Hepatotoxic mixture exposure

Fish were exposed during 21 days to environmental concentrations of 3 different cyanobacterial hepatotoxic treatments³¹ (CHT), called CHT1-3, as well as to solvent control conditions (Control). The experiment was performed in triplicate tanks for each treatment (comprising 30 fish, 15 males and 15 females per treatment) and the exposure conditions were maintained by the renewal of two-thirds of the total aquaria volume (10 L) containing hepatotoxic mixture every 2 days. Fish were inspected three times daily, and no abnormal behaviour, nor mortality was observed during all the experimentation. At the end of the experiment, fish were anesthetised in 0.1% MS-222, euthanized and then livers were sampled for further analyses using iTRAQ quantitative proteomics.