The biodegradation efficiency on diesel oil by two psychrotrophic Antarctic marine bacteria during a two-month-long experiment

Two psychrotrophic bacterial strains isolated from Antarctic seawaters were investigated for their capability to degrade commercial diesel oil. The efficiency of hydrocarbon utilization was studied at 4 and 20 degrees C over a period of two-months. Strains were cultured in a mineral liquid medium supplemented with diesel oil as the sole source of carbon and energy. The viable counts for the bacterial abundance estimation and the culture extractions for the subsequent gas-chromatographic analysis were carried out simultaneously. The biodegradation efficiency was higher at 20 degrees C than at 4 degrees C for both strains and the decrease in hydrocarbon concentrations reached more than 85% after 60 days of incubation at 20 degrees C. Our results suggest the possible exploitation of these two bacterial strains in future biotechnological processes, directly as field-released micro-organisms both in cold and temperate contaminated marine environments.     

Geochemical evolution during the cracking of crude oil into gas under different pressure systems

Two comparative simulation experiments(a normal atmospheric-pressure opening system and a 20 MPa closed system)were conducted to study the geochemical evolution of n-alkane,sterane,and terpane biomarkers in the process of oil cracking into gas under different pressures.With an initial experimental temperature set at 300°C,the temperature was increased to 650°C at a heating rate of 30°C/h.The products were tested every 50°C starting at 300°C,and a pressure of 20 MPa was achieved using a water column.The low-maturity crude oil sample was from the Paleogene system in the Dongying sag in eastern China.The threshold temperature obtained for the primary oil cracking process in both pressure systems was 450°C.Before the oil was cracked into gas,some components,including macromolecular n-alkanes,were cracked into medium-or small-sized n-alkanes.The secondary oil cracking of heavy hydrocarbon gases of C2–5to methane mainly occurred between 550°C to 650°C,and the parameters Ln(C1/C2)and Ln(C1/C3),as well as the dry coefficients,increased.Overpressure inhibited the oil cracking process.In the 20 MPa system,the oil conversion rate decreased,the temperature threshold for gas generation rose,and oil cracking was inhibited.Compared with the normal pressure system,high-carbon n-alkanes and other compounds in the 20 MPa pressure system were reserved.Furthermore,the parameters∑C21-/∑22+,Ln(C1/C2),and Ln(C1/C3),as well as the dry coefficients,decreased within the main temperature range.During secondary oil cracking(550°C to 600°C),the Ph/nC18and Pr/nC17decreased.High pressure influenced the evolution of the biomarkers Ts and Tm,C31homohopane,C29sterane,and their related maturity parameters to different extents during oil cracking under different temperature ranges.     

Screening of surfactants for harmful algal blooms mitigation

Screening experiments were conducted in order to find promising synthetic surfactants for harmful algal blooms (HABs) mitigation. The chemically synthesized surfactant cocamidopropyl betaine (CAPB) showed characteristics of relatively high inhibition efficiency, high biodegradability and low cost. The motility inhibition ratios of 10 mg/L CAPB on Cochlodinium polykrikoides and Alexandrium tamarense were about 60% after 5 min. The biodegradation test indicated that the half-life of CAPB in seawater was shorter than one day and 90% was biodegraded after five days under the initial concentration of 100 mg/L at 25 degrees C. Further cell lysis experiments revealed the selective lysis effect of CAPB on different HAB organisms. More than 90% of C. polykrikoides lysed at the concentration of 10 mg/L CAPB after 24 h and at 15 mg/L CAPB after 4 h, whereas the lysis effect of CAPB on A. tamarense was slight, no more than 10% after 2 h interaction with 50 mg/L CAPB. This research provided preliminary data for CAPB as a candidate in harmful algal blooms mitigation and pointed out unresolved problems for its practical application in the meantime.     

