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Compound-specific carbon and hydrogen isotope analysis of sub-parts per billion level waterborne petroleum hydrocarbons.

Compound-specific carbon and hydrogen isotope analysis (CSCIA and CSHIA) has been increasingly used to study the source, transport, and bioremediation of organic contaminants such as petroleum hydrocarbons. In natural aquatic systems, dissolved contaminants represent the bioavailable fraction that generally is of the greatest toxicological significance. However, determining the isotopic ratios of waterborne hydrophobic contaminants in natural waters is very challenging because of their extremely low concentrations (often at sub-parts ber billion, or even lower). To acquire sufficient quantities of polycyclic aromatic hydrocarbons with 10 ng/L concentration for CSHIA, more than 1000 L of water must be extracted. Conventional liquid/liquid or solid-phase extraction is not suitable for such large volume extractions. We have developed a new approach that is capable of efficiently sampling sub-parts per billion level waterborne petroleum hydrocarbons for CSIA. We use semipermeable membrane devices (SPMDs) to accumulate hydrophobic contaminants from polluted waters and then recover the compounds in the laboratory for CSIA. In this study, we demonstrate, under a variety of experimental conditions (different concentrations, temperatures, and turbulence levels), that SPMD-associated processes do not induce C and H isotopic fractionations. The applicability of SPMD-CSIA technology to natural systems is further demonstrated by determining the delta13C and deltaD values of petroleum hydrocarbons present in the Pawtuxet River, RI. Our results show that the combined SPMD-CSIA is an effective tool to investigate the source and fate of hydrophobic contaminants in the aquatic environments.

Alkanes↗

Degradation of petroleum model compounds immobilized on clay by a hypersaline microbial mat.

In this study the degradation of hydrophobic petroleum model compounds (phenanthrene, pristane, octadecane and dibenzothiophene) added to a submersed hypersaline microbial mat was investigated. Montmorillonite with an artificially altered, hydrophobic surface was used as carrier material, forming an organo-clay complex (OCC) with the attached mixture of petroleum model compounds. 6 mg/cm2 OCC were applied to cyanobacterial mat pieces, containing approximately 33.3 microg/mg OCC of each compound. The degradation experiment was performed under controlled laboratory conditions and accompanied by chemical analyses by GC/GC-MS, molecular analyses by PCR and DGGE as well as functional analyses by microsensor measurements of oxygen, photosynthesis, sulfide, pH and light. All applied model compounds were degraded, but residues were still present after 18 weeks. The aromatic compounds phenanthrene (5.1 microg/mg OCC) and dibenzothiophene (4.3 microg/mg OCC) were preferentially degraded compared to the alkanes pristane (12.4 microg/mg OCC) and n-octadecane (13.4 microg/mg OCC). Metabolic changes during the degradation process could not be detected by microsensor measurements. The molecular population analyses did not reveal any significant community changes concomitant with the decrease of the petroleum model compounds. We conclude, that the pristine mats represent an intact, robust ecosystem in which the enzymatic requirements for the degradation of the applied pollutants exist. The slow degradation process did not affect the usual high internal turnover rates and did not favor a certain population in the community of the mats.

Aluminum Silicates↗

Organic-inorganic interactions in petroleum-producing sedimentary basins.

Petroleum deposits form as a consequence of the increased temperatures that accompany progressive burial of organic matter deep within sedimentary basins. Recent advances in petroleum geochemistry suggest that inorganic sedimentary components participate in organic transformations associated with this process. Water is particularly important because it facilitates reaction mechanisms not available in dry environments, and may contribute hydrogen and oxygen for the formation of hydrocarbons and oxygenated alteration products. These findings suggest that petroleum generation and stability is influenced by subsurface chemical environments, and is a simple function of time, temperature and the composition of sedimentary organic matter.

Evolution, Chemical↗

In situ laser-induced fluorescence (LIF) analysis of petroleum product-contaminated soil samples.

