Cytochrome P4501A expression,chemical contaminants and histopathology in roach,goby and sturgeon and chemical contaminants in sediments from the Caspian Sea,Lake Balkhash and the Ily River Delta,Kazakhstan

Roach, goby and sturgeon were examined for cytochrome P4501A (CYP1A) expression and histopathology, in relation to contaminant burdens in fish and sediment. Gradients of induction of CYP1A were observed. Roach from the Ural and Ily River Deltas and roach and goby from the two stations nearest the Caspian Sea oil fields displayed higher levels of CYP1A expression in several organs than was observed in fish from further offshore. Great sturgeon and Russian sturgeon showed higher levels of CYP1A expression than was seen in starred sturgeon and gobies in the Ural delta. No fish showed evidence of contaminant-related histopathologies in the organs examined, despite the elevated CYP1A levels. Low levels of polychlorinated biphenyls and elevated levels of inshore and riverine petroleum hydrocarbons from these habitats suggest that this ongoing hydrocarbon exposure, and that from natural sources and long-term oil exploration on the Northeastern Caspian shore, contributed to the CYP1A induction observed.     

Summary results from a pilot study conducted around an oil production platform on the Northwest Shelf of Australia

The Australian Institute of Marine Science (AIMS) conducted a pilot study around the Harriet A oil production platform on the Northwest Shelf of Australia. We evaluated hepatic ethoxyresorufin-O-deethylase (EROD) activity, fluorescent aromatic compounds (FACs) in bile and immunodetection of CYP1A-like proteins in two Australian tropical fish species, Gold-Spotted Trevally (Carangoides fulvoguttatus) and Bar-Cheeked Coral Trout (Plectropomus maculatus) to assess exposure to petroleum hydrocarbons associated with produced formation water (PFW). Additionally, the incidence of hydrocarbon-degrading bacteria isolated from the liver and bile of all fish captured was examined. Low EROD activity was found in both species, with EROD activity in C. fulvoguttatus showing significant site differences. FACs and CYP1A protein levels in C. fulvoguttatus showed a clear trend in hydrocarbon exposure consistent with hydrocarbon chemistry data: Harriet A>Harriet C>reference site. P. maculatus showed elevated levels of FACs at Harriet A as compared to the reference site and demonstrated detectable levels of CYP1A-like proteins at these two sites. Hydrocarbon-degrading bacteria were found in the liver and bile of both species, yet there was no correlation by sites. Our results demonstrate that C. fulvoguttatus and P. maculatus have potential as indicator species for assessing the effects from exposure to petroleum hydrocarbons. Both FACs and CYP1A are providing warning signs that there is potential for biological effects on fish populations exposed to PFW around the Harriet A production platform.     

Expression of superoxide dismutase (SOD) genes from the copper-exposed polychaete, Neanthes succinea

To evaluate the superoxide dismutase (SOD) genes as a potential indicator of environmental stress, we cloned and sequenced the full-length cDNA of a Cu/Zn-SOD and Mn-SOD genes from the marine polychaete, Neanthes succinea. Ns-Cu/Zn- and Mn-SOD expression was measured using quantitative real-time RT-PCR in response to the oxidative stress induced by copper in the form of CuCl2. Ns-Cu/Zn-SOD expression increased in a dose dependent manner but the increase of Mn-SOD expression was evident only at a high concentration of copper. Particularly, Ns-Mn-SOD dramatically increased in the 72 μg/L CuCl2-exposed groups, indicating that both Ns-SOD genes were induced by copper exposure. These results suggest that Ns-Cu/Zn- and Ns-Mn-SOD would play important roles in the detoxification of cellular oxidative stresses in the polychaete, N. succinea. Furthermore, as part of field work, the marine polychaete, N. succineaCu/Zn-SOD mRNA collected from the polluted area was accordingly upregulated in expression but not for NS-Mn-SOD.     

