Effects of copper on hemocyte parameters in the estuarine oyster Crassostrea rivularis under low pH conditions

With the development of industry and agriculture, the metal pollutants (e.g., Cu) are inevitably released into the aquatic environment. In addition, ocean acidification (OA) as a major environmental stress is affecting marine organisms. In this study, we investigated the hemocyte responses of the estuarine oyster Crassostrea rivularis exposed to six combinations of two pH levels (8.1 and 7.7) and three Cu concentrations (0, 10 and 50 μg/l) using flow cytometry in vitro and in vivo. In both experiments, Cu and low pH jointly affected the hemocyte parameters of oyster. High Cu exposure resulted in decreased total hemocyte count (THC), esterase activity (EA) and lysosomal content (LC) and increased hemocyte mortality (HM), phagocytosis activity (PA) and reactive oxygen species (ROS) production, especially under low pH conditions. The immune suppression of metal-exposure was more significant than low pH exposure with a 28-d experimental period in oysters. A slight recovery of the immune parameters was observed in THC, HM, PA, ROS and LC. During the depuration period, the modulatory effects of pH were still obvious. In addition, carry-over effects of high Cu and low pH were still observed. Overall, our results showed that copper and low pH weaken immune functions of hemocyte in oysters, with synergistic effects. This work provides new evidence of sublethal negative effects of metals on marine animals under global change scenarios, and copper likely leads to reduced fitness of oysters under low pH conditions.