Combined toxic effects of polystyrene microplastics and 3,6-dibromocarbazole on zebrafish (Danio rerio) embryos

Microplastics (MPs) and polyhalogenated carbazoles (PHCZs) are widely detected in the aquatic environment, and their ecological risks have become a research focus. Although there is an extensive co-distribution of MPs and PHCZs, their combined toxicity to aquatic organisms is still unclear. This study investigated the toxic effects of polystyrene microplastics (PS-MPs) and 3,6-dibromocarbazole (3,6-DBCZ) on zebrafish embryos by individual/combined exposure. This study showed that individual or combined exposure of PS-MPs (10 mg/L) and 3,6-DBCZ (0.5 mg/L) could significantly increase the rate of zebrafish embryo deformity, whereas no significant effect was observed on mortality and hatching rate. Furthermore, exposure to 3,6-DBCZ or PS-MPs increased reactive oxygen species (ROS) levels in zebrafish embryos, and the resulting oxidative stress induced apoptosis. Comparably, the levels of oxidative stress and apoptosis in zebrafish embryos were significantly reduced with the combined exposure of 3,6-DBCZ and PS-MPs. These observations suggest that the combined exposure of 3,6-DBCZ and PS-MPs has an antagonistic effect on oxidative stress and apoptosis. Fluorescence PS-MPs tracing and 3,6-DBCZ enrichment analysis showed that, with the protection of chorion, the entry of PS-MPs (5 and 50 μm) into the embryonic stage (55 hpf) of zebrafish was prevented. Moreover, after exposure for 96-144 hpf, PS-MPs served as a carrier to promote the 3,6-DBCZ accumulation and its dioxin-like toxicity in zebrafish larvae through ingestion. Compared with 5-μm PS-MPs, 50-μm PS-MPs promoted higher accumulation and dioxin-like toxicity of 3,6-DBCZ in zebrafish larvae. These findings provide that MPs can be used as an important carrier of PHCZs, influencing their toxicity and bioaccumulation in the organisms.