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Genotoxicity of marine sediments in the fish hepatoma cell line PLHC-1 as assessed by the Comet assay
Authors:Maja &Scaron  rut,Luka Traven,Anamaria &Scaron  tambukSonja Kralj,Roko ?ajaVladimir Mi?ovi?,Gö  ran I.V. Klobu?ar
Affiliation:a Department of Zoology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000 Zagreb, Croatia
b Department of Environmental Medicine, Medical Faculty, University of Rijeka, Bra?e Branchetta 20a, 51000 Rijeka, Croatia
c Teaching Institute of Public Health of the Primorsko-Goranska County, Krešimirova 52a, 51000 Rijeka, Croatia
d Laboratory for Molecular Ecotoxicology, Division for Marine and Environmental Research, Ru?er Boškovi? Institute, Bijeni?ka 54, 10000 Zagreb, Croatia
Abstract:The main goal of this study was to test the usefulness of the Comet assay in the PLHC-1 hepatoma fish cell line as a tool for detecting the presence of genotoxic compounds in contaminated marine sediments. The system has been tested using both model chemicals (benzo[a]pyrene (B[a]P) and ethyl methanesulfonate (EMS)) and extracts of sediment samples obtained with solvent dichloromethane/methanol. For all of the analysed sediment extracts as well as for the model chemicals a concentration dependent genotoxic effect was observed. The sediment with the highest observed genotoxic potential was additionally extracted using various solvents in order to test which class of compounds, according to their polarity, is most responsible for the observed genotoxic effect. Non-polar solvents (cyclohexane and dichloromethane) yielded stronger genotoxic effect but the highest level of DNA damage was determined after exposure to sediment extract obtained with the solvent mixture dichloromethane/methanol which extracts a wide range of contaminants. Our results indicate that the PLHC-1 cell line is a suitable in vitro model in sediment genotoxicity assessment and encourage the use of fish cell lines as versatile tools in ecogenotoxicology.
Keywords:Comet assay   PLHC-1 cell line   Marine sediments   DNA damage   Extraction solvent
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