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The effects of multigenerational Cu exposure on the freshwater gastropod  Isidorella newcombi  were investigated .  Snails were exposed to a range of treatment-specific Cu concentrations in the parental to F 2  generations, and a common Cu concentration in the F 3  generation. In the parental to F 2  generations, some general responses to 3 days Cu exposures included reduced survival and feeding in snails exposed to higher Cu concentrations. This suggested that the snails exposed to the high Cu concentration were experiencing Cu-induced stress that may apply selection pressure. In the F 3  generation, when all treatments were exposed to a common Cu concentration, increased survival was correlated with the pre-exposure Cu concentration history. Snails that had been pre-exposed to Cu also displayed reduced stress at a sub-lethal level, indicated by lower lysosomal destabilisation (LD). Mortality and LD responses in the F 3  generation were not related to Cu tissue concentrations, indicating increased tolerance and reduced stress were not related to changes in Cu bioaccumulation. Total antioxidant capacity increased in the higher Cu concentration pre-exposure treatments which could be associated with lower Cu-induced stress, however, this is not supported by the oxidative damage marker lipid peroxidation, which also increased. While Cu tissue concentrations and oxidative stress markers were assessed to determine underlying reasons for increased tolerance in snails from a population with a multi-generational exposure history to Cu, the results were not conclusive. Despite this, it was demonstrated through increased survival and reduced LD that Cu tolerance can develop over a short evolutionary time scale.

Fitness of Isidorella newcombi Following Multi-generational Cu Exposures: Mortality, Cellular Biomarkers and Life History Responses