General considerations of the calibrations of in situ measurements are presented and the concept of using an "average oil" with average analysability for calibration purposes is introduced. The in situ analysis of 30 petroleum product-contaminated soil samples with laser-induced fluorescence (LIF) spectroscopy was performed. Compared to an uncontaminated laboratory reference (LR) soil, 23 soil samples exhibited significantly higher LIF signals, so that these soil samples were classified as contaminated. The repeatability and reproducibility of the in situ LIF analysis were investigated. For the calibration of the LIF data, two LR oils (a fuel oil and a crude oil) were employed. The degree of soil contamination with petroleum products ranged from the limit of detection (LOD) for LIF analysis (ca. 100 ppm), or below, to more than 10,000 ppm. The petroleum product concentrations determined with in situ LIF analysis reveal a reasonable correlation with the results of standard IR analysis after extraction of the contaminated soils.

Calibration↗

A new rapid technique for screening the potential of implanted microorganisms to tolerate and grow on petroleum oily sludges.

Standard experimental protocols for biodegrading petroleum hydrocarbons employ microbiological and/or analytical analysis, and require advanced machines and specialized professionals to carry them out. Furthermore, the obtained results involve a high margin of error. In this paper, a new experimental technique was developed to facilitate the study of microbial capability to degrade petroleum oily sludges. The technique is based on growing microorganisms on 0.22 microm filter membranes laid over oily sludge placed in metallic cups. The microbial growth was assessed using the plate counts technique. Sludge degradation was assessed using Fourier Transform Infrared Spectrometry (FTIR), and compared to the decrease in total petroleum hydrocarbons (TPH) using solvent extraction. The protocol was tested using three types of inocula: fungal inoculum containing Paecilomyces variotii; bacterial inoculum containing Bacillus cereus; and fungal-bacterial inoculum containing both strains. Both, fungal and bacterial, strains were isolated from the oily sludge used in the present study, and were tentatively identified as oil degraders. The results of the newly developed technique helped to assess the potential of these cultures to tolerate and grow on the oily sludge.

Bacillus cereus↗

Petroleum mineral oil refining and evaluation of cancer hazard.

Petroleum base oils (petroleum mineral oils) are manufactured from crude oils by vacuum distillation to produce several distillates and a residual oil that are then further refined. Aromatics including alkylated polycyclic aromatic compounds (PAC) are undesirable constituents of base oils because they are deleterious to product performance and are potentially carcinogenic. In modern base oil refining, aromatics are reduced by solvent extraction, catalytic hydrotreating, or hydrocracking. Chronic exposure to poorly refined base oils has the potential to cause skin cancer. A chronic mouse dermal bioassay has been the standard test for estimating carcinogenic potential of mineral oils. The level of alkylated 3-7-ring PAC in raw streams from the vacuum tower must be greatly reduced to render the base oil noncarcinogenic. The processes that can reduce PAC levels are known, but the operating conditions for the processing units (e.g., temperature, pressure, catalyst type, residence time in the unit, unit engineering design, etc.) needed to achieve adequate PAC reduction are refinery specific. Chronic dermal bioassays provide information about whether conditions applied can make a noncarcinogenic oil, but cannot be used to monitor current production for quality control or for conducting research or developing new processes since this test takes at least 78 weeks to conduct. Three short-term, non-animal assays all involving extraction of oil with dimethylsulfoxide (DMSO) have been validated for predicting potential carcinogenic activity of petroleum base oils: a modified Ames assay of a DMSO extract, a gravimetric assay (IP 346) for wt. percent of oil extracted into DMSO, and a GC-FID assay measuring 3-7-ring PAC content in a DMSO extract of oil, expressed as percent of the oil. Extraction with DMSO concentrates PAC in a manner that mimics the extraction method used in the solvent refining of noncarcinogenic oils. The three assays are described, data demonstrating the validation of the assays are shown, and test results of currently manufactured base oils are summarized to illustrate the general lack of cancer hazard for the base oils now being manufactured.

Animals↗

Total petroleum hydrocarbons and trace metals in street dusts from Tshwane Metropolitan area, South Africa.

