needed to determine whether these patterns result from exposure to chemical carcinogens, including polycyclic hydrocarbons, involved in the manufacturing of petroleum.

Environmental Exposure↗

Microbial degradation of n-alkyl tetrahydrothiophenes found in petroleum.

Although n-alkyl-substituted tetrahydrothiophenes are found in nonbiodegraded petroleums, they are not found in petroleums which have undergone biodegradation in their reservoirs. These observations suggested that this group of compounds with alkyl chain lengths from approximately C10 to at least C30 is biodegradable. Two of these sulfides, 2-n-dodecyltetrahydrothiophene (DTHT) and 2-n-undecyltetrahydrothiophene, were synthesized, and their biodegradabilities were tested by using five gram-positive, n-alkane-degrading bacterial isolates. The alkyl side chains of these compounds were oxidized, and the major intermediates found in 2-n-undecyltetrahydrothiophene- and DTHT-metabolizing cultures were 2-tetrahydrothiophenecarboxylic acid (THTC) and 2-tetrahydrothiopheneacetic acid (THTA), respectively. Four n-alkane-degrading fungi were also shown to degrade DTHT, yielding both THTA and THTC. Quantitation of tetrahydrothiophene ring-containing products in 28-day-old bacterial and fungal cultures suggested that THTC and THTA were metabolized further to unidentified products. In addition, two of the bacterial isolates were shown to degrade a mixture of n-alkyl tetrahydrothiophenes isolated from Bellshill Lake crude oil.

Biodegradation, Environmental↗

Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities.

Petroleum-contaminated groundwater discharged from underground crude oil storage cavities (cavity groundwater) harbored more than 10(6) microorganisms ml(-1), a density 100 times higher than the densities in groundwater around the cavities (control groundwater). To characterize bacterial populations growing in the cavity groundwater, 46 PCR-amplified almost full-length 16S ribosomal DNA (rDNA) fragments were cloned and sequenced, and 28 different sequences were obtained. All of the sequences were affiliated with the Proteobacteria; 25 sequences (43 clones) were affiliated with the epsilon subclass, 2 were affiliated with the beta subclass, and 1 was affiliated with the delta subclass. Two major clusters (designated clusters 1 and 2) were found for the epsilon subclass proteobacterial clones; cluster 1 (25 clones) was most closely related to Thiomicrospira denitrificans (88% identical in nucleotide sequence), while cluster 2 (11 clones) was closely related to Arcobacter spp. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rDNA fragments showed that one band was detected most strongly in cavity groundwater profiles independent of storage oil type and season. The sequence of this major band was identical to the sequences of most of the cluster 1 clones. Fluorescence in situ hybridization (FISH) indicated that the cluster 1 population accounted for 12 to 24% of the total bacterial population. This phylotype was not detected in the control groundwater by DGGE and FISH analyses. These results indicate that the novel members of the epsilon subclass of the Proteobacteria grow as major populations in the petroleum-contaminated cavity groundwater.

Cloning, Molecular↗

Petroleum pollution bioremediation using water-insoluble uric acid as the nitrogen source.

The biodegradation of hydrocarbon pollutants in open systems is limited by the availability of a utilizable nitrogen source. This limitation can be overcome by using uric acid. Enrichment cultures grown on crude oil-uric acid media yielded mixed and pure cultures that degraded petroleum. In a simulated open system, uric acid bound to crude oil and was available for bacterial growth and petroleum biodegradation.

Acinetobacter↗

Activity and diversity of methanogens in a petroleum hydrocarbon-contaminated aquifer.

Methanogenic activity was investigated in a petroleum hydrocarbon-contaminated aquifer by using a series of four push-pull tests with acetate, formate, H(2) plus CO(2), or methanol to target different groups of methanogenic Archaea. Furthermore, the community composition of methanogens in water and aquifer material was explored by molecular analyses, i.e., fluorescence in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes amplified with the Archaea-specific primer set ARCH915 and UNI-b-rev, and sequencing of DNA from dominant DGGE bands. Molecular analyses were subsequently compared with push-pull test data. Methane was produced in all tests except for a separate test where 2-bromoethanesulfonate, a specific inhibitor of methanogens, was added. Substrate consumption rates were 0.11 mM day(-1) for methanol, 0.38 mM day(-1) for acetate, 0.90 mM day(-1) for H(2), and 1.85 mM day(-1) for formate. Substrate consumption and CH(4) production during all tests suggested that at least three different physiologic types of methanogens were present: H(2) plus CO(2) or formate, acetate, and methanol utilizers. The presence of 15 to 20 bands in DGGE profiles indicated a diverse archaeal population. High H(2) and formate consumption rates agreed with a high diversity of methanogenic Archaea consuming these substrates (16S rRNA gene sequences related to several members of the Methanomicrobiaceae) and the detection of Methanomicrobiaceae by using FISH (1.4% of total DAPI [4',6-diamidino-2-phenylindole]-stained microorganisms in one water sample; probe MG1200). Considerable acetate consumption agreed with the presence of sequences related to the obligate acetate degrader Methanosaeata concilii and the detection of this species by FISH (5 to 22% of total microorganisms; probe Rotcl1). The results suggest that both aceticlastic and CO(2)-type substrate-consuming methanogens are likely involved in the terminal step of hydrocarbon degradation, while methanogenesis from methanol plays a minor role. DGGE profiles further indicate similar archaeal community compositions in water and aquifer material. The combination of hydrogeological and molecular methods employed in this study provide improved information on the community and the potential activity of methanogens in a petroleum hydrocarbon-contaminated aquifer.

Archaea↗

Molecular characterization of resistance-nodulation-division transporters from solvent- and drug-resistant bacteria in petroleum-contaminated soil.

PCR assays for analyzing resistance-nodulation-division transporters from solvent- and drug-resistant bacteria in soil were developed. Sequence analysis of amplicons showed that the PCR successfully retrieved transporter gene fragments from soil. Most of the genes retrieved from petroleum-contaminated soils formed a cluster (cluster PCS) that was distantly related to known transporter genes. Competitive PCR showed that the abundance of PCS genes is increased in petroleum-contaminated soil.

Anti-Bacterial Agents↗

Evaluation of petroleum-degrading potential of bacteria from water and sediment.

Bacteria from water and sediment of an oil-polluted harbor were examined for ability to degrade petroleum. Water samples contained a greater variety of bacterial species capable of degrading petroleum than sediment. Cultures from both water and sediment contained Pseudomonas and Acinetobacter spp. Bacteria present in the water samples produced significantly greater degradation of 2-,3-,4-, and 5-ring cycloalkanes and mono-, di-, tri-, tetra-, and pentaaromatics compared with bacteria in sediment samples.

Acinetobacter↗