Multigenerational effects of virgin and sampled plastics on the benthic macroinvertebrate Chironomus riparius

Although sediments are important reservoirs of plastics, most of the ecotoxicological studies on these contaminants are focused on the organisms living in the water column, while only a smaller number of evidence concerns the plastic impact on benthic species. Therefore, this study compared the multigenerational effects on the sediment-dwelling midge Chironomus riparius exposed to both virgin polystyrene microbeads (22,400-224,000 plastics/kg sediments dry weight), and plastic mixtures (40-420 plastics/kg dry weight) collected from four of the main tributaries of Po River (Ticino, Adda, Oglio and Mincio Rivers, Northern Italy) to evaluate the role played by other characteristics related to these physical contaminants in determining their toxicity as opposed to concentration alone. The modified Chironomid Life-Cycle Toxicity Test (OECD 233) was used to evaluate the multigenerational effects on the Emergence and Development Rates, Fecundity and Fertility. In addition, a biomarkers' suite of cellular stress, neurotoxicity, and energetic metabolism was applied in the 2^nd generation (2^nd/3^rd instar of larvae) to investigate the potential mechanisms associated to the apical effects. Our results showed no significant (p > 0.05) multigenerational effect for any of the endpoints tested for the virgin plastics' exposures. Coherently, no significant effects on biomarkers were measured. Concerning the sampled plastics, the particles collected in Adda River instead induced a significant decrease (p < 0.05) of the Emergence Rate in the 2^nd generation, suggesting that this parameter was the most susceptible among those measured. These results highlight that the different plethora of polymers, sizes and shapes of plastics sampled in natural ecosystems, compared to homogeneous characteristics of virgin polystyrene microbeads, appears to have considerable importance over concentration alone in determining the toxicity of these emerging contaminants.