Synergistic toxicity of microcystin-LR and Cu to zebrafish (Danio rerio)

Toxic cyanobacterial blooms often coincide with metal pollution in a freshwater environment because of surface run-off enriched with nutrients and metals. However, the joint toxic effects of cyanobacterial toxins and metals on aquatic animals remain unknown. In this study, single and joint toxic effects and mechanisms of microcystin-LR (MCLR) and copper (Cu) were investigated in the early development of zebrafish (Danio rerio). The LC50_72-h values were 2.79 mg/L for MCLR and 3.23 mg/L for Cu. The sublethal concentrations of MCLR (≤600 μg/L) did not affect the normal development of zebrafish but increased its hatchability. Strong synergistic toxic effects were observed after co-exposure to MCLR and Cu at environmental concentrations (≤60 μg/L). The synergistic toxic effects of these two compounds could be attributed to the increased bioaccumulation of MCLR and Cu, which was mediated by MCLR transporters (e.g., oatp1d1 and oatp2b1) and Cu transporters (e.g., ctr1 and atp7a), in zebrafish. Such bioaccumulation caused oxidative stress, as suggested by the disrupted gene expression of anti-oxidative enzymes (e.g., Cu/Zn-SOD, Mn-SOD, and CAT). Our results revealed for the first time the synergistic toxic effects and potential toxic mechanism of MCLR-Cu in aquatic animals. These synergistic effects should be considered when assessing the ecological risk of toxic cyanobacterial blooms.