Desulfurization of model and diesel oils by resting cells of Gordona sp.

The desulfurization activity of the resting cells of Gordona sp. CYKS1 was strongly depended on harvest time and the highest value when the cells had been harvested in the early growth phase (0.12 mg sulfur g^–1 cell^–1 h^–1). For the model oil, hexadecane containing dibenzothiophene, the specific desulfurization rate decreased as the reaction proceeded. Both the specific and the volumetric desulfurization rates were not significantly affected by the aqueous-to-oil phase ratio. The diesel oils, light gas oil and a middle distillate unit feed were desulfurized at higher rates (ca. 0.34 mg sulfur g^–1 cell^–1 h^–1) than the model oil (0.12 mg sulfur g^–1 cell^–1 h^–1).     

The use of biomarkers in the antioxidant responses of Daphnia magna to the acute and chronic exposure to no. 20 diesel oil and 2,4-dichlorophenol

Abstract

The effects of no. 20 diesel oil exposure, 2,4-dichlorophenol (2,4-DCP) exposure and combined exposure on the antioxidant defences of Daphnia magna have been studied systematically for the first time. Daphnia magna was exposed for 1 day or 10 days to several concentrations of 0, 0.005, 0.01, 0.05, 0.1, 0.5 or 1.0 mg L^?1 solutions. Antioxidant defences consisting of the levels of reduced glutathione (GSH) and glutathione disulfide (GSSG) and activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), selenium dependent glutathione peroxidase (Se-GPx) and glutathione S-transferase (GST) of daphnids were determined to evaluate their protective roles and to analyse the occurrence of oxidative stress. The possible antioxidant defence mechanisms are discussed. Furthermore, GST can be a potential biomarker and an early-warning index for the pollutants in waters in that GST responded sensitively to 1 day and 10 days of exposure to diesel oil and 2,4-DCP and 10 days of combined exposure. Crossover comparisons showed an antagonistic action about the no-observed-effect concentration (NOEC) against Daphnia magna, which needs further studies.     

Hydrocarbon Degradation and Biosurfactant Production by an Acenaphthene-degrading Pseudomonas Species

An acenaphthene-degrading bacterium putatively identified as Pseudomonas sp. strain KR3 and isolated from diesel-contaminated soil in Lagos, Nigeria was investigated for its degradative and biosurfactant production potentials on crude oil. Physicochemical analysis of the sampling site indicates gross pollution of the soil with high hydrocarbon content (2100 mg/kg) and detection of various heavy metals. The isolate grew luxuriantly on crude oil, engine oil and acenaphthene. It was resistant to septrin, amoxicillin and augmentin but was susceptible to pefloxacin, streptomycin and gentamycin. It was also resistant to elevated concentration of heavy metals such as 1–15 mM lead, nickel and molybdenum. On acenaphthene, the isolate exhibited specific growth rate and doubling time of 0.098 day^?1 and 3.06 days, respectively. It degraded 62.44% (31.2 mg/l) and 91.78% (45.89 mg/l) of 50 mg/l acenaphthene within 12 and 21 days. On crude oil, the specific growth rate and doubling time were 0.375 day^?1 and 1.85 days with corresponding percentage degradation of 33.01% (903.99 mg/l) and 87.79% (2403.71 mg/l) of crude oil (2738.16 mg/l) within 9 and 18 days. Gas chromatographic analysis of residual crude oil at the end of 18 days incubation showed significant reductions in the aliphatic, alicyclic and aromatic fractions with complete disappearance of benzene, propylbenzene, pristane, phytane, and nC₁₈-octadecane fractions of the crude oil. The isolate produced growth-associated biosurfactant on crude oil with the highest emulsification index (E₂₄) value of 72% ± 0.23 on Day 10 of incubation. The partially purified biosurfactant showed zero tolerance for salinity and had its optimal activity at 27°C and pH 2.0.     

A novel qPCR protocol for the specific detection and quantification of the fuel-deteriorating fungus Hormoconis resinae

A wide variety of fungi and bacteria are known to contaminate fuels and fuel systems. These microbial contaminants have been linked to fuel system fouling and corrosion. The fungus Hormoconis resinae, a common jet fuel contaminant, is used in this study as a model for developing innovative risk assessment methods. A novel qPCR protocol to detect and quantify H. resinae in, and together with, total fungal contamination of fuel systems is reported. Two primer sets, targeting the markers RPB2 and ITS, were selected for their remarkable specificity and sensitivity. These primers were successfully applied on fungal cultures and diesel samples demonstrating the validity and reliability of the established qPCR protocol. This novel tool allows clarification of the current role of H. resinae in fuel contamination cases, as well as providing a technique to detect fungal outbreaks in fuel systems. This tool can be expanded to other well-known fuel-deteriorating microorganisms.     

