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Feasibility study of bioremediation of a drilling-waste-polluted soil: stimulation of microbial activities and hydrocarbon removal.

The objective of this study was to determine the feasibility of bioremediation as a treatment option for an aged and chronically polluted drilling waste soil located at the Southeast of Mexico. The polluted drilling-waste site with a mean total petroleum hydrocarbon concentration (TPHs) of 39,397 +/- 858 mg/kg was treated with one dose of a nutrient-surfactant commercial product at 40 mg/kg soil and two doses of H2O2 (50 and 100 mg H2O2/kg soil). In this study, the parameters that were monitored include soil respiration, heterotrophic and hydrocarbon-degrading bacteria as biological indicators, catalase and dehydrogenase activities, and TPHs degradation as decontamination parameters. The results demonstrated that the microbial activities can be stimulated in a polluted drilling-waste site by the addition of H2O2 and commercial product, thereby resulting in increasing TPHs degradation. These aspects must be taken into account when biodegradation studies involve the application of a commercial product.

Analysis of Variance↗

Determination of annatto in high-fat dairy products, margarine and hard candy by solvent extraction followed by high-performance liquid chromatography.

Utilizing solvents such as ethanolic aqueous ammonia, petroleum ether, hexane and chloroform, annatto components alpha- and beta-norbixin and alpha- and beta-bixin were extracted from cheese, butter, margarine and hard candy. After transferring the extract into a solution of aqueous acetic acid in methanol, bixin and norbixin were determined quantitatively using high-performance liquid chromatography (HPLC) and an absorbance detector set at 500 nm. Recovery of norbixin from spiked cheese samples averaged 92.6% over a range of 1 to 110 micrograms/g. Commercial cheese samples were found to contain 1.1-68.8 microgram/g total norbixin, and two samples also contained 5.1-5.6 micrograms/g total bixin. Samples of uncoloured butter were spiked with bixin and recovery averaged 93.2% over a range of 0.1 to 445 micrograms/g. Levels of 0.2 microgram/g total bixin and 0.91 microgram/g total norbixin were found in one commercial butter sample; the others contained trace levels of both compounds. Hard candies were prepared in the laboratory and recovery studies conducted. Recovery of norbixin averaged 88%.

Bixaceae↗

Bacterial sulfate production by biodesulfurization of aromatic hydrocarbons, determined by ion chromatography.

The use of bacteria to remove sulfur from crude oil or petroleum distillates is a novel concept that presents an alternative biotechnology to the current technology of hydrodesulfurization (HDS). Sulfur must be removed from crude oils prior use. The burning of fossil fuels containing sulfur releases sulfur dioxide into the atmosphere causing acid rain. The aim of this work is to determine the sulfate concentration by ion chromatography (IC), and calculate the percentage of transformation of organic bound sulfur, that is converted to sulfate, and estimate the efficiency of bacteria in desulfurization. IC is a suitable method for sulfate concentration determination. However, when chloride concentrations are significantly high, interference of the sulfate signal does occur. In this case, it could be avoided by diluting samples. A Dionex Model 2000i/SP IC system, with an anionic pre-column (Dionex AG4A), an anion separator column (Dionex AS4A), a suppressor column (Dionex AMMS-II), and a conductivity detector was used. The eluent (21 mM NaOH) and regenerant (electrolyzed 18 M omega/cm water) flow-rates were 1.0 and 2.0 ml/min, respectively. The sample loop volume was 10 microliters and the conductivity sensitivity was 30 muS. The diluted samples were filtered through a 0.45-micron filter before injection. The highest sulfate concentration detected was 24.10 mg/l, corresponding to a maximal conversion rate of 10% in a month. Sulfate ions were not detected in control samples. The correlation coefficient for a linear least squares fit was 0.99 (p < 0.001). The minimal concentration that we can read was 0.02 mg/l and this concentration corresponded to the limit of detection obtained under the conditions employed in this study. IC is an economical, sensitive and accurate way to estimate the sulfate concentrations in microbiological samples.

Bacillus↗

[Development and testing of "Ekosorb" biosorbent based on association of petroleum-oxidizing bacteria for purifying petroleum-polluted soils].

A biosorbent containing an association of oil-oxidizing bacteria as a main constituent was developed, in which Lessorb, a product of moss and wood thermal processing, was used as a carrier. Xeroprotectors preserving the cell viability and oil-oxidizing activity in the biosorbent on drying and after long-term storage were selected. The use of this biosorbent for cleaning oil-polluted sod-podzol soils showed a two-threefold cleanup rate acceleration at different pollution levels (8 and 24 l/m2), especially in the presence of a nitrogen-phosphate fertilizer. The biosorbent increased the populations of certain groups of soil microorganisms and the total soil biological activity.

