An Integrated Testing Strategy for Ecotoxicity (ITS‐ECO) Assessment in the Marine Environmental Compartment using Mytilus spp.: A Case Study using Pristine and Coated CuO and TiO₂ Nanomaterials

Abstract

An integrated testing strategy for ecotoxicity assessment (ITS-ECO) was developed to aid in the hazard and fate assessment of engineered nanomaterials (ENMs) deposited in marine environments using the bivalve Mytilus spp. as a test species. The ENMs copper(II) oxide (CuO) and titanium dioxide (TiO 2 ), either in pristine form (core) or with functionalized coatings (polyethylene glycol [PEG], carboxyl [COOH], and ammonia [NH 3 ]) were selected as case study materials based on their production levels and use. High-throughput in vitro testing in Tier 1 of the ITS-ECO revealed CuO ENMs to elicit cytotoxic effects on lysosomes of hemocytes of mussels, with the hazard potential CuO PEG > CuO COOH > CuO NH 3 > CuO core, whereas TiO 2 ENMs were not cytotoxic. Genotoxicity in hemocytes as well as gill cells of mussels following in vivo exposure (48 h) to CuO ENMs was also seen. Longer in vivo exposures in Tier 2 (48 h-21 days) revealed subacute and chronic oxidative effects for both CuO and TiO 2 ENMs, in some cases leading to lipid peroxidation (core TiO 2 ENMs). In Tier 3 bioaccumulation studies, distinct patterns of uptake for Cu (predominantly in gills) and Ti (predominantly in digestive glands) and between the different core and coated ENMs were found. Clear NM-specific and coating-dependent effects on hazard and fate were seen. Overall, using a tiered testing approach, the ITS-ECO was able to differentiate the hazard (acute, subacute, and chronic effects) posed by ENMs of different compositions and coatings and to provide information on fate for environmental risk assessment of these ENMs.