Prepared Bed Land Treatment of Soils Containing Diesel and Crude Oil Hydrocarbons

An ex situ, field-scale, prepared bed land treatment unit (LTU) was used to bio-remediate soils containing petroleum hydrocarbons. Two soils were treated in side-by-side units to compare performance: (1) a clayey silt containing crude oil hydrocarbons from releases 30 to 40 years ago and (2) a silty sand containing diesel fuel hydrocarbons from a leak about three years prior to the bioremediation. The effectiveness of the bioremediation in the LTU was evaluated over a period of 18 months. The results indicated that: (1) prepared bed bioremediation reduced the hydrocarbon concentration, mobility, and relative toxicity in the soil with the diesel fuel, and (2) chemical bioavailability appeared to limit bioremediation of the soil containing the crude oil hydrocarbons. Although the soils containing the crude oil hydrocarbons contained an average of 10,000?mg TPH/kg dry soil, these soils had limited hydrocarbon availability, nontoxic conditions, and low potential for chemical migration. For the soils containing the diesel fuel, active prepared bed bioremediation of about 15 weeks was adequate to reach an environmentally acceptable endpoint. At that time, there was little further TPH loss, no Microtox^TM toxicity, and limited hydrocarbon mobility.     

Repeated inhalations of diesel exhaust particles and oxidatively damaged DNA in young oxoguanine DNA glycosylase (OGG1) deficient mice

DNA repair may prevent increased levels of oxidatively damaged DNA from prolonged oxidative stress induced by, e.g. exposure to diesel exhaust particles (DEP). We studied oxidative damage to DNA in broncho-alveolar lavage cells, lungs, and liver after 4 × 1.5 h inhalations of DEP (20 mg/m³) in Ogg1^- / -  and wild type (WT) mice with similar extent of inflammation. DEP exposure increased lung levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in Ogg1^- / -  mice, whereas no effect on 8-oxodG or oxidized purines in terms of formamidopyrimidine DNA glycosylase (FPG) sites was observed in WT mice. In both unexposed and exposed Ogg1^- / -  mice the level of FPG sites in the lungs was 3-fold higher than in WT mice. The high basal level of FPG sites in Ogg1^- / -  mice probably saturated the assay and prevented detection of DEP-generated damage. In conclusion, Ogg1^- / -  mice have elevated pulmonary levels of FPG sites and accumulate genomic 8-oxodG after repeated inhalations of DEP.     

Evaluating the effects of gross nitrogen mineralization,immobilization, and nitrification on nitrogen fertilizer availability in soil experimentally contaminated with diesel

Sandy clay loam soil was contaminated with 5000 mg kg⁻¹ diesel, and amended with nitrogen (15.98 atom% ¹⁵N) at 0, 250, 500, and 1000 mg kg⁻¹ to determine gross rates of nitrogen transformations during diesel biodegradation at varying soil water potentials. The observed water potential values were −0.20, −0.47, −0.85, and −1.50 MPa in the 0, 250, 500, and 1000 mg kg⁻¹ nitrogen treatments respectively. Highest microbial respiration occurred in the lowest nitrogen treatment suggesting an inhibitory osmotic effect from higher rates of nitrogen application. Microbial respiration rates of 185, 169, 131, and 116 mg O₂ kg⁻¹ soil day⁻¹ were observed in the 250, 500, control and 1000 mg kg⁻¹ nitrogen treatments, respectively. Gross nitrification was inversely related to water potential with rates of 0.2, 0.04, and 0.004 mg N kg⁻¹ soil day⁻¹ in the 250, 500, and 1000 mg kg⁻¹ nitrogen treatments, respectively. Reduction in water potential did not inhibit gross nitrogen immobilization or mineralization, with respective immobilization rates of 2.2, 1.8, and 1.8 mg N kg⁻¹ soil day⁻¹, and mineralization rates of 0.5, 0.3, and 0.3 mg N kg⁻¹ soil day⁻¹ in the 1000, 500, and 250 mg kg⁻¹ nitrogen treatments, respectively. Based on nitrogen transformation rates, the duration of fertilizer contribution to the inorganic nitrogen pool was estimated at 0.9, 1.9, and 3.2 years in the 250, 500, and 1000 mg kg⁻¹ nitrogen treatments, respectively. The estimation was conservative as ammonium fixation, gross nitrogen immobilization, and nitrification were considered losses of fertilizer with only gross mineralization of organic nitrogen contributing to the most active portion of the nitrogen pool.     