Absorption↗

Biodesulfurization and the upgrading of petroleum distillates.

Biotechnology offers an alternative way to process fossil fuels. There have been several important advances in the elucidation of the mechanisms of biodesulfurization and the development of a biocatalytic desulfurization process. These include a detailed analysis of the rate and extent of desulfurization of real target molecules in a diesel matrix, the directed evolution of rate- and extent-limiting enzymes for better performance and the expression of the genes in alternative hosts. Process innovations include new reactor designs, separations and recovery strategies and the production of value-added byproducts during desulfurization.

Bacteria↗

Microbial desulfurization of organic sulfur compounds in petroleum.

Sulfur removal from petroleum is important from the standpoint of the global environment because the combustion of sulfur compounds leads to the production of sulfur oxides, which are the source of acid rain. As the regulations for sulfur in fuels become more stringent, the existing chemical desulfurizations are coming inadequate for the "deeper desulfurization" to produce lower-sulfur fuels without new and innovative processes. Biodesulfurization is rising as one of the candidates. Several microorganisms were found to desulfurize dibenzothiophene (DBT), a representative of the organic sulfur compounds in petroleum, forming a sulfur-free compound, 2-hydroxybiphenyl. They are promising as biocatalysts in the microbial desulfurization of petroleum because without assimilation of the carbon content, they remove only sulfur from the heterocyclic compounds which is refractory to conventional chemical desulfurization. Both enzymological and molecular genetic studies are now in progress for the purpose of obtaining improved desulfurization activity of organisms. The genes involved in the sulfur-specific DBT desulfurization were identified and the corresponding enzymes have been investigated. From the practical point of view, it has been proved that the microbial desulfurization proceeds in the presence of high concentrations of hydrocarbons, and more complicated DBT analogs are also desulfurized by the microorganisms. This review outlines the progress in the studies of the microbial desulfurization from the basic and practical point of view.

Biodegradation, Environmental↗

Determination of diclofop-methyl and diclofop residues in soil and crops by gas chromatography.

A method is described for the determination of residues of the herbicide diclofop-methyl and its metabolite, diclofop, in soil and crops. The residues were extracted with acetone-light petroleum and extracts were concentrated (diclofop was derivatized to its pentafluorobenzyl derivative), and then the products were purified on a chromatographic column containing alumina, silver-alumina and Florisil. Finally, they were determined by gas chromatography using an electron-capture detector. The detection limits of diclofop-methyl and diclofop were between 0.01 and 0.05 mg/kg. The average recoveries were 76.4-97.2% and 72.8-105.2%, respectively, making the method suitable for statutory residue testing purposes.

Agriculture↗

Disposal options and case-study pathway analyses.

This paper presents an overview of alternatives that currently are employed for the disposal of wastes containing enhanced concentrations of naturally occurring radioactive materials (NORM). The application of pathway assessment methodology to the derivation of acceptable cleanup criteria for residual NORM is described. A specific example is provided drawing on experience with a site containing above-background levels of natural uranium-series and thorium-series nuclides. The options for safe, permanent disposal of U and Th bearing NORM waste are discussed and the application of pathway analysis to performance assessment of a typical engineered disposal cell is illustrated. Typical methods for disposal of other NORM waste forms also are described. This includes radium-contaminated wastes from petroleum sludges, uranium milling, and geothermal power plant operation as well as thorium wastes from production of thorium-magnesium alloys.

Background Radiation↗

Environmental biotechnology: biotechnology solutions for a global environmental problem, hazardous chemical wastes.

Biotechnology has a growing place in the remediation of hazardous waste sites throughout the world, and especially in Asia where population density is high and land and fresh water are scarce. In-situ bioremediation has been demonstrated already to be highly effective for petroleum hydrocarbons (alkanes, aromatics, polychlorophenols) and organophosphate pesticides in soils and for gasoline by-products (benzene, toluene, xylene) and chlorinated solvents (trichloroethylene) in groundwater. Heavy metals and PCBs are not suitable for bioremediation. Environmental biotechnology includes solid-phase and slurry-phase bioremediation for contaminated soils and site-specific bioreactors for contaminated groundwater. Specific examples are presented. From a policy point of view, accumulated wastes must be detoxified, preferably at sites where they already exist. We cannot continue to rely on their removal and disposal "elsewhere". For current waste streams, we must minimize the volumes and toxicity. Environmental biotechnology will play a key role.

Biotechnology↗

Validation of exposure estimation for benzene in the Australian petroleum industry.