Effects of chitin on microbial emulsification, mineralization potential, and toxicity of bunker C fuel oil

Bunker C, one of the most frequently spilled petroleum products in the US, is difficult to remove from oiled surfaces and is relatively recalcitrant to biodegradation; therefore, emulsification and biodegradability must be optimized before bioremediation can be considered a viable treatment option. Sand from a freshly oiled beach near Dutch Harbor, Alaska, was incubated at 10 degrees C with nutrients (Bushnell-Haas (BH)) or nutrients with crab shell chitin (BH-C). BH-C amendment resulted in greater numbers of bunker C emulsifiers and greater mineralization potentials for hexadecane, phenanthrene, and fluorene than with BH only. Compared to BH alone, mineralization potentials for bunker C also were higher in BH-C, with an estimated 8% of fuel oil mineralized after 6 weeks. Microbially emulsified oil was more toxic than in uninoculated controls (p < 0.05) as measured by Microtox assays. However, toxicity was significantly lower in BH-C than BH after 4 and 6 weeks incubation (p < 0.05).     

Inhibition of Fertilization and Larval Metamorphosis of the Coral Acropora millepora (Ehrenberg, 1834) by Petroleum Products

Accidental oil spills from ships or rigs and inputs of effluent such as production formation water (PFW) are key perceived threats to tropical biota from industry activities. Scleractinian corals are an important functional component of tropical reefs and the abundance, diversity and resilience of coral communities can be used as an indicator of ecosystem health. In this paper, we report the effects of petroleum products, including water accommodated fractions (WAF) of crude oil, PFW and dispersant (Corexit 9527), on fertilization and larval metamorphosis of the widespread scleractinian coral, Acropora millepora (Ehrenberg, 1834) in laboratory-based assays. At 20% v/v PFW fertilization was inhibited by 25%. This concentration was equivalent 0.0721 mg l⁻¹ total hydrocarbon (THC). In contrast, larval metamorphosis was more sensitive to this effluent, with 98% metamorphosis inhibited at the same concentration. Crude oil WAF did not inhibit fertilization of gametes until dispersant was introduced. Dispersed oil was slightly more toxic to fertilization than dispersant alone, suggesting toxicity to that event may be additive. The minimum concentration of dispersed oil which inhibited fertilization was 0.0325 mg l⁻¹ THC. Larval metamorphosis was more sensitive than fertilization to crude oil. Although crude oil and dispersant inhibited larval metamorphosis individually, this toxicity was magnified when larvae were exposed to combinations of both. Crude oil inhibited metamorphosis at 0.0824 mg l⁻¹ THC and at 0.0325 mg l⁻¹ THC when dispersed in 10% v/v (dispersant/oil). Management of petroleum-related risks to spawning corals should consider not only the occurrence of the annual coral spawning event, but also the subsequent 1–3-week period during which most larval metamorphosis and recruitment occur.     

Experimental measurements of water content in crude oil emulsions by terahertz time-domain spectroscopy

We measured the water content （0.01% 0.25% w/w） in crude oil emulsions using terahertz time-domain spectroscopy （THz-TDS）. To improve the precision and range of the measurements, we used 1 and 10 mm thick quartz cells. The experiments were performed at 20 ℃ and the THz wave was transmitted vertically to the samples and detected on the other side. The experimental results suggest linear relation for the THz absorption coefficient and the water content of the crude oil emulsions in the observed range. The linear dependence facilitates high-precision measurements of the water content of crude oil. This suggests the potential of THz-TDS in determining the water concentration in crude oil and borehole fluid identification.     

Partitioning of semi-soluble organic compounds between the water phase and oil droplets in produced water

When selecting produced water treatment technologies, one should focus on reducing the major contributors to the total environmental impact. These are dispersed oil and semi-soluble hydrocarbons, alkylated phenols, and added chemicals. Experiments with produced water have been performed offshore on the Statoil operated platforms Gullfaks C and Statfjord B. These experiments were designed to find how much of the environmentally relevant compounds were dissolved in the water phase and not associated to the dispersed oil in the produced water. Results show that the distribution between the dispersed oil and the water phase varies highly for the different components groups. For example the concentration of PAHs and the C6-C9 alkylated phenols is strongly correlated to the content of dispersed oil. Therefore, the technologies enhancing the removal of dispersed oil have a higher potential for reducing the environmental impact of the produced water than previously considered.     

