Combined toxicity of polystyrene microplastics and ammonium perfluorooctanoate to Daphnia magna: Mediation of intestinal blockage

Microplastics (MPs) have worldwide accumulated in aquatic environments and coexisted with various water contaminants including perfluorinated compounds (PFCs) that are frequently detected. The adverse effects of individual MPs or PFCs on aquatic organisms have been extensively reported; however, the combined toxicity of MPs and PFCs remains unknown. This study evaluated the combined toxicity of MPs [pristine and aged polystyrene (PS)] and a PFC [ammonium perfluorooctanoate (APFO)] to Daphnia magna under different concentration ratios by three classic methods: toxicity unit, additive index, and mixed toxicity index. The adsorption kinetics of APFO on MPs, aggregation of MPs in exposure medium, MP gut fullness of daphnids, intestinal histology, and lipid peroxidation were analyzed to reveal the mechanism underlying the combined toxicity. Our results showed that the combined toxic modes varied with the concentration ratios of MPs to APFO (antagonism at 4:1 and 1:4, synergism at 3:1, 1:2, and 1:3, and partial addition/antagonism at 2:1 and 1:1 for pristine PS + APFO; antagonism at all ratios except partial addition/antagonism at 3:1 and 1:3 for aged PS + APFO), which could be attributed to the alteration of MP aggregation and thus MP gut fullness in the daphnids. The combined toxicity was further confirmed to occur in the daphnid's gut, which was reflected in physiological and biochemical responses mediated by intestinal blockage. Observable intestinal damages under co-exposures at μg•L levels indicated the risks from future long-term exposure to MPs and PFCs in aquatic environments. This work demonstrates the necessity of assessing combined toxicity with different concentration ratios and provides new insights into the potential risks of MPs in aquatic environments.