A nested case-control study was undertaken to investigate whether an excess of lympho-haematopoietic cancers in the Australian petroleum industry was associated with benzene exposure. The benzene exposures of the cases and controls were estimated using a quantitative algorithm based largely on exposures measured in the Australian petroleum industry. The algorithm was used to estimate, for each subject, the benzene exposure in parts per million (ppm) for each job held in the industry, and the cumulative exposure in ppm years. Because of the critical importance of the exposure assessment in this design of epidemiological study, particular attention was paid to the reliability of the inputs to the algorithm. The inputs [base estimates (BEs) of exposure and technology-specific exposure modifiers (EMs)] were compared to data from other sources including the occupational hygiene literature. Where such comparison data were available, they were generally found to confirm the values used in the algorithm, although four input values were changed as a result of the validation exercise. The integrity of the task-based algorithm was validated by employing it to calculate the exposures of the tanker drivers in the study and comparing these with measured daily exposure for tanker drivers in the Australian petroleum industry and exposure values found in the occupational hygiene literature. After adjustment for the mix of products carried by the Australian tanker drivers, the estimates from the algorithm were found to be comparable to the measured and literature values. This exercise provided evidence that the exposure assessment for the epidemiological study was reliable and that the results of the study can be used as the basis for evaluating the relationship between exposure to benzene and the risk of lympho-haematopoietic cancer.

Algorithms↗

Mist generation from metalworking fluids formulated using vegetable oils.

Metalworking fluid emulsion formulations produced from vegetable oils may be less toxic and may reduce disposal costs when compared with fluids formulated with petroleum-based oils. Experiments were performed on experimental emulsions made with unmodified and modified soybean oils to measure rates of mist production by impaction, centrifugal force and evaporation/recondensation mechanisms. Results were compared with measurements made using a commercial metalworking fluid emulsion formulated using vegetable oil and another made from mineral oil. The results indicated that most of the experimental fluids produced about the same amount of mist as the commercial fluids by impaction and more mist than the petroleum-based fluid by centrifugal force. However, an air-oxidized modified soy oil produced less mist by impaction than the petroleum-based fluid and about the same by centrifugal force. The experimental fluids produced between 30 and 90% less mist than the commercial fluids by evaporation/recondensation. The air-oxidized soybean oil was the most promising candidate among the experimental fluids for further testing in more realistic machining conditions.

Air Pollutants, Occupational↗

Bioconversion of recalcitrant 4-methyldibenzothiophene to water-extractable products using lignin-degrading basidiomycete coriolus versicolor

Under secondary metabolic conditions, the white-rot basidiomycete Coriolus versicolormetabolized 4-methyldibenzothiophene (MDBT), which is a recalcitrant organic sulfur contaminant found in petroleum. The pathway of the transformation of MDBT was elucidated by the identification of fungal metabolites upon the addition of MDBT and its metabolic intermediates. S-oxidation to form MDBT-5-oxide was the initial step of MDBT metabolism. Then, the metabolic pathway was branched to form MDBT-5-dioxide, which was a dead-end product, and hydroxymethylDBT (HMDBT)-5-oxide. Extracellular ligninolytic enzymes such as lignin and manganese peroxidases and laccase did not catalyze the oxidation of either MDBT or MDBT-5-oxide. HMDBT-5-oxide was then oxidized to HMDBT-5-dioxide. Piperonyl butoxide, an inhibitor of cytochrome P450, suppressed fungal oxidation of MDBT to its oxide, MDBT-5-oxide to dioxide and to HMDBT-5-oxide, and HMDBT-5-oxide to dioxide. The efficiency of the inhibition varied for each substrate, suggesting that each oxidation was catalyzed by different enzymes. The hydroxylation of methyl substituents to the hydroxymethyl group was suggested to be catalyzed by a novel monooxygenase. HMDBT-5-dioxide was finally xylosylated most likely by xylosyltrasferase to yield 10-(beta-D-xylopyranosyloxy)-4-methyldibenzothiophene-5-dioxide. The final xyloside product and metabolic intermediates are water-extractable compounds, which would give us a novel strategy for biodesulfurization technology.

Journal Article↗

Utilization of petroleum hydrocarbons by Pseudomonas fluorescens isolated from a petroleum-contaminated soil.

A strain of Pseudomonas fluorescens, isolated from petroleum hydrocarbon-contaminated soil was examined for its ability to utilize a variety of hydrocarbon substrates. Surface tension measurements indicated the production of biosurfactant during the microbial degradation of hydrocarbon. The organism utilized both short and long chain n-alkanes. It emulsified a number of aliphatic and aromatic hydrocarbons.

Biodegradation, Environmental↗

Occupation and the risk of bladder cancer.

