The key molecular mechanisms of antagonism induced by combined exposure to erythromycin and roxithromycin in Chlorella pyrenoidosa

Emerging pollutants such as antibiotics have raised great concern in recent years, but the complex coexistence of multiple antibiotics in the environment poses a new challenge in the accurate assessment of the toxicity of antibiotics to aquatic organisms such as microalgae. In this study, the mechanism of action of a combination of erythromycin (ERY) and roxithromycin (ROX) on Chlorella pyrenoidosa was illustrated based on the physiological–biochemical response and transcriptomic analysis. The results revealed an inhibitory effect on the biomass of C. pyrenoidosa at 14 d in all treatment groups, whereas an antagonistic effect was observed in the coexposure groups. The photosystem was the main target despite the existence of multiple compensatory mechanisms, such as expanding the antenna size and initiating alternative electron carriers. The intercept of electrons on the donor side of PSI limited the production of energy, whereas the adjustment of the content and ratio of pigments strengthened microalgal adaptation. Enzymes and genes related to the degradation of exogenous compounds, including cytochrome P450 (P450), glutathione S-transferase (GST) and ABC transporters, mediated the detoxification of antibiotics. The upregulated expression of related genes induced by coexposure increased resistance and explained the antagonistic effects. The shift in energy allocation by increasing the proportion of lipids met the urgent requirements of microalgal physiological activities. This study reemphasizes the modes of interactions between multiple antibiotics and provides new insights into the mechanisms of antagonism induced by combinations of antibiotics.