Consequences of linseed oil spills in salt marsh sediments

In a simulated spill in a salt marsh, linseed oil penetrated rapidly into the sediments at a rate of 10(-7) cm2 s(-1). The oil concentration remained unchanged for the first month after the spill, but 60% of the oil disappeared from the top 30 cm after a further month. The oil adsorbed to and accumulated in the muddy sediments (top 15 cm) leading to decreased sediment permeability, pH, Eh, abundance of plant roots and infauna and to the establishment of anoxic conditions. These changes accompanied transformations in the original fatty acid composition of the linseed oil, mainly associated with a decrease in 18 : 3omega3, an increase in the other fatty acids and the presence of 'new' fatty acids. A rapid increase in the abundance of heterotrophic aerobic and anaerobic bacteria and aerobic oil degrading bacteria, suggested that these micro-organisms degraded the oil. The role of the bacteria in oil degradation was confirmed in laboratory experiments where the fatty acids composition of the linseed oil underwent identical transformations to those obtained in the field. The degradation of linseed oil appears to be a sequential process initiated by aerobic and/or anaerobic bacteria and continued by sulphate reducing bacteria, which were unable to degrade the raw oil.     

Biodegradation of petroleum hydrocarbons at low temperature in the presence of the dispersant Corexit 9500

Our study examined the effects of Corexit 9500 and sediment on microbial mineralization of specific aliphatic and aromatic hydrocarbons found in crude oil. We also measured gross mineralization of crude oil, dispersed crude oil and dispersant by a marine microbial consortium in the absence of sediment. When provided as carbon sources, our consortium mineralized Corexit 9500 the most rapidly, followed by fresh oil, and finally weathered oil or dispersed oil. However, mineralization in short term assays favored particular components of crude oil (2-methyl-naphthalene > dodecane > phenanthrene > hexadecane > pyrene) and was not affected by addition of nutrients or sediment (high sand, low organic carbon). Adding dispersant inhibited hexadecane and phenanthrene mineralization but did not affect dodecane and 2-methyl-naphthalene mineralization. Thus, the effect of dispersant on biodegradation of a specific hydrocarbon was not predictable by class. The results were consistent for both high and low oiling experiments and for both fresh and weathered oil. Overall, our results indicate that environmental use of Corexit 9500 could result in either increases or decreases in the toxicity of residual oil through selective microbial mineralization of hydrocarbons.     

A case study on fuel oil contamination in a mangrove swamp in Hong Kong

Mangroves commonly found along tropical and subtropical coastlines are susceptible to oil pollution. In December 2000, around 500 1 m tall Kandelia candel saplings at the age of 3–5 years old located at the foreshore region of Sheung Pak Nai swamp, Hong Kong SAR, were found to be damaged by oil pollution. More than 80% of the saplings were either dead or washed away and leaving less than 5% healthy saplings with dense green leaves. Elevated concentrations of light n-alkanes (ranging from n-C₁₄ to n-C₂₀), pristane and phytane were recorded in surface sediments collected in December 2000. The ratio between light and total n-alkanes was 0.4. The total petroleum hydrocarbons (60–80 μg g⁻¹ TPH) and unresolved complex mixtures (60–70 μg g⁻¹ UCM) were higher than the background values of other mangrove sediments in Hong Kong, which were 40 and 20 μg g⁻¹, respectively. In certain root zone sediments, TPH concentrations were above 1000 μg g⁻¹. These results suggest that surface sediments in Sheung Pak Nai were contaminated by petroleum oil, most likely by illegal discharge of fuel oil which occurred between 1998 and 2002. One year later, in December 2001, unhealthy saplings had recovered and re-grown. The concentrations of TPH and UCM in sediments declined to around 40 μg g⁻¹, pristane and phytane dropped by 80%, and the ratio of light to total n-alkanes was 0.15, suggesting that residual oil in sediments was weathered leading to a remarkable recovery of the unhealthy saplings.     

