Impact of aged and virgin polyethylene microplastics on multi end-points effects of freshwater fish tissues

The buildup of plastic waste in aquatic environments presents serious threats to the environment, wildlife, and ultimately to humans. Specifically, microplastics (MPs) ingestion by aquatic animals leads to adverse physiological and toxicological effects. In addition, discarded MPs undergo aging and degradation processes which affect their morphological properties and chemical composition, enhancing the absorption of environmental pollutants. Under this prism, the present research was conducted to investigate and compare the impact of 'aged' versus pristine low-density polyethylene microplastics (PE-MPs) on various toxicity endpoints as biochemical and molecular parameters in the muscle tissue and liver of the freshwater fish species Perca fluviatilis. In parallel, the morphological, physicochemical, and structural changes occurred in "aged" PE-MPs, (after being exposed to UV radiation for 120 days) were studied, significantly illustrating signs of oxidation and crack propagation at the surface of the studied MPs. Fish were exposed to artificial diet reached with virgin and "aged" PE-MPs, sized 100-180 μm, at concentrations of 1 mg/g of dry food for a period of 15-days. Thereafter, liver and muscle tissues were analyzed in relation to multi oxidative parameters. Compared to the control group, the observed changes in the examined fish included increased activities of antioxidant enzymes, as superoxide dismutase, catalase and glutathione reductase, enhanced concentrations of malondialdehyde, protein carbonylation, HSP70 levels, elevated MAPK phosphorylation, induction of ubiquitin-proteins, as well as heightened levels of Bax/Bcl-2 proteins, caspases and differentiated levels of LC3 II/I, SQSTM1/p62. From the studied biomarkers, apoptosis, ubiquitin and hsp70 levels, showed a more sensitive response against the ingested MPs, followed by autophagy, p38MAPK levels, antioxidant enzymes, MDA and carbonylation levels. The effect of "aged" PE-MPs was more pronounced compared to that of the virgin ones. When evaluating the response of all oxidative stress biomarkers across the studied tissues, the liver demonstrates the highest response for the majority of the biomarkers against both virgin and "aged" PE-MPs. These findings strongly indicate that "aged" MPs activate the antioxidant defence mechanisms and impact the cellular well-being of the examined fish species.