Effect of Bicomponent ZnO-ZnFe₂O₄ Nanoparticles on Mediterranean Mussel (Mytilus galloprovincialis) Hemocytes under in vitro Conditions

The present work investigates the toxic effect of bicomponent ZnO-ZnFe₂O₄ nanoparticles, which are the main active component of the domestic antifouling coating, on marker indicators of Mediterranean mussel (Mytilus galloprovincialis) hemolymph cells (hemocytes) under in vitro experimental conditions. The following indicators were evaluated: mortality, cellular composition and production of reactive oxygen species. In the experiment, hemocytes were incubated for 1 and 2 hours in 1 mL of sterile seawater containing nanoparticles of different concentrations: 0.03, 0.3 and 3 mg/mL. The data were analyzed using the flowing cytometry. It was shown that ZnО-ZnFe₂O₄ nanoparticles had an effect on the cellular composition of the hemolymph: the proportion of agranulocytes decreased and hour exposure to 0.03 mg/mL nanoparticles reduced the level of production of reactive oxygen species by 2.5 times compared to the control (p ≤ 0.05). Incubation of hemocytes with a maximum concentration of nanoparticles (3 mg/mL) led to cell death within 1 hour after exposure. No acute toxic effects on hemocytes with the use of 0.03 mg/mL and 0.3 mg/mL of zinc oxide and zinc ferrite nanoparticles were observed.