The role of copper and zinc accumulation in defense against bacterial pathogen in the fujian oyster (Crassostrea angulata).

Cu and Zn are hyper-accumulated in oysters, and the accumulation of these metals increases host resistance to pathogens. However, the role of Cu/Zn in oyster immune defense remains unclear. In this study, Crassostrea angulata with different levels of Cu and Zn were obtained through metal exposure or selective breeding. Both in vivo and in vitro experiments showed that oysters accumulating more Cu/Zn exhibited stronger antibacterial abilities. Vibrio harveyi infection significantly promoted the metal redistribution in oysters: Cu and Zn concentrations decreased in the mantle, but increased in the plasma and hemocytes. This redistribution was accompanied by changes in the expression levels of Cu and Zn transporter genes (CTR1, ATP7A, ZIP1, and ZNT2), suggesting that the Cu/Zn burst observed in the hemocytes was likely due to the transfer of heavy metals from plasma (mediated by the metal importer proteins) or released from intracellular stores. The degree to which Cu/Zn concentration increased in the plasma and hemocytes was more dramatic in oysters with high levels of Cu/Zn accumulation. In vitro, Cu and Zn both inhibited the growth of V. harveyi, while Cu plus H2O2 was lethal to the bacteria. The strength of the growth-inhibition and lethal effects depended on the metal dose. In addition to these effects, increases in Cu concentration increased the activity levels of PO in the oyster plasma and hemocytes in vivo and in vitro. However, SOD activity was not affected by Cu or Zn accumulation. Thus, our results suggested that the Cu/Zn burst in the hemolymph was an important factor in the oyster immune reaction, creating a toxic internal environment for the pathogen, as well as catalyzing inorganic or enzymatic reactions to strengthen bacteriostasis. By determining the extent of Cu/Zn burst in the immune response, Cu/Zn accumulated levels could affect the resistance of oysters to pathogens