Genotoxic effects of the water-soluble fraction of heavy oil in the brackish/freshwater amphipod Quadrivisio aff. lutzi (Gammaridea) as assessed using the comet assay
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Amphipod crustaceans have been widely used as invertebrate models in ecotoxicology due to their importance in the food chain. However, few studies have evaluated the genotoxic effects of pollutants in this model using the comet assay. The main obstacle to using amphipods in the comet assay is the difficulty in obtaining enough blood cells from a single individual. In this study, we evaluated the genotoxic effects of the water-soluble fraction (WSF) of heavy oil on the brackish/freshwater amphipod Quadrivisio aff. lutzi, which is common in the coastal lagoons of southeastern Brazil, using hemocytes obtained from single amphipods (without pooling) after optimizing hemolymph extraction. The comet assay revealed significantly higher DNA damage levels (2- to 6-fold higher) in treated amphipods compared to untreated ones with a sublethal concentration of 17.6 % of the WSF within 72 h of treatment. Two independent experiments confirmed an “up and down” pattern of DNA damage, measured as the % of DNA contained in the tail of the comets. Elevations in DNA damage levels were observed at the 6 and 48 h time points, while very low levels of DNA damage were observed at the 24 and 72 h time points. Furthermore, the comet assay revealed gender variability in the levels of DNA damage after short-term exposure.
KeywordsEcotoxicology Comet assay Petroleum Invertebrate Amphipoda Gender
The authors are grateful for the financial support that was provided by FINEP/PETROBRAS S/A, Research Fund No. 01.06.1191.00, and to FUNEMAC for three IC Grants. We are thankful to Cristiana Serejo for the taxonomic identification of the amphipods and to Lisia Gestinari for the taxonomic identification of the algae to which the amphipods were associated. We are also particularly grateful to two anonymous reviewers for their valuable suggestions and comments which have improved the manuscript considerably.
Conflicts of interest
The experiments outlined in the present work comply with current Brazilian laws. The authors declare that they have no conflicts of interest.
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