The relationship between bladder cancer, occupation and exposure to a number of occupational agents was assessed in a case-control study conducted in the greater Milan area, Northern Italy. The case series consisted of 263 cases (219 males, 44 females) with histologically confirmed invasive bladder cancer, admitted to a network including major teaching and general hospitals in the area under surveillance. The controls were 287 patients (210 males, 77 females), admitted for acute, non-neoplastic or urinary tract diseases to the same network of hospitals. Cases more frequently reported occupation in dyestuff production (relative risk (RR) = 4.6), painting/spraying work (RR = 1.8), chemical industry (RR = 1.7), pharmaceuticals (RR = 1.7) and coal/gas production (RR = 3.1). Only for dyestuff production however, was the excess statistically significant. There was no association with agriculture or related activities, rubber manufacturing, printing, the petroleum industry, food processing and mechanics. In relation to exposure to occupational agents, significant positive trends in risk were observed for dyes/paints (RR = 4.8 for greater than ten years of exposure), herbicides (RR = 4.1), chemicals (RR = 2.4) and gases/fumes (RR = 4.8). No association was found with metals or metal dusts, plastic resins or glues, oil, wood dust, solvents or benzene, asbestos, electricity or radar and coal tar. Besides confirming the well known association between bladder cancer risk and dyestuff production and, to a lesser extent, a wide spectrum of chemical-related activities, this study provides statistically significant evidence of an independent role of herbicides on the risk of bladder cancer.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Mutagenic/carcinogenic agents in indoor pollutants; the dinitropyrenes generated by kerosene heaters and fuel gas and liquefied petroleum gas burners.

Incomplete combustion of kerosene heater, and fuel gas and liquefied petroleum gas-burner emissions produces indoor pollutants that may be carcinogenic. The incomplete-combustion products from each type of appliance were therefore collected by adsorption on about 3 g of XAD-2 resin, and were extracted with benzene-methanol as a solvent for determination and identification of mutagens in the Salmonella-microsome test system. Benzene-methanol extracts of the particulates generated by a heater and two burners showed extreme mutagenicity for strains TA97 and TA98 without S9 mix. Based on the results of analysis, a combination of high performance liquid chromatography (h.p.l.c.) and gas chromatography (GC), about 40-80% of the direct-acting mutagenicity in each crude extract showed the same h.p.l.c. and GC retention times as dinitropyrenes (1,3-, 1,6- and 1,8-isomers), and 1-nitropyrene. Moreover, other nitroarenes, 2-nitrofluorene, 1,5- and 1,8-dinitronaphthalene, and 4,4'-dinitrobiphenyl, were detectable in almost all samples, but their contribution to the mutagenicity of each extract was very low. Kerosene heaters were found to generate small amounts (0.2 ng/h) of dinitropyrenes, which are potential mutagens/carcinogens, only after 1 h of operation.

Air Pollutants↗

Carbon isotopic fractionation associated with lipid biosynthesis by a cyanobacterium: relevance for interpretation of biomarker records.

For the cyanobacterium Synechocystis UTEX 2470, grown photoautotrophically to a logarithmic stage of growth, the total lipid extract is depleted in 13C by 4.8% relative to average biomass. Depletions observed for acetogenic (straight-chain) lipids range from 7.6 (hexadecanoic acid) to 9.9% (a C16 n-alkyl chain bound in a polar-lipid fraction), with a mass-weighted average of 9.1%. Polyisoprenoid lipids fall into two isotopic groups, with phytol, diplopterol, and diploptene depleted by 6.4-6.9% and bishomohopanol (produced from the extracts by the preparative degradation of bacteriohopanepolyol) depleted by 8.4%. Analysis of the pattern of depletions indicates that two carbon positions in each C5 biomonomer leading to polyisoprenoid products are probably depleted in 13C relative to average biomass. The depletion of bacteriohopanepolyol relative to other polyisoprenoids can be ascribed to changes that occur over the life of each cell: (1) the 13C content of carbon flowing to lipid biosynthesis decreases as the cell size increases and (2) a greater proportion of the bacteriohopanepolyol which, unlike other polyisoprenoids, is present mainly in the cytoplasm rather than in membranes and is synthesized when cells are larger. Chlorophyll a is depleted relative to average biomass by O.7%. Given the observed depletion of 13C in phytol, the heteroaromatic, chlorophyllide portion of chlorophyll must be enriched in 13C by 2.7%. This enrichment is large relative to that in chlorophyllides produced by eukaryotes and may be related to a parallel enrichment of 13C in cyanobacterial glutamic acid. As in many previous investigations of cyanobacterial lipids, long-chain n-alkanes (C22-C29) are found in the extracts. They are, however, enriched in 13C relative to biomass and have isotopic compositions suggesting that they are contaminants of petrochemical origin. Available results indicate that cyanobacterial lipids will be depleted relative to dissolved CO2 that has served as a carbon source by 22-30% and that a wider range of depletions will be characteristic of eukaryotic products. The absence of long-chain n-alkanes in cyanobacteria reduces the possibility that petroleum ever formed from pre-eukaryotic sedimentary debris.

Acetic Acid↗