Genotoxicity assessment in the amphipod Gammarus fossarum by use of the alkaline Comet assay

Abstract

Many xenobiotics and newly developed substances released in the aquatic environment have been found genotoxic for living organisms. There is interest in developing biomarkers of genotoxicity in different phyla and the need to increase our understanding of the impact of genotoxic insult on invertebrates, particularly on crustaceans. Freshwater invertebrates and particularly amphipods are highly relevant species ecologically. However, genotoxic responses of such species are rarely studied, whereas understanding these responses is becoming an urgent concern. The aim of this study was to develop and optimize the Comet assay in the freshwater invertebrate Gammarus fossarum by use of different cell-types: haemocytes, oocytes and spermatozoa. In a first step, the Comet assay was performed on these three cell types after exposure to the model genotoxicant methyl methanesulfonate (MMS) in vitro and in vivo. Results showed a clear dose-response relationship for all tissues, a low variability and a high sensitivity of the response, demonstrating the effectiveness of the Comet assay to detect genotoxic insult in amphipods. In a second step, to explore the potential of this technique for use in ecotoxicological studies with amphipods, these organisms were exposed to five known or suspected genotoxic compounds. The results demonstrated the possibility to use the freshwater amphipod G. fossarum in environmental genotoxicity studies with the Comet assay.