Microplastics (MPs)’ ingestion has been demonstrated in several aquatic organisms. This process may facilitate the hydrophobic waterborne pollutants or chemical additives transfer to biota. In the present study the suitability of a battery of biomarkers on oxidative stress, physiology, tissue function and metabolic profile was investigated for the early detection of adverse effects of 21-day exposure to polystyrene microplastics (PS-MPs, sized 5–12 μm) in the liver and gills of zebrafish Danio rerio and perch, Perca fluviatilis, both of which are freshwater fish species. An optical volume map representation of the zebrafish gill by Raman spectroscopy depicted 5 μm diameter PS-MP dispersed in the gill tissue. Concentrations of PS-MPs close to the EC₅₀ of each fish affected fish physiology in all tissues studied. Increased levels of biomarkers of oxidative damage in exposed fish in relation to controls were observed, as well as activation of apoptosis and autophagy processes. Malondialdehyde (MDA), protein carbonyls and DNA damage responses differed with regard to the sensitivity of each tissue of each fish. In the toxicity cascade gills seemed to be more liable to respond to PS-MPs than liver for the majority of the parameters measured. DNA damage was the most susceptible biomarker exhibiting greater response in the liver of both species. The interaction between MPs and cellular components provoked metabolic alterations in the tissues studied, affecting mainly amino acids, nitrogen and energy metabolism. Toxicity was species and tissue specific, with specific biomarkers responding differently in gills and in liver. The fish species that seemed to be more susceptible to MPs at the conditions studied, was P. fluviatilis compared to D. rerio. The current findings add to a holistic approach for the identification of small sized PS-MPs’ biological effects in fish, thus aiming to provide evidence regarding PS-MPs’ environmental impact on wild fish populations and food safety and adequacy.

The massive accumulation of plastics over the decades in the aquatic environment has led to the dispersion of plastic components in aquatic ecosystems, invading the food webs. Plastics fragmented into microplastics can be bioaccumulated by fishes via different exposure routes, causing several adverse effects. In the present study, the dose-dependent cytotoxicity of 8–10 μm polypropylene microplastics (PP-MPs), at concentrations of 1 mg/g (low dose) and 10 mg/g dry food (high dose), was evaluated in the liver and gill tissues of two fish species, the zebrafish (Danio rerio) and the freshwater perch (Perca fluviatilis). According to our results, the inclusion of PP-MPs in the feed of D. rerio and P. fluviatilis hampered the cellular function of the gills and hepatic cells by lipid peroxidation, DNA damage, protein ubiquitination, apoptosis, autophagy, and changes in metabolite concentration, providing evidence that the toxicity of PP-MPs is dose dependent. With regard to the individual assays tested in the present study, the biggest impact was observed in DNA damage, which exhibited a maximum increase of 18.34-fold in the liver of D. rerio. The sensitivity of the two fish species studied differed, while no clear tissue specificity in both fish species was observed. The metabolome of both tissues was altered in both treatments, while tryptophan and nicotinic acid exhibited the greatest decrease among all metabolites in all treatments in comparison to the control. The battery of biomarkers used in the present study as well as metabolomic changes could be suggested as early-warning signals for the assessment of the aquatic environment quality against MPs. In addition, our results contribute to the elucidation of the mechanism induced by nanomaterials on tissues of aquatic organisms, since comprehending the magnitude of their impact on aquatic ecosystems is of great importance.

Titanium dioxide nanoparticles (TiO2-NPs) are among the most popular manufactured and widely used nanoparticles. They are released into the environment, affecting terrestrial and aquatic ecosystems, with unexpected consequences to organisms and human health. The present study investigates the mediated toxicity imposed to the freshwater fish species, zebrafish (Danio rerio) and the prussian carp (Carassius gibelio), and to the terrestrial land snail Cornu aspersum, after their exposure to sublethal concentrations of TiO2-NPs. Oxidative, proteolytic, genotoxic and apoptotic parameters in fish liver and gills, as well as on snail hemocytes were studied and the swimming performance was estimated in order to (a) estimate and suggest the most susceptible animal, and (b) propose a common battery of biomarkers as the most suitable indicator for biomonitoring studies against TiO2-NPs. Our in vivo experiments demonstrated that NPs induced detrimental effects on animal physiology and swimming behavior, while no general pattern was observed in species and tissues responsiveness. Generally, TiO2-NPs seemed to activate a group of molecules that are common for aquatic as well as terrestrial animals, implying the existence of a conserved mechanism. It seems that after exposure to TiO2-NPs, a common mechanism is activated that involves the stimulation of immune system with the production of ROS, damage of lysosomal membrane, protein carbonylation, lipid peroxidation, DNA damage, following proteolysis by ubiquitin and finally apoptosis. Thus, the simultaneous use of the latter biomarkers could be suggested as a reliable multi parameter approach for biomonitoring of aquatic and terrestrial ecosystems against TiO2-NPs.