Jee-Hyun Jung

Fingerprint and weathering characteristics of stranded oils after the Hebei Spirit oil spill.

After the Hebei Spirit oil spill in December 2007, mixtures of three types of Middle East crude oil were stranded along 375 km of coastline in Western Korea. Stranded oils were monitored for their identity and weathering status in 19 stations in three provinces. The results obtained using a weathering model indicated that evaporation would be a dominant weathering process immediately after the spill and the sequential changes of chemical composition in the field verified this prediction positively. In the early stages of weathering, the half-life of spilled oil was calculated to be 2.6 months. Tiered fingerprinting approaches identified background contamination and confirmed the identity of the stranded oils with the spill source. Double ratios using alkylated phenanthrenes and dibenzothiophenes in samples after the spill clearly reveal the impact of weathering on oil. However, to derive defensible fingerprinting for source identification and allocation, recalcitrant biomarkers are extremely useful. Weathering status of the stranded oils was evaluated using composition profiles of saturated hydrocarbons, polycyclic aromatic hydrocarbons and various weathering indices. Most samples collected 8 months after the spill were categorized in either the advanced or extreme weathering states. Gradual increase in toxic components in the residual oil through weathering emphasizes the need for adaptive ecotoxicological approaches.

Hebei Spirit oil spill monitored on site by fluorometric detection of residual oil in coastal waters off Taean, Korea.

The spatiotemporal distributions of dissolved and/or dispersed oil in seawater and pore water were monitored on site by fluorometric detection method after the Hebei Spirit oil spill. The oil concentrations in intertidal seawater, 15 days after the spill, were as high as 16,600 microg/L and appeared to decrease below the Korean marine water quality standard of 10 microg/L at most sites 10 months after the spill. Fluorometric detection of oil in pore water was introduced to eliminate the effects of grain size for the quantification of oil in sediments and to better explain spatial and temporal distribution of oil pollution at sandy beaches. The fluorescence detection method was compared with the conventional laboratory technique of total petroleum hydrocarbon analysis using gas chromatography. The method of fluorescence detection of oil was capable of generating results much faster and more cost-effectively than the traditional GC technique.

Biomarker responses in pelagic and benthic fish over 1 year following the Hebei Spirit oil spill (Taean, Korea)

After the Hebei Spirit oil spill incident (7th December, 2007) in the west coast of Korea, contamination of biliary PAH metabolite and hepatic biomarkers in a pelagic and a benthic fish was monitored for 1 year. Concentrations of 16 PAHs and alkylated PAHs in fish muscle were highest (22.0 ng/g d.w. for 16 PAHs and 284 ng/g d.w. for alkylated PAHs) at 5 days after the spill and then decreased rapidly to background levels at 11 months after the spill. Fish from the oiled site had elevated biliary PAH metabolite concentrations immediately after the spill; these declined steadily in both species, but were still above reference site concentrations 2 months after the spill. Oiled-site fish showed hepatic CYP 1A induction whose trend closely followed those of biliary PAH metabolite concentrations, implying continuous exposure to PAHs. Brain acetylcholinesterase activity was not related to oil exposure.

Geologically distinct crude oils cause a common cardiotoxicity syndrome in developing zebrafish.

Crude oils from different geological formations vary in composition, yet most crude oils contain a polycyclic aromatic hydrocarbon (PAH) fraction that would be expected to produce cardiotoxic effects in developing fish. To determine whether different crude oils or PAH compositions produce common or distinct effects, we used zebrafish embryos to directly compare two crude oils at different states of weathering. Iranian heavy crude oil (IHCO) spilled in the Yellow Sea following the 2007 Hebei Spirit accident was compared to the intensively studied Alaska North Slope crude oil (ANSCO) using two different exposure methods, water-accommodated fractions containing dispersed oil microdroplets and oiled gravel effluent. Overall, both crude oils produced a largely overlapping suite of defects, marked by the well-known effects of PAH exposure on cardiac function. Specific cardiotoxicity phenotypes were nearly identical between the two oils, including impacts on ventricular contractility and looping of the cardiac chambers. However, with increased weathering, tissue-specific patterns of aryl hydrocarbon receptor (AHR) activation in the heart changed, with myocardial AHR activation evident when alkyl-PAHs dominated the mixture. Our findings suggest that mechanisms of cardiotoxicity may shift from a predominantly AHR-independent mode during early weathering to a multiple pathway or synergistic mode with prolonged weathering and increased proportions of dissolved alkyl-PAHs. Despite continued need for comparisons of crude oils from different sources, the results here indicate that the body of knowledge already acquired from studies of ANSCO is directly relevant to understanding the impacts of other crude oil spills on the early life history stages of fish.

