Immunotoxic effects of exposure to the antifouling copper(I) biocide on target and nontarget bivalve species: a comparative in vitro study between Mytilus galloprovincialis and Ruditapes philippinarum

Front Physiol. 2023 Aug 15:14:1230943. doi: 10.3389/fphys.2023.1230943. eCollection 2023.


Edible bivalves constitute an important bioresource from an economic point of view, and studies on their immune responses to environmental pollutants are crucial for both the preservation of biodiversity and economic reasons. The worldwide diffusion of copper(I)-based antifouling paints has increased copper leaching into coastal environments and its potential impact on both target and nontarget organisms. In this study, immunotoxicity assays were carried out with short-term (60 min) cultures of hemocytes from the bivalves Mytilus galloprovincialis-a mussel dominant in the macrofouling community-and Ruditapes philippinarum-a clam dominant in the soft-sediment community-exposed to CuCl to compare the toxic effects on their immune responses. The LC50 values were similar, 40 μM (3.94 mg L-1) for the mussel and 44 μM (4.33 mg L-1) for the clam. In both species, apoptosis occurred after exposure to 1 µM (98.9 μg L-1) CuCl, the concentration able to significantly increase the intracellular Ca2+ content. Biomarkers of cell morphology and motility revealed microfilament disruption, a significant decrease in yeast phagocytosis and lysosome hydrolase (β-glucuronidase) inhibition beginning from 0.5 µM (49.5 μg L-1) CuCl in both the mussel and clam. The same concentration of CuCl affected biomarkers of oxidative stress, as a significant decrease in reduced glutathione content in the cytoplasm and inhibition of mitochondrial cytochrome-c oxidase (COX) were detected in both species. Comparison of the biomarkers showed that clam is more sensitive than the mussel regarding alterations to the lysosomal membrane and reactive oxygen species (ROS) production, which supports the potential harmful effects of antifouling biocides on the survival of nontarget pivotal species in the coastal community.

Keywords: Mytilus galloprovincialis; Ruditapes philippinarum; antifouling paints; bivalves; copper(I) chloride; hemocytes; immunotoxicity; mollusks.

Grants and funding

This research was supported by grants from the Italian MIUR (DOR 2021) to FC and by a funding agreement between P. Zarantonello’s “Resimix s. r. l. ” (Brendola, Vicenza, Italy, (accessed on 20 May 2023)) and the University of Padova, Italy for a Ph.D. fellowship in Biosciences to RV (Rep. #1488, Prot. #186788, 6 May 2019).