This study reports the level of total petroleum hydrocarbons (TPHs) and trace metals in dusts from some petroleum handling facilities (gasoline stations), selected high traffic density roads and residential areas within the Tshwane Metropolitan area in South Africa. Total petroleum hydrocarbons in dust samples were extracted using soxhlet extraction and isolated gravimetrically after column cleanup, while total trace metals in dust samples were digested using mineral acid digestion and analyzed using atomic absorption spectrometry (AAS). The mean TPH level from the residential sites ranged from 206 +/- 20.3-300 +/- 36.4 microg g(-1) while those from gasoline stations and high traffic density roads varied between 562 +/- 43.9-340 +/- 62.6 microg g(-1) and 404 +/- 32.7-852 +/- 55.4 microg g(-1) respectively. Mean concentrations of analyzed trace metals at residential areas ranged from 0.04 +/- 0.02-0.07 +/- 0.03 microg g(-1); ND; 0.21 +/- 0.01-0.34 +/- 0.02 microg g(-1); 0.66 +/- 0.06-2.11 +/- 0.82 microg g(-1); 18.7 +/- 0.86-33.4 +/- 0.83 microg g(-1) and 0.07 +/- 0.04-0.23 +/- 0.01 microg g(-1) for Cd, Pb, Cu, Zn, Mn and Cr respectively. Mean levels from gasoline stations ranged between 0.05 +/- 0.02-0.17 +/- 0.07; 0.08 +/- 0.02-0.12 +/- 0.04; 0.21 +/- 0.04-4.34 +/- 0.60; 7.44 +/- 0.40-13.0 +/- 0.56; 23.36 +/- 4.37-131 +/- 10.5 and 0.05 +/- 0.02-0.17 +/- 0.05 microg g(-1) while those from high traffic density roads ranged from 0.07 +/- 0.04-0.10 +/- 0.04; ND-0.30 +/- 0.05; 0.67 +/- 0.05-3.80 +/- 0.82; 2.40 +/- 0.25-10.6 +/- 0.96; 19.8 +/- 1.03-108 +/- 4.44 and 0.06 +/- 0.02-0.12 +/- 0.04 microg g(-1) also in the above metal order. Contamination by TPH and trace metals both at gasoline stations and high traffic density roads is revealed by their significantly higher values compared to those of residential areas.

Cities↗

Increased risk of preterm delivery in areas with air pollution from a petroleum refinery plant in Taiwan.

The petrochemical and petroleum industries are among the main sources of industrial air pollution in Taiwan. Data in this study concern outdoor air pollution and the health of individuals living in communities in close proximity to a petroleum refinery plant. The prevalence of delivery of preterm birth infants was significantly higher in mothers living in a petroleum refinery area compared to controls in Taiwan. After controlling for several possible confounders (including maternal age, season, marital status, maternal education, and infant sex), the adjusted odds ratio was 1.41 (95% CI = 1.08-1.82) for delivery of preterm infants in the polluted region. Data support the view that air pollution can affect the outcome of pregnancy.

Adult↗

Embryotoxic effects of benzo[a]pyrene, chrysene, and 7,12-dimethylbenz[a]anthracene in petroleum hydrocarbon mixtures in mallard ducks.

Studies with different avian species have revealed that surface applications of microliter amounts of some crude and fuel oils that coat less than 10% of the egg surface result in considerable reduction in hatching with teratogenicity and stunted growth. Other studies have shown that the embryotoxicity is dependent on the aromatic hydrocarbon content, further suggesting that the toxicity is due to causes other than asphyxia. In the present study the effects of three polycyclic aromatic hydrocarbons identified in petroleum were examined on mallard (Anas platyrhynchos) embryo development. Addition of benzo[a]pyrene (BaP), chrysene, or 7,12-dimethylbenz[a]anthracene (DMBA) to a synthetic petroleum hydrocarbon mixture of known composition and relatively low embryotoxicity resulted in embryotoxicity that was enhanced or equal to that of crude oil when 10 microliter was applied externally to eggs at 72 h of development. The order of ability to enhance embryotoxicity was DMBA greater than BaP greater than chrysene. The temporal pattern of embryonic death was similar to that reported after exposure to crude oil, with additional mortality occurring after outgrowth of the chorioallantois. Retarded growth, as reflected by embryonic body weight, crown-rump length, and bill length, was accompanied by teratogenicity. Abnormal embryos exhibited extreme stunting; eye, brain, and bill defects; and incomplete ossification. Gas chromatographic-mass spectral analysis of externally treated eggs showed the passage of aromatic hydrocarbons including chrysene through the shell and shell membranes to the developing embryos. These findings suggest that the presence of polycyclic aromatic hydrocarbons in petroleum, including BaP, chrysene, and DMBA, significantly enhances the overall embryotoxicity in avian species.