Polycyclic aromatic hydrocarbon contamination and recovery characteristics in some organisms after the Nakhodka oil spill

Following the oil spill from the Russian tanker Nakhodka in 1997 in the Sea of Japan, polycyclic aromatic hydrocarbons (PAH) were monitored for three years in some molluscs from the Mikuni-cho shore in Japan. Total PAH concentrations in marine organisms except for spiny top shell, ranged from 5.3 to 32.7 ng/g wet weight, but no trends were evident. Total PAH concentration in spiny top shell (Turbo cornutus) was 44 ng/g w.w. in the first month after the oil spill. However, it rapidly decreased to less than 5.4 ng/g w.w. from the second month. Spiny top shell, which was exposed to dietary Nakhodka heavy fuel oil, concentrated benzo(a)pyrene to 17.1 ng/g w.w. after two weeks of exposure and then rapidly eliminated it during an elimination phase. These results suggest that spiny top shell accumulates PAHs because of their low ability to metabolize PAH, but it can excrete parent PAHs rapidly when removed from the source of contamination. Thus it is suitable as an indicator organism in monitoring oil contamination. It can also be inferred from these field and laboratory investigations that, in three years, organisms from the Mikuni-cho shore seem to have adequately recovered from the Nakhodka oil spill contamination.     

Long-term underground temperature measurements at Syowa Station, East Antarctica

Underground temperature measurements in two shallow boreholes have been carried out by the Japanese Antarctic Research Expedition at Syowa Station, East Antarctica from April, 1981 to January, 1985. Two quartz thermometers were installed in the first borehole at depths of 2 and 5 m and three were in the second one at depths of 1, 4 and 6.8 m. The mean underground temperatures in the first borehole were −8.181 and −8.843°C at depths of 2 and 5 m, and in the second one −8.242 and −8.220°C at depths of 4 and 6.8 m. As the mean air temperature at Syowa Station was −10.8°C, the underground temperature in the 2 −6.8 m depth range is about two degrees higher than the air temperature. The thermal diffusivities of the observation area are determined by the same principle of the Ångström method, using long-term underground temperature records. The thermal diffusivity around Syowa Station is established to be about two times larger than those of ordinary igneous and metamorphic rocks measured in the laboratory. The thermal conductivity of the drilled cores and surrounding outcropping rocks are also measured by the transient method with temperature conditions at +23°C and −20°C. The thermal conductivities measured in various samples at −20°C are about 7% larger than those at +23°C. Thes tendency is consistent with the results of holocrystalline rock experiments.     

Determining the dispersibility of South Louisiana crude oil by eight oil dispersant products listed on the NCP Product Schedule

We recently conducted a laboratory study to measure the dispersion effectiveness of eight dispersants currently listed on the National Contingency Plan Product Schedule. Results are useful in determining how many commercial dispersant products would have been effective for use on South Louisiana crude oil in the Deepwater Horizon oil spill. The test used was a modification of the Baffled Flask Test (BFT), which is being proposed to replace the current Swirling Flask Test (SFT). The modifications of the BFT in this study included use of one oil rather than two, increasing replication from 4 runs to 6, and testing at two temperatures, 5 °C and 25 °C. Results indicated that temperature was not as critical a variable as the literature suggested, likely because of the low viscosity and light weight of the SLC. Of the eight dispersants tested, only three gave satisfactory results in the laboratory flasks at both temperatures.     

Oil droplet interaction with suspended sediment in the seawater column: Influence of physical parameters and chemical dispersants

The interaction of dispersed oil droplets with large diameter suspended particulate materials (SPM) has been little studied. In the current study, particle size, oil characteristics and chemical dispersant significantly influence the adsorption of oil droplets to SPM in seawater. Sediments with a smaller particulate size (clay) approaching that of the oil droplets (2–20 μm) adsorbed more oil per gram than sediments with large particle size (sand). Heavier, more polar oils with a high asphaltene content adsorbed more efficiently to SPM than lighter, less polar oils. A decrease in the smaller, more water soluble oil components in the sediment adsorbed oil was observed for all oil types. Addition of chemical dispersant decreased the adsorption of oil droplets to suspended carbonate sand in an exponential-like manner. No change in the relative distribution of compounds adsorbed to the sediment was observed, indicating dispersants do not alter the dissolution of compounds from oil droplets.     

