Combined neurotoxicity of aged microplastics and thiamethoxam in the early developmental stages of zebrafish ( Danio rerio )

Microplastics (MPs) pollution, together with its consequential effect on aquatic biota, represent a burgeoning environmental concern that has garnered significant scholarly attention. Thiamethoxam (TMX), a prevalently utilized neonicotinoid insecticide, is renowned for its neurotoxic impact and selective action against targeted pests. The aquatic environment serves as a receptacle for numerous pollutants, such as MPs and neonicotinoid insecticides. However, there is currently a lack of comprehensive understanding regarding the toxic effects of coexposure to aged MPs and neonicotinoid insecticides in aquatic organisms. Therefore, we endeavor to elucidate the deleterious impacts of aged polystyrene (PS) and TMX on zebrafish (Danio rerio) larvae when present at environmentally relevant concentrations, and to reveal the underlying molecular mechanisms driving these effects. Our study showed that exposure to aged PS, TMX, or their combination notably inhibited the heart rate and locomotion of zebrafish larvae, with a pronounced effect observed under combined exposure. Aged PS and TMX were found to diminish the activity of antioxidative enzymes (SOD, CAT, and GST), elevate MDA levels, and disrupt neurotransmitter homeostasis (5-HT, GABA and ACh). Notably, the mixtures exhibited synergistic effects. Moreover, gene expression related to oxidative stress (e.g., gstr1, gpx1a, sod1, cat1, p38a, ho-1, and nrf2b) and neurotransmission (e.g., ache, ChAT, gat1, gabra1, 5ht1b, and 5ht1aa) was significantly altered upon co-exposure to aged PS and TMX in larval zebrafish. In summary, our findings support the harmful effects of aged MPs and the neonicotinoid insecticides they carry on aquatic organisms. Results from this study enhance our understanding of the biological risks of MPs and insecticides, as well as help fill existing knowledge gaps on neonicotinoid insecticides and MPs coexistence toxicity in aquatic environment.