9,10-Dimethyl-1,2-benzanthracene↗

Radiological assessment of the level of safety in logging operations in the Nigerian petroleum industry.

Petroleum prospecting and producing activities have been going on in the Niger Delta area of Nigeria for about 40 years. During this period controlled substances such as chemicals and radioactive materials have been widely used in petroleum exploration and exploitation. Deviations from acceptable levels of certain parameters relevant to safety and environmental protection have been encountered, but most have not been investigated or documented. In particular, cases involving the unsafe use, loss and abandonment of radioactive materials have neither received the desired attention nor been reported. This work reports a radiological assessment of safety in the use of radioactive and radiation producing materials in logging and well study operations in the Nigerian petroleum industry. The assessment protocol used for the evaluation is based on a numerical ranking system. Based on a scale of 100, it is found in this logging and well study that the level of safety as defined in the text is around 60% for all three sites assessed. There is substantial work needed to raise the radiation protection standards further at these sites.

Humans↗

Work-related musculoskeletal disorders in Norway's offshore petroleum industry.

BACKGROUND: Since 1992, physicians have reported work-related diseases among workers in Norway's offshore petroleum industry to the Petroleum Safety Authority, as required by law. AIMS: To analyse the number of reported work-related musculoskeletal disorders and risk factors (occupation and reported exposure) from 1992 to 2003. METHODS: Data from the Petroleum Safety Authority's registry of work-related diseases were analysed. RESULTS: During the 12 years, 3131 new work-related musculoskeletal disorders were reported and this was the category of work-related disease most frequently reported (47%). The number of work-related musculoskeletal disorders varied substantially from year to year. Disorders of the upper limb accounted for 53% and back disorders for 20% of all work-related musculoskeletal disorders. Lower limb disorders accounted for 16%, of which knee disorders dominated (12% of all cases). The dominant occupational categories were maintenance work (40%) and catering (21%). Frequently reported types of exposure were high physical workload, repetitive work and walking on hard surfaces/climbing stairs and ladders. CONCLUSION: Strategies for preventing musculoskeletal disorders should be carried out to reduce the burden of high physical workload and repetitive work, especially in maintenance work and catering. Further research is recommended on the association between walking on hard surfaces/climbing stairs and ladders and knee disorders. Reporting routines need to be improved to monitor trends over time and to assess the effects of interventions.

Humans↗

Radiographic operations and safety in the Nigerian petroleum industry.

This work reports on a radiation safety study of the use of radioactive sources and radiation-producing machines for radiographic purposes in the Nigerian Petroleum Industry. It is a part of a wider study on the level of safety in the use of radiation in the country-a study that covered the manufacturing and petroleum industries. The survey assessment is based on minimum requirements for radiological safety, on a scale of 0-100, to protect personnel, the public, and the environment from artificial radiation sources. The survey result shows that the level of safety in the use of radiation-emitting machines for radiographic purposes is scored at 52, while it is 66 in the use of radioisotopes. Consequently, the level of safety in radiographic practices in the Nigerian Petroleum Industry is about 60% of the ideal.

Extraction and Processing Industry↗

Bacterial diversity of a cyanobacterial mat degrading petroleum compounds at elevated salinities and temperatures.

