Unraveling the combined toxicity and removal mechanisms of fluoxetine and sertraline co-contaminants by the freshwater microalga Chlorella pyrenoidosa

Selective serotonin reuptake inhibitors (SSRIs), such as fluoxetine (FLX) and sertraline (SER), are among the most widely detected pharmaceuticals in aquatic environments, and they usually occur as mixtures. However, little is known about the combined toxicity of SSRI mixtures to microalgae and the associated removal mechanisms. This study investigated the combined toxicity of FLX and SER to the growth, photosynthetic activity, and antioxidant system of Chlorella pyrenoidosa and their removal mechanisms. The results showed that FLX and SER strongly inhibited microalgal growth with 96 h EC50 values of 493 and 61.1 μg/L, respectively. Additionally, the combined toxicity of FLX and SER towards microalgal growth exhibited an additive effect. After 4 days of short-term exposure, FLX, SER, and their mixtures caused photosynthetic damage and oxidative stress in microalgae, and the mixture's toxicity was stronger than those of individuals. However, the adverse effects on microalgal growth, photosynthetic activity, and antioxidant system were alleviated with increasing exposure time. Meanwhile, