K-ras oncogene DNA sequences in pink salmon in streams impacted by the Exxon Valdez oil spill: no evidence of oil-induced heritable mutations.
ABSTRACT It was hypothesized in previous studies that the Exxon Valdez oil spill in Prince William Sound, Alaska, induced heritable mutations and resulted in mortality of pink salmon (Oncorhynchus gorbuscha) embryos. In one of these studies, laboratory exposure of pink salmon embryos to crude oil resulted in apparent mutation-induction in exon 1 and exon 2 of the K-ras oncogene, but no fish from the area impacted by the oil spill were analyzed. We assessed K-ras exon 1 and exon 2 DNA sequences in pink salmon from five streams that were oiled and five streams that were not oiled by the Exxon Valdez oil spill in Prince William Sound, and two streams with natural oil seeps and one stream without seeps on the Alaska Peninsula. Of the 79 fish analyzed for exon 1 and the 89 fish analyzed for exon 2, none had the nucleotide substitutions representing the mutations induced in the laboratory study. Other variable nucleotides occurred in similar proportions in oiled and non-oiled streams and probably represent natural allelic variation. These data do not support the hypothesis that heritable mutations in the K-ras gene were induced by the Exxon Valdez oil spill or oil seeps.
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ABSTRACT: The responses of Dicentrarchus labrax and Liza aurata to aquatic pollution were assessed in a contaminated coastal lagoon, using both traditional and novel biomarkers combined. DNA damage, assessed by comet assay, was higher in both fish species from the contaminated sites, whereas levels of cytochrome P450 1A1 gene expression were not significantly altered. The liver histopathological analysis also revealed significant lesions in fish from contaminated sites. Alterations in ras and xpf genes were analysed and additional pollutant-responsive genes were identified. While no alterations were found in ras gene, a downregulation of xpf gene was observed in D. labrax from a contaminated site. Suppression subtractive hybridization applied to D. labrax collected at a contaminated site, revealed altered expression in genes involved in energy metabolism, immune system activity and antioxidant response. The approach and results reported herein demonstrate the utility of anchoring traditional biomarker responses alongside novel biomarker responses.Environmental Pollution 11/2009; 158(5):1783-90. · 3.73 Impact Factor
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ABSTRACT: The effects of PAHs, considered among the most toxic by the United States Environmental Protection Agency, were tested in vitro and in vivo on two commercial species of the Pertuis-Charentais (Charente-Maritime, France): sea bass, Dicentrarchus labrax, and the Pacific oyster, Crassostrea gigas. This study was carried out as part of the European project EROCIPS with the aim of finding new immunological biomarkers caused by occasional pollution by hydrocarbons. During in vitro experimentation, pollutants and immunological biomarkers were choosen. Thereafter, the in vitro exposures to the soluble fraction of Erika's heavy fuel oil and its fluxant, light cycle oil, began. These exposures enable the validation of the experimental system used, with, in particular, the measurement of bioaccumulated PAHs and metabolites and of choice of the immune biomarkers. The phenoloxidase activity of molluscs and the haemolytic activity of the alternative complement pathway of fish were proposed, for the first time, as suitable biomarkers for the evaluation of pollutant risks in field conditions. Les effets des HAPs, parmi les plus toxiques de la liste de l'Agence de Protection Environnementale Américaine, sont testés in vitro et in vivo sur deux espèces commerciales des Pertuis Charentais (Charente-Maritime, France) : le bar commun, Dicentrarchus labrax, et l'huître creuse, Crassostrea gigas. Cette étude, réalisée dans le cadre du projet européen EROCIPS, recherche de nouveaux descripteurs immunologiques d'une pollution occasionnelle par hydrocarbures. Lors d'expérimentations in vitro, le choix de polluants de type hydrocarbure et de descripteurs d'intérêt de l'immunité non spécifique chez les deux espèces étudiées est réalisé. Puis, des expositions in vivo à la fraction soluble du fioul lourd issus de l'Erika et de son fluxant, le light cycle oil, sont entreprises. Elles ont permis la validation de l'outil expérimental avec notamment la mesure des HAPs bioaccumulés et métabolisés et la détermination d'outils de diagnostic de type immunologique pertinents : l'activité phénoloxydase chez les Mollusques et l'activité hémolytique du complément voie alterne chez les poissons. Ces deux cascades enzymatiques sont proposées pour la première fois dans le cadre d'une évaluation d'une pollution occasionnelle par hydrocarbures pour des conditions réelles de terrain.
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ABSTRACT: The in vitro effects of polycyclic aromatic hydrocarbons (PAHs) on two plasmatic immune parameters, lysozyme concentration and haemolytic alternative complement activity, of the European sea bass, Dicentrarchus labrax, were tested using field (10(-7) and 10(-9) mg mL(-1)) and high concentrations (10(-3) and 10(-5) mg mL(-1)) observed during oil spills. Peripheral blood from 105 fish was collected, centrifuged at 1200 g, for 10 min, at 4 degrees C and three plasma pools, each of 35 fish, were constituted. Two oils (heavy fuel oil and light cycle oil) and 16 pure PAHs, selected on the basis of the American Environmental Protection Agency list (US EPA), were tested in vitro on the two humoral immune parameters. Only three pure PAHs (anthracene, chrysene and dibenz[a,h]anthracene) modulated lysozyme concentration. Acenaphthene, acenaphthylene, anthracene, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, pyrene and light cycle oil modified the haemolytic alternative complement activity after 4h of incubation. This study investigates the direct effects of several PAHs on fish humoral immune functions and describes the haemolytic complement activity of fish as suitable biomarkers of oil pollution.Toxicology in Vitro 03/2009; 23(2):235-41. · 3.21 Impact Factor