Cyanobacterial mats of the Arabian Gulf coast of Saudi Arabia experience extreme conditions of temperature and salinity. Because they are exposed to continuous oil pollution, they form ideal models for biodegradation under extreme conditions. We investigated the bacterial diversity of these mats using denaturing gradient gel electrophoresis and 16S rRNA cloning, and tested their potential to degrade petroleum compounds at various salinities (fresh water to 16%) and temperatures (5 to 50 degrees C). Cloning revealed that c. 15% of the obtained sequences were related to unknown, possibly novel bacteria. Bacteria belonging to Beta-, Gamma- and Deltaproteobacteria, Cytophaga-Flavobacterium-Bacteroides group and Spirochetes, were detected. The biodegradation of petroleum compounds at different salinities by mat microorganisms showed that pristine and n-octadecane were optimally degraded at salinities between 5 and 12% (weight per volume NaCl) whereas the optimum degradation of phenanthrene and dibenzothiophene was at 3.5% salinity. The latter compounds were also degradable at 8% salinity. The same compounds were degraded at temperatures between 15 and 40 degrees C but not at 5 and 50 degrees C. The optimum temperature of degradation was 28-40 degrees C for both aliphatics and aromatics. We conclude that the studied microbial mats from Saudi Arabia are rich in novel halotolerant and thermotolerant microorganisms with the potential to degrade petroleum compounds at elevated salinities and temperatures.

Anti-Bacterial Agents↗

Molecular analysis of the bacterial community in a continental high-temperature and water-flooded petroleum reservoir.

Water from a continental high-temperature, long-term water-flooded petroleum reservoir in Huabei Oilfield in China was analysed for its bacterial community and diversity. The bacteria were characterized by their 16S rRNA genes. A 16S rRNA gene clone library was constructed from the community DNA, and using restriction fragment length polymorphism analysis, 337 randomly selected clones were clustered with 74 operational taxonomic units. Sequencing and phylogenetic analyses showed that the screened clones were affiliated with Gammaproteobacteria (85.7%), Thermotogales (6.8%), Epsilonproteobacteria (2.4%), low-G+C Gram-positive (2.1%), high-G+C Gram-positive, Betaproteobacteria and Nitrospira (each <1.0%). Thermopilic bacteria were found in the high-temperature water from the flooded petroleum reservoir, as well as mesophilic bacteria such as Pseudomonas-like clones. The mesophilic bacteria were probably introduced into the reservoir as it was being exploited. This work provides significant information on the structure of bacterial communities in high-temperature, long-term water-flooded petroleum reservoirs.

Bacteria↗

Sequential electron acceptor model for evaluation of in situ bioremediation of petroleum hydrocarbon contaminants in groundwater.

Mathematical development and model application is provided for a multiple substrate, sequential electron acceptor model, accounting for hydrodynamic transport, adsorption, and sequential oxygen/iron(III)-based biodegradation. Equations for iron(III)-based biodegradation of petroleum hydrocarbons are developed based on oxygen-inhibited Monod kinetics. The iron(III)-based biodegradation expressions were combined with earlier work by Widdowson and Aelion, to develop the two-dimensional, multiple substrate, oxygen/iron(III) sequential electron acceptor biodegradation model presented here. In addition to mathematical model development, simulations demonstrating the advantages of sequential electron acceptor and multiple substrate biodegradation models are provided. These simulations show that commonly-used single electron acceptor models may underpredict natural, in situ biodegradation potential at sites where indigenous microorganisms are capable of using multiple electron acceptors. Additional simulations show that, for contaminant plumes composed of constituents which biodegrade at different rates and under varying electron acceptor conditions, a multiple substrate model may allow more accurate prediction of both individual contaminant concentrations and the total amount of biodegraded contaminant. Considering that typical contaminant plumes are composed of multiple constituents with varying biodegradation properties and health risks, multiple substrate sequential electron acceptor models have the potential to provide more accurate tracking of individual constituent migration. The model was applied to a leaking UST site in Laurel Bay, South Carolina. Laboratory and monitoring well data presented in Landmeyer et al. have established that the petroleum hydrocarbon contaminants are present in the groundwater and are undergoing sequential oxygen-iron(III)-based biodegradation. Model simulations proved capable of reproducing the trends observed at the Laurel Bay site in that BTX contaminants were removed by sequential biodegradation, occurring first aerobically and subsequently anaerobically, and that iron(III)-reducing organisms biodegrade contaminants only in the absence of oxygen. The BTX compounds were individually but simultaneously modeled, allowing explicit modeling of specific contaminant biodegradation properties (e.g., toluene and xylene are assumed to degrade sequentially and benzene is assumed to degrade aerobically only). Although simulations presented here can reproduce trends observed at the Laurel Bay site, inclusion of additional electron acceptors and additional model calibration to data from this and other sites is necessary to improve and verify the model's capability to predict the efficacy of intrinsic biodegradation of petroleum hydrocarbon contaminants in groundwater.