Gladstone, Australia Field Studies: Weathering and Degradation of Hydrocarbons in Oiled Mangrove and Salt Marsh Sediments With and Without the Application of an Experimental Bioremediation Protocol

This field study was a combined chemical and biological investigation of the relative rates of weathering and biodegradation of oil spilled in sediments and testing the influence of a bioremediation protocol. The aim of the chemistry work presented here was to determine whether the bioremediation protocol affected the rate of penetration, dissipation or long-term retention of a medium range crude oil (Gippsland) and a Bunker C oil stranded in tropical Rhizophora sp. mangrove and Halosarcia sp. salt marsh environments. Permission for the planned oil spills was granted in the Port Authority area of Gladstone, Queensland (Australia). Sediment cores from three replicate plots of each treatment for mangroves and four replicate plots for the salt marsh (oil only and oil plus bioremediation) were analysed for total hydrocarbons (THC) and for individual alkane markers using gas chromatography with flame ionization detection (GC–FID). Sediments were collected at day 2, then 1, 2, 5 or 6 and 12 or 13 months post-spill for mangroves and day 2, 1, 3 and 9 months post-spill for salt marshes. Over this time, hydrocarbons in all of the oil treated plots decreased exponentially. There was no statistical difference in initial oil concentrations, penetration of oil to depth, or in the rates of oil dissipation between untreated oil and bioremediated oil in the mangrove plots. The salt marsh plots treated with the waxy Gippsland oil showed a faster rate of biodegradation of the oil in the bioremediated plots. In this case only, the degradation rate significantly impacted the mass balance of remaining oil. The Bunker C oil contained only minor amounts of highly degradable n-alkanes and bioremediation did not significantly impact its rate of loss in the salt marsh sediments. At the end of each experiment, there were still n-alkanes visible in the gas chromatograms of residual oils. Thus it was concluded that there was unlikely to be any change in the stable internal biomarkers of the oils over this time period. The predominant removal processes in both habitats were evaporation and dissolution, with a lag-phase of 1–2 months before the start of microbial degradation.     

Effects of temperature and wave conditions on chemical dispersion efficacy of heavy fuel oil in an experimental flow-through wave tank

The effectiveness of chemical dispersants (Corexit 9500 and SPC 1000) on heavy fuel oil (IFO180 as test oil) has been evaluated under different wave conditions in a flow-through wave tank. The dispersant effectiveness was determined by measuring oil concentrations and droplet size distributions. An analysis of covariance (ANCOVA) model indicated that wave type and temperature significantly (p < 0.05) affected the dynamic dispersant effectiveness (DDE). At higher temperatures (16 °C), the test IFO180 was effectively dispersed under breaking waves with a DDE of 90% and 50% for Corexit 9500 and SPC 1000, respectively. The dispersion was ineffective under breaking waves at lower temperature (10 °C), and under regular wave conditions at all temperatures (10-17 °C), with DDE < 15%. Effective chemical dispersion was associated with formation of smaller droplets (with volumetric mean diameters or VMD ? 200 μm), whereas ineffective dispersion produced large oil droplets (with VMD ? 400 μm).     

Toxicogenomic analysis of immune system-related genes in Japanese flounder (Paralichthys olivaceus) exposed to heavy oil