Petroleum hydrocarbon contaminations in the intertidal seawater after the Hebei Spirit oil spill ― Effect of tidal cycle on the TPH concentrations and the chromatographic characterization of seawater extracts

In December 2007, the oil tanker Hebei Spirit released approximately 12,547,000 L of crude oil off the west coast of Korea, impacting more than 375 km of coastline. The seawater TPH concentrations immediately after the spill ranged from 1.5 to 7310 μg L⁻¹, with an average of 732 μg L⁻¹. The concentrations appeared to decrease drastically to 2.0-224 μg L⁻¹ in one month after the spill. The TPH concentrations in seawater fluctuated with time thereafter because of the remobilization of oil by continuing shoreline cleanup activities and subsequent wave/tidal actions. Seawater TPH concentrations were much higher during high tide than during low tide due to the resuspension of stranded oil. The variation of TPH levels in seawater also matched the spring-neap tidal cycle in the study areas for the first three weeks of the study. Comparisons of the gas chromatograms of the seawater with the water accommodated fraction and the cargo oil indicated that seawater samples were contaminated mainly by the dispersed droplets of spilled oil. One year of monitoring revealed that the oil content in seawater had clearly decreased at most sites, although some regional fluctuations of oil contamination were noted until June 2008.

Evaluation of biomarker potential of cytochrome P450 1A (CYP1A) gene in the marine medaka, Oryzias melastigma exposed to water-accommodated fractions (WAFs) of Iranian crude oil

CYP1A is involved in the metabolism of diverse chemicals, including polycyclic aromatic hydrocarbons and alkylated-PAHs, as a first line of detoxification mechanism. First, we identified and characterized the CYP1A gene from the marine medaka, Oryzias melastigma. O. melastigma CYP1A (Om-CYP1A) showed a high similarity of motifs/domains compared to those of vertebrates in their amino acid sequences. To check whether the Om-CYP1A would be inducible, we tested two strong CYP1A inducers, β-naphthoflavone (β-NF) and benzo[α]pyrene (B[α]P), and observed concentration-dependent transient expression on transcripts of Om-CYP1A for 96 h over a wide range of concentrations. Om-CYP1A mRNA level was significantly increased in exposure to different concentrations of β-NF and B[α]P, and its expression was highly transcribed within 12 h upon the exposure to low concentrations of both chemicals. Inducible transcript profiles revealed that Om-CYP1A would be associated with the toxicant metabolism via AhREs/DREs/XREs in its promoter region. To uncover the effects of the water-accommodated fraction (WAF) of crude oil on transcripts of Om-CYP1A, we measured mRNA expression of Om-CYP1A towards different concentrations of WAF for 24h. As a result, WAF exposure significantly increased Om-CYP1A transcripts at all concentrations as well as during time-course experiments for 96 h. In this paper, we demonstrated that WAF would trigger up-regulation of the CYP1A gene that would be associated with the initiation of the cellular defense systems. This finding provides a better understanding of the molecular mechanism of cellular protection particularly that involved in the WAF-mediated cellular response in O. melastigma.

Spatial variability of biochemical responses in resident fish after the M/V Hebei Spirit Oil Spill (Taean, Korea)

This study describes the spatial variation and the duration of the impacts from the Hebei Spirit oil spill using specific biochemical indices in resident benthic fish. Enzymatic activities and biliary PAHs metabolites were higher at the site closer to the spill area in four months after spill incident. Regarding our results of detoxification response, markers of Phase I followed a similar trend in accordance with levels of biliary metabolites, while markers of phase II and GST appeared relatively unchanged.