Bacteria↗

Numerical taxonomy and ecology of petroleum-degrading bacteria.

A total of 99 strains of petroleum-degrading bacteria isolated from Chesapeake Bay water and sediment were identified by using numerical taxonomy procedures. The isolates, together with 33 reference cultures, were examined for 48 biochemical, cultural, morphological, and physiological characters. The data were analyzed by computer, using both the simple matching and the Jaccard coefficients. Clustering was achieved by the unweighted average linkage method. From the sorted similarity matrix and dendrogram, 14 phenetic groups, comprising 85 of the petroleum-degrading bacteria, were defined at the 80 to 85% similarity level. These groups were identified as actinomycetes (mycelial forms, four clusters), coryneforms, Enterobacteriaceae, Klebsiella aerogenes, Micrococcus spp. (two clusters), Nocardia species (two clusters), Pseudomonas spp. (two clusters), and Sphaerotilus natans. It is concluded that the degradation of petroleum is accomplished by a diverse range of bacterial taxa, some of which were isolated only at given sampling stations and, more specifically, from sediment collected at a given station.

Bacteria↗

Selective removal of nitrogen from quinoline and petroleum by Pseudomonas ayucida IGTN9m.

Enrichment culture experiments employing soil and water samples obtained from petroleum-contaminated environments succeeded in the isolation of a pure culture possessing the ability to utilize quinoline as a sole nitrogen source but did not utilize quinoline as a carbon source. This culture was identified as Pseudomonas ayucida based on a partial 16S rRNA gene sequence, and the strain was given the designation IGTN9m. Examination of metabolites using thin-layer chromatography and gas chromatography-mass spectrometry suggests that P. ayucida IGTN9m converts quinoline to 2-quinolinone and subsequently to 8-hydroxycoumarin. Resting cells of P. ayucida IGTN9m were shown to be capable of selectively removing about 68% of quinoline from shale oil in a 16-h treatment time. These results suggest that P. ayucida IGTN9m may be useful in petroleum biorefining for the selective removal of organically bound nitrogen from petroleum.

Biodegradation, Environmental↗

Microbial diversity of a heavily polluted microbial mat and its community changes following degradation of petroleum compounds.

We studied the microbial diversity of benthic cyanobacterial mats inhabiting a heavily polluted site in a coastal stream (Wadi Gaza) and monitored the microbial community response induced by exposure to and degradation of four model petroleum compounds in the laboratory. Phormidium- and Oscillatoria-like cyanobacterial morphotypes were dominant in the field. Bacteria belonging to different groups, mainly the Cytophaga-Flavobacterium-Bacteriodes group, the gamma and beta subclasses of the class Proteobacteria, and the green nonsulfur bacteria, were also detected. In slurry experiments, these communities efficiently degraded phenanthrene and dibenzothiophene completely in 7 days both in the light and in the dark. n-Octadecane and pristane were degraded to 25 and 34% of their original levels, respectively, within 7 days, but there was no further degradation until 40 days. Both cyanobacterial and bacterial communities exhibited noticeable changes concomitant with degradation of the compounds. The populations enriched by exposure to petroleum compounds included a cyanobacterium affiliated phylogenetically with Halomicronema. Bacteria enriched both in the light and in the dark, but not bacteria enriched in any of the controls, belonged to the newly described Holophaga-Geothrix-Acidobacterium phylum. In addition, another bacterial population, found to be a member of green nonsulfur bacteria, was detected only in the bacteria treated in the light. All or some of the populations may play a significant role in metabolizing the petroleum compounds. We concluded that the microbial mats from Wadi Gaza are rich in microorganisms with high biodegradative potential.

Alkanes↗