Heavy oil contamination is one of the most important environmental issues. Toxicities of polycyclic aromatic hydrocarbons (PAHs), including immune toxicities, are well characterized, however, the immune toxic effects of heavy oil, as a complex mixture of PAHs, have not been investigated. In the present study, we selected Japanese flounder (Paralichthys olivaceus) as a model organism, and observed alteration of immune function by the exposure to heavy oil. To analyze the expression profiles of immune system-related genes, we selected 309 cDNAs from our flounder EST library, and spotted them on a glass slide. Using this cDNA array, alteration of gene expression profiles was analyzed in the kidneys of flounders exposed to heavy oil. Six Japanese flounders (mean body weight: 197 g) were acclimated to laboratory conditions at 19–20 °C. Three fish were exposed to heavy oil C (bunker C) at a concentration of 3.8 g/L for 3 days, and the others were kept in seawater without heavy oil and used as the control. After the exposure period, the fish were transferred into control seawater and maintained for 4 days, and then they were dissected and their kidneys were removed. Total RNA was extracted from the kidney samples to use in gene expression analyses. The microarray detected alteration of immune system-related genes in the kidneys of heavy oil-exposed flounders, including down-regulation of immunoglobulin light chain, CD45, major histocompatibility complex class II antigens and macrophage colony-stimulating factor precursor, and up-regulation of interleukin-8 and lysozyme. These results suggest that pathogen resistance may be weakened in heavy oil-exposed fish, causing a subsequent bacterial infection, and then proinflammatory genes may be induced as a defensive response against the infection. Additionally, we found candidate genes for use as biomarkers of heavy oil exposure, such as N-myc downstream regulated gene 1 and heat shock cognate 71 kDa proteins.     

Effects of chronic exposure to dispersed oil on selected reproductive processes in adult blue mussels (Mytilus edulis) and the consequences for the early life stages of their larvae

Mussels (Mytilus edulis) were continuously exposed to dispersed crude oil (0.015-0.25 mg/l) for 7 months covering the whole gamete development cycle. After 1 month exposure to 0.25 mg oil/l, the level of alkali-labile phosphates (ALP) and the volume density of atretic oocytes in female gonads were higher than those in the gonads of control females, indicating that oil affected the level of vitellogenin-like proteins and gamete development. Spawning of mussels was induced after 7 months oil exposure. Parental oil exposure did not affect subsequent fertilization success in clean seawater but this was reduced in 0.25 mg oil/l. Parental exposure to 0.25 mg oil/l caused both slow development and a higher percentage of abnormalities in D-shell larvae 2 days post-fertilization; reduced growth 7 days post-fertilization. These effects were greatly enhanced when larval stages were maintained at 0.25 mg oil/l. Similar studies are warranted for risk assessment prognosis.     

Weathering of an Arctic oil spill over 20 years: the BIOS experiment revisited. Baffin Island Oil Spill

A small experimental oil spill was conducted on the northern tip of Baffin Island Nunavut, Canada (72 degrees 31' N, 79 degrees 50' W) in August 1981, and the natural weathering of the oil has been followed by periodic visits. This paper reports on the chemical composition of oil collected in August 2001. The vast majority of the initial oil has gone, but small patches remain. Some samples remain essentially unaltered despite their 20 years of exposure to the elements, while others show that biodegradation and photooxidation can play important roles in removing the majority of the components of the oil. Using 17alpha(H)21beta(H)hopane as a conserved marker within the oil, we show that the most biodegraded sample has lost more than 87% of the hydrocarbons initially present, while another has lost a substantial proportion of its initial chrysene and alkylated congeners. Potential explanations for the different weathering patterns seen in samples collected from this small site are briefly discussed.     

Sorption of heavy oil onto Jiaozhou Bay sediment

The sorption of heavy oil onto sediment collected from Jiaozhou Bay was studied in a series of kinetic and equilibrium experiments using NaCl solutions. The effects of temperature, salinity, and pH of the medium on sorption behavior were investigated.Sorption equilibrium of the heavy oil and sediment was established within 60 min. The process was shown to follow a pseudo-second-order kinetic rate model. The sorption rate decreased with increasing initial heavy oil concentration in the solution. Batch equilibrium experiments showed that the sorption isotherm could be described by the Freundlich model. The standard free energy change and enthalpy change at the temperatures studied (283, 288, 293, and 298 K) were negative. These findings indicated that the process was spontaneous and exothermic. Salinity, pH and temperature influenced sorption performance. Sorption was favored by higher concentrations of NaCl, by lower pH values and by lower temperatures.