Individual and combined toxicity of polystyrene nanoplastics and clothianidin toward <i>Daphnia magna</i>, <i>Lemna minor</i>, <i>Chlamydomonas reinhardtii</i>, and <i>Microcystis aeruginosa</i>

Nanoplastics (NPs) and neonicotinoids are common pollutants in aquatic ecosystems. Although their co-occurrence is expected in multiple environments, studies assessing their combined effects are still limited. This toxicological assessment investigated the potential effects of polystyrene NPs (PSNPs), clothianidin (CLO), and their mixtures on four aquatic species: the freshwater cladoceran Daphnia magna, the duckweed Lemna minor, the green algae Chlamydomonas reinhardtii, and the cyanobacteria Microcystis aeruginosa. Toxicological tests were performed following International Organization for Standardization and Organisation for Economic Co-operation and Development protocols. Acute, chronic (multigenerational) and swimming behavior tests were performed with D. magna, and growth inhibition tests were run with L. minor, C. reinhardtii, and M. aeruginosa. Abbott's model was used to predict the toxicological interactions of the mixtures for each one of the tested species. The D. magna immobility and swimming behavior tests revealed that the combined toxicities of PSNPs and CLO are decreased when the compounds are present as a mixture. Antagonistic interactions were also observed for C. reinhardtii growth, whereas for L. minor and M. aeruginosa, interactions ranged from antagonism to additivity. Chronic multigenerational tests with D. magna revealed that neonates obtained from the exposed parental generation showed a delay in the first brood during the recovery (nonexposure) phase, but this effect disappeared at the next generation, which indicates that microcrustaceans will probably be able to recover on a long-term scale if contamination is stopped. Our results provide new insights into the combined toxicity and ecological risk of NPs and neonicotinoids toward aquatic organisms.