Azithromycin induces dual effects on microalgae: Roles of photosynthetic damage and oxidative stress

Antibiotics are frequently detected in aquatic ecosystems, posing a potential threat to the freshwater environment. However, the response mechanism of freshwater microalgae to antibiotics remains inadequately understood. Here, the impacts of azithromycin (a broadly used antibiotic) on microalgae Chlorella pyrenoidosa were systematically studied. The results revealed that high concentrations (5-100 μg/L) of azithromycin inhibited algal growth, with a 96-h half maximal effective concentration of 41.6 μg/L. Azithromycin could weaken the photosynthetic activities of algae by promoting heat dissipation, inhibiting the absorption and trapping of light energy, impairing the reaction centre, and blocking electron transfer beyond Q_A. The blockage of the electron transport chain in the photosynthetic process further induced the generation of reactive oxygen species (ROS). The increases in the activities of superoxide dismutase, peroxidase and glutathione played important roles in antioxidant systems but were still not enough to scavenge the excessive ROS, thus resulting in the oxidative damage indicated by the elevated malondialdehyde level. Furthermore, azithromycin reduced the energy reserves (protein, carbohydrate and lipid) and impaired the cellular structure. In contrast, a hormesis effect on algal growth was found when exposed to low concentrations (0.5 and 1 μg/L) of azithromycin. Low concentrations of azithromycin could induce the activities of the PSII reaction centre by upregulating the mRNA expression of psbA. Additionally, increased chlorophyll b and carotenoids could improve the absorption of light energy and decrease oxidative damage, which further contributed to the increase in energy reserves (protein, carbohydrate and lipid). The risk quotients of azithromycin calculated in this study were higher than 1, suggesting that azithromycin could pose considerable ecological risks in real environments. The present work confirmed that azithromycin induced dual effects on microalgae, which provided new insight for understanding the ecological risk of antibiotics.