Enhanced biodegradation of diesel oil by a newly identified Rhodococcus baikonurensis EN3 in the presence of mycolic acid

AIMS: The aim of the present study was to isolate and characterize a bacterium, strain EN3, capable of using diesel oil as a major carbon and energy source, and to analyse the enhancement of diesel oil degradation by this organism using synthetic mycolic acid (2-hexyl-3-hydroxyldecanoic acid). METHOD AND RESULTS: An actinomycete with the ability to degrade diesel oil was isolated from oil contaminated soil and characterized. The strain had phenotypic properties consistent with its classification in the genus Rhodococcus showing a 16S rRNA gene similarity of 99.7% with Rhodococcus baikonurensis DSM 44587(T). The ability of the characterized strain to degrade diesel oil at various concentrations (1000, 5000, 10 000 and 20 000 mg l(-1)) was determined. The effect of synthetic mycolic acid on the biodegradation of diesel oil was investigated at the 20 000 mg l(-1) concentration; the surfactant was added to the flask cultures at three different concentrations (10, 50 and 100 mg l(-1)) and degradation followed over 7 days. Enhanced degradation was found at all three concentrations of the surfactant. In addition, the enhancement of diesel oil degradation by other surfactants was observed. CONCLUSIONS: The synthetic mycolic acid has potential for the remediation of petroleum-contaminated sites from both an economic and applied perspective as it can stimulate biodegradation at low concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed that the synthesized mycolic acid can be used for potential applications in the bioremediation industries, for example, in oil spill clean-up, diesel fuel remediation and biostimulation.     

Identification and Kinetic Characteristics of an Indigenous Diesel-degrading Gordonia alkanivorans Strain

Summary An indigenous strain Gordonia alkanivorans CC-JG39 was isolated from oil-contaminated sludge of a local gas station located in central Taiwan. The bacterial isolate was able to grow on diesel-containing Bushnell–Haas medium and also tolerate various chemical additives frequently used in petroleum products (e.g. BETX, methyl-tert-butyl ether, and naphthalene). Kinetics of diesel-limited cell growth and biodegradation of diesel followed a Monod-type model. The kinetic constants for cell growth (μ_max and K_S,G) were 0.158 h⁻¹ and 3196 mg/l, respectively, while those for biodegradation of diesel (v_{max, diesel} and K_S,D) were 3.59 mg/h/mg cell and 2874 mg/l, respectively. G. alkanivorans CC-JG39 produced extracellular surface-active material, leading to a low surface tension of nearly 33 mN/m. The CC-JG39 strain also possessed the ability to float towards the oil/water interface. These features might play some roles in enhancing the mass transfer efficiency between oil substrate and the bacterial cells. Therefore, G. alkanivorans CC-JG39 may have potential applications in bioremediation of oil pollution sites.     

Spatial and temporal impacts of a diesel fuel spill on stream invertebrates

1. We assessed the effects of a 26 500 L diesel fuel spill on the macroinvertebrate fauna of a small trout stream in central New York, U.S.A. To determine the spatial extent of the spill we sampled three locations (0.7, 5.0 and 11.8 km downstream of the spill), each containing a reference site (unaffected tributary) and an impact site (downstream of spill). Sampling was repeated four times over a 15‐month period to assess temporal recovery. 2. Immediately after the spill, invertebrate density at all three locations below the spill was significantly lower than reference density. Three months after the spill, density up to 5 km below the spill was still far lower (<100 individuals per sample) than reference density (800–1200 individuals per sample). A year after the spill, density was similar between reference and impact sites, suggesting that invertebrates had recovered numerically. 3. Taxonomic richness up to 5.0 km below the spill was less than half the reference taxonomic richness and this difference persisted for at least 3 months. Some significant differences between reference and impact sites were observed after 15 months, but these differences could not be attributed to the oil spill. 4. For at least 3 months following the spill, the site immediately downstream of the spill was dominated by Optioservus, a petrochemical‐tolerant riffle beetle. Twelve to 15 months after the spill, both the reference and impact sites near the spill were dominated numerically by the mayfly Ephemerella, but the degree of dominance was twice as large at the impact site. 5. We concluded that the diesel fuel spill significantly reduced the density of invertebrates (by 90%) and taxonomic richness (by 50%) at least 5.0 km downstream, but density recovered within a year. Throughout the study, however, the fauna immediately below the spill was species poor and significantly over‐represented by a single dominant taxon, suggesting that 15 months was not sufficient for full community recovery from the oil spill.