Molecular cloning and mRNA expression of cytochrome P4501A1 and 1A2 in the liver of common minke whales (Balaenoptera acutorostrata)

This study presents full-length cDNA sequences of CYP1A1 and 1A2, in common minke whale (Balaenoptera acutorostrata) from the North Pacific. Both CYP1A1 and CYP1A2 cDNAs had an open reading frame of 516 amino acid residues, and predicted molecular masses were 58.3 kDa and 58.1 kDa, respectively. The deduced full-length amino acid sequence of CYP1A1 revealed higher identities with those of sheep (86%) and pig (87%), and that of CYP1A2 was most closely related to human (82%) and monkey CYP1A2 (82%) among species from which CYP1A2 has been isolated so far. Differences in certain conserved and functional amino acid residues of CYP1A1 and 1A2 between common minke whale and other mammalian species indicate the possibility of their specific metabolic function. Concentrations of organochlorine compounds (OCs) including PCBs and DDTs analyzed in common minke whale liver showed no significant correlation with hepatic mRNA expression levels of CYP1A1 and CYP1A2, indicating no induction of these enzymes by such OCs.     

Molecular cloning and mRNA expression of cytochrome P4501A1 and 1A2 in the liver of common minke whales (Balaenoptera acutorostrata)

This study presents full-length cDNA sequences of CYP1A1 and 1A2, in common minke whale (Balaenoptera acutorostrata) from the North Pacific. Both CYP1A1 and CYP1A2 cDNAs had an open reading frame of 516 amino acid residues, and predicted molecular masses were 58.3 kDa and 58.1 kDa, respectively. The deduced full-length amino acid sequence of CYP1A1 revealed higher identities with those of sheep (86%) and pig (87%), and that of CYP1A2 was most closely related to human (82%) and monkey CYP1A2 (82%) among species from which CYP1A2 has been isolated so far. Differences in certain conserved and functional amino acid residues of CYP1A1 and 1A2 between common minke whale and other mammalian species indicate the possibility of their specific metabolic function. Concentrations of organochlorine compounds (OCs) including PCBs and DDTs analyzed in common minke whale liver showed no significant correlation with hepatic mRNA expression levels of CYP1A1 and CYP1A2, indicating no induction of these enzymes by such OCs.     

Molecular cloning and beta-naphthoflavone-induced expression of a cytochrome P450 1A (CYP1A) gene from an anadromous river pufferfish, Takifugu obscurus

In recent years, there has been a decline in the wild populations of river pufferfish, Takifugu obscurus. Besides overexploitation for commercial purposes, environmental pollution is believed to have contributed to its decline. However, almost no information exists about genes involved in metabolism of xenobiotics by this species. Nevertheless, there is interest in fugu fishes, since they possess the smallest genome among vertebrates. We cloned and characterized the full-length cDNA sequence of a cytochrome P450 1A (CYP1A) gene from T. obscurus. Phylogenic relationship of T. obscurus CYP1A was also compared to other fish species. The tissue distribution and time-dependant induction of CYP1A mRNA were studied by real-time PCR in T. obscurus exposed to an aryl hydrocarbon receptor (Ahr) agonist, beta-naphthoflavone (BNF). The greatest basal expression in livers of control as well as BNF-treated individuals. However, brain, gill, gonad, intestine, and kidney also expressed CYP1A. Muscles expressed the least CYP1A. The results of the time-course study revealed induction in brain and gills after 6h and at 12h in most tissues. Except for gills, all other organs retained induced expression of CYP1A mRNA up to 96h.     

Benzo[a]pyrene modulates the biotransformation,DNA damage and cortisol level of red sea bream challenged with lipopolysaccharide

In animals, biotransformation and the immune system interact with each other, however, knowledge of the toxic mechanism of benzo[a]pyrene (BaP) on these two systems is not well known. The present study investigated the toxic effects of BaP on the biotransformation system, cortisol level and DNA integrity of red sea bream (Pagrus major). The results showed that cortisol level was induced under the challenge of lipopolysaccharide (LPS). Short-term exposure (96 h) of BaP at environmental concentration significantly increased the cortisol level, hepatic EROD activity and CYP1A1 mRNA expression. When P. major was exposed to BaP for 14 d followed by LPS challenge this increased the cortisol level, EROD activity and hepatic DNA damage except CYP1A1 mRNA expression. Combined with our previous data, which showed that BaP exposure can modulate the immunologic response in P. major challenged with LPS, a hypothetical adverse outcome pathway of BaP on fish was suggested.     

Molecular cloning and β-naphthoflavone-induced expression of a cytochrome P450 1A (CYP1A) gene from an anadromous river pufferfish, Takifugu obscurus

In recent years, there has been a decline in the wild populations of river pufferfish, Takifugu obscurus. Besides overexploitation for commercial purposes, environmental pollution is believed to have contributed to its decline. However, almost no information exists about genes involved in metabolism of xenobiotics by this species. Nevertheless, there is interest in fugu fishes, since they possess the smallest genome among vertebrates. We cloned and characterized the full-length cDNA sequence of a cytochrome P450 1A (CYP1A) gene from T. obscurus. Phylogenic relationship of T. obscurus CYP1A was also compared to other fish species. The tissue distribution and time-dependant induction of CYP1A mRNA were studied by real-time PCR in T. obscurus exposed to an aryl hydrocarbon receptor (Ahr) agonist, β-naphthoflavone (BNF). The greatest basal expression in livers of control as well as BNF-treated individuals. However, brain, gill, gonad, intestine, and kidney also expressed CYP1A. Muscles expressed the least CYP1A. The results of the time-course study revealed induction in brain and gills after 6 h and at 12 h in most tissues. Except for gills, all other organs retained induced expression of CYP1A mRNA up to 96 h.     

Mutation analysis of ras gene in the liver of European eel (Anguilla anguilla L.) exposed to benzo[a]pyrene

Ras is regarded as one of the most important genes involved in carcinogenesis. Such genes have been characterised in several fish species and the presence of ras mutations have already been described in fish populations from hydrocarbon contaminated areas and following experimental exposure to specific contaminants. The aims of this study were to evaluate the DNA integrity by comet assay, to isolate the normal ras gene of Anguilla anguilla and analyse for the presence of ras gene mutations or changes in gene expression levels induced after one month of benzo[a]pyrene (BaP) experimental exposure. The A. anguilla ras cDNA isolated revealed a 189 amino acid protein and alignment with other vertebrate ras proteins revealed conservation of functionally important regions. Following experimental exposure to BaP, an increase in DNA damage was found by comet assay. However, no point mutations or changes in ras gene expression levels were detected when compared to control samples. In contrast to the majority of fish ras gene sequences, a high degree of polymorphic variation was detected in the A. anguilla ras gene.     

Vapor Intrusion Screening at Petroleum UST Sites

Detailed site investigations to assess potential inhalation exposure and risk to human health associated with the migration of petroleum hydrocarbon vapors from the subsurface to indoor air are frequently undertaken at leaking underground storage tank (UST) sites, yet documented occurrences of petroleum vapor intrusion are extremely rare. Additional assessments are largely driven by low screening‐level concentrations derived from vapor transport modeling that does not consider biodegradation. To address this issue, screening criteria were developed from soil‐gas measurements at hundreds of petroleum UST sites spanning a range of environmental conditions, geographic regions, and a 16‐year time period (1995 to 2011). The data were evaluated to define vertical separation (screening) distances from the source, beyond which, the potential for vapor intrusion can be considered negligible. The screening distances were derived explicitly from benzene data using specified soil‐gas screening levels of 30, 50, and 100 µg/m³ and nonparametric Kaplan‐Meier statistics. Results indicate that more than 95% of benzene concentrations in soil gas are ≤30 µg/m³ at any distance above a dissolved‐phase hydrocarbon source. Dissolved‐phase petroleum hydrocarbon sources are therefore unlikely to pose a risk for vapor intrusion unless groundwater (including capillary fringe) comes in contact with a building foundation. For light nonaqueous‐phase liquid (LNAPL) hydrocarbon sources, more than 95% of benzene concentrations in soil gas are ≤30 µg/m³ for vertical screening distances of 13 ft (4 m) or greater. The screening distances derived from this analysis are markedly different from 30 to 100 ft (10 to 30 m) vertical distances commonly found cited in regulatory guidance, even with specific allowances to account for uncertainty in the hydrocarbon source depth or location. Consideration of these screening distances in vapor intrusion guidance would help eliminate unnecessary site characterization at petroleum UST sites and allow more effective and sustainable use of limited resources.     

Expression of superoxide dismutase (SOD) genes from the copper-exposed polychaete,Neanthes succinea

To evaluate the superoxide dismutase (SOD) genes as a potential indicator of environmental stress, we cloned and sequenced the full-length cDNA of a Cu/Zn-SOD and Mn-SOD genes from the marine polychaete, Neanthes succinea. Ns-Cu/Zn- and Mn-SOD expression was measured using quantitative real-time RT-PCR in response to the oxidative stress induced by copper in the form of CuCl₂. Ns-Cu/Zn-SOD expression increased in a dose dependent manner but the increase of Mn-SOD expression was evident only at a high concentration of copper. Particularly, Ns-Mn-SOD dramatically increased in the 72 μg/L CuCl₂-exposed groups, indicating that both Ns-SOD genes were induced by copper exposure. These results suggest that Ns-Cu/Zn- and Ns-Mn-SOD would play important roles in the detoxification of cellular oxidative stresses in the polychaete, N. succinea. Furthermore, as part of field work, the marine polychaete, N. succinea Cu/Zn-SOD mRNA collected from the polluted area was accordingly upregulated in expression but not for NS-Mn-SOD.     

Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.     

Effect of zinc sacrificial anode degradation on the defence system of the Pacific oyster, Crassostrea gigas: Chronic and acute exposures

Two types of exposures were performed to assess the effects of zinc released from sacrificial anode degradation: a chronic exposure, in which oysters were exposed to 0.53±0.04mgZnL(-1) for 10weeks, and an acute exposure, where oysters were exposed to 10.2±1.2mgZnL(-1) for 1week. At the end of the acute exposure experiment, 81.8% mortality was recorded. In contrast, no mortality was detected after 10weeks exposure. Moreover, all of the immune system biomarkers studied, except the number of circulating haemocytes, were stimulated by a moderate level of zinc and inhibited by a high level. Our exposure conditions did not induce SOD or MXR mRNA expression in gills and digestive gland. However, an increase of MT mRNA is observed in these tissues. The results indicate that oysters are sensitive to acute zinc toxicity but are only moderately affected by a mild zinc concentration.     

Mn content of reservoir calcite cement: A novel inorganic geotracer of secondary petroleum migration in the tectonically complex Junggar Basin (NW China)

Electronic probe microanalysis(EPMA) results of reservoir calcite cement from fourteen core samples in the Junggar Basin show that Mn-content varies largely between different samples from below the detect limitation to 4.14%,while it displays a generally good correlation with oil-gas shows.This,therefore,likely indicates that concentration of the Mn-content of the calcite cement has a close rela-tion to the intensity of petroleum fluid charging during hydrocarbon secondary migration.In order to assess this hypothesis,oxygen and strontium isotopic measurements on sixteen calcite veins host in source sequences were carried out to investigate the feature of the oil-source petroleum fluid.Analytical results imply that during hydrocarbon generation and migration,deep hot fluid has dissolved volcanic minerals interlined between mudstone source rocks.As Mn is a kind of typical trace element enriched in volcanic rocks,it is reasonable to conclude that the petroleum fluid formed in the source sequences would be Mn-rich.Consequently,calcite cements precipitated from such Mn-rich petroleum fluid would be Mn-rich accordingly.Due to the geologic chromatographic effect during migration along reservoir rocks,the decreasing of the Mn-content of the reservoir calcite cements indicates the migration direction.Then,this novel geotracer was further successfully applied in the study of hydrocarbon migration in the Junggar Basin in combination with organic geochemical analyses during the hydrocarbon migration.The Mn content of the reservoir calcite cement appears promising as a novel inorganic geotracer for the petroleum migration.This paper represents a search for novel indicators of secondary petroleum migration in tectonically complex basins based on fundamentals of the reservoir fluid-rock interactions.     

Hydrocarbon contamination in sediments from urban stormwater runoff

This investigation showed that urban stormwater runoff provides a significant amount of petrogenic material to receiving waters and sediments. A characteristic hydrocarbon ‘fingerprint’ for sediments and particulate matter in the Hillsborough Reservoir, Hillsborough River and upper Hillsborough Bay was provided. Determination of source material for petroleum contamination in stormwater runoff and river sediment indicated that crankcase oil was a primary contributor to sediment hydrocarbon contamination. A comparison of sediment hydrocarbons with hydrocarbons from stormwater runoff showed that the most probable source of crankcase oil-like petrochemicals found in sediment was the stormwater runoff.A comparison of hydrocarbon composition in suspended particulate matter with that of accumulated bottom sediments in the reservoir, river and bay, during a non-storm period and rising tide showed no resuspension and upriver transport of petroleum contaminated bay sediment. No special influence of the bay upon the lower river was observed relative to hydrocarbon tracers, indicating that most contaminated sediment transport was downriver during storm events. Additional studies should be performed over various tidal cycles and storm events incorporating sediment cores, sediment grain size analysis and hydrocarbon characterization at more closely spaced stations near the river mouth to address adequately the question of specific hydrocarbon pollution sources, rate of petroleum influx and persistence of petrochemical contaminants in the sediment.