Molecular characterization, expression, and functional analysis of two thioredoxins in the black rockfish (Sebastes schlegelii).

Thioredoxins (TRxs) are a family of small evolutionarily conserved proteins that are essential for the maintenance of cellular homeostasis. Two TRx homologue cDNAs were isolated from a black rockfish concanavalin A (Con A)/phorbol myristate acetate (PMA)-stimulated leucocyte cDNA library and named BrTPx1-1 and BrTPx1-2. As compared with other known TRx peptide sequences, the most conserved regions of both BrTRx1-1 and BrTRx1-2 peptides were found to be the redox-active site Trp-Cys-X-X-Cys (WCXXC). The TRx present in most species is a TRx1-2 protein with a Cys-Pro-Gly-Cys (CPGC) active site. However, in the larger 13 kDa BrTRx1-1 protein, a Cys-Pro-Pro-Cys (CPPC) active site was identified. Here, we report the identification of a new member of the TRx protein family from the teleost black rockfish, which defines a new subclass of 13-kDa TRx1-1 proteins. Phylogenetic analysis indicated that both BrTRx1-1 and BrTRx1-2 were grouped with other vertebrate TRx1 peptides. BrTRx1-1 expression was strongly induced in peripheral blood leucocytes (PBLs) 12-24 h following Con A/PMA stimulation, with peak expression at 24 h post-stimulation. BrTRx1-2 was induced in PBLs after stimulation with lipopolysaccharide (LPS), Con A/PMA, or poly I:C at 24 h. The BrTRx1-1 gene was predominantly expressed in the liver and gills, while BrTRx1-2 was expressed in PBLs and gills. After treatment with a high concentration (10 μg/mL) of rBrTRx1-1 or rBrTRx1-2, kidney leucocytes exhibited increased cell proliferation and viability under oxidative stress.

Combined Effects of Iranian Heavy Crude Oil and Bacterial Challenge ( Streptococcus iniae ) on Biotransformation and Innate Immune Responses in Rockfish ( Sebastes schlegeli )

To clarify the effects of spilled crude oil on fish bacterial disease resistance, rockfish (Sebastes schlegeli) were exposed to Iranian Heavy crude oil (IHCO) and Streptomyces iniae in combination. Hepatic biotransformation enzymes (ethoxyresorufin O-de-ethylase, glutathione-S-transferase) and plasma biochemical parameters (glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and glucose) in fish exposed to IHCO were not significantly different from those in unexposed fish. The level of biliary 1-OH-pyrene and cytochrome P4501A mRNA expression increased in a dose-dependent manner with IHCO exposure. The interferon stimulated gene 15, interleukin-1beta and cathepsin L were increased significantly in the liver in IHCO-exposed fish, but not dose-dependently, but the granulocyte colony stimulating factor was not related to IHCO exposure. The percentage mortality in fish following a single exposure to S. iniae was positively correlated with IHCO exposure concentration. We concluded that IHCO exposure exacerbates fish mortality following environmental bacterial infection.

Origins of suspended particulate matter based on sterol distribution in low salinity water mass observed in the offshore East China Sea.

The molecular composition and distribution of sterols were investigated in the East China Sea to identify the origins of suspended particulate matter (SPM) in offshore waters influenced by Changjiang River Diluted Water (CRDW). Total sterol concentrations ranged from 3200 to 31,900pgL(-1) and 663 to 5690pgL(-1) in the particulate and dissolved phases, respectively. Marine sterols dominated representing 71% and 66% in the particulate and dissolved phases, respectively. Typical sewage markers, such as coprostanol, were usually absent at ~250km offshore. However, sterols from allochthonous terrestrial plants were still detected at these sites. A negative relationship was observed between salinity and concentrations of terrestrial sterols in SPM, suggesting that significant amounts of terrestrial particulate matter traveled long distance offshore in the East China Sea, and the Changjiang River Diluted Water (CRDW) was an effective carrier of land-derived particulate organic matter to the offshore East China Sea.