In estuarine ecosystems, bivalves experience large pH fluctuations caused by the anthropogenic elevation of atmospheric CO2 and Cu pollution. This study investigates whether Cu toxicity increases indiscriminately in two bivalve species from different estuarine habitats as a result of elevated Cu bioaccumulation in acidified seawater. This was carried out by evaluating the effects of Cu exposure on two bivalve species (clams and scallops) for 28 d, at a series of gradient pH levels (pH 8.1, 7.8, and 7.6). The results demonstrated an increase in the Cu content in the soft tissues of clams and scallops in acidified seawater. Cu toxicity increased under acidified seawater by affecting the molecular pathways, physiological function, biochemical responses, and health status of clams and scallops. An iTRAQ-based quantitative proteomic analysis showed increased protein turnover, disturbed cytoskeleton and signal transduction pathways, apoptosis, and suppressed energy metabolism pathways in the clams and scallops under joint exposure to ocean acidification and Cu. The integrated biomarker response results suggested that scallops were more sensitive to Cu toxicity and/or ocean acidification than clams. The proteomic results suggested that the increased energy metabolism and suppressed protein turnover rates may contribute to a higher resistivity to ocean acidification in clams than scallops. Overall, this study provides molecular insights into the distinct sensitivities between two bivalve species from different habitats under exposure to ocean acidification and/or Cu. The findings emphasize the aggravating impact of ocean acidification on Cu toxicity in clams and scallops. The results show that ocean acidification and copper pollution may reduce the long-term viability of clams and scallops, and lead to the degradation of estuarine ecosystems.

In order to allow full comparability with other ocean acidification data sets, the R package seacarb (Gattuso et al, 2021) was used to compute a complete and consistent set of carbonate system variables, as described by Nisumaa et al. (2010). In this dataset the original values were archived in addition with the recalculated parameters (see related PI). The date of carbonate chemistry calculation by seacarb is 2022-05-13.The species name used in the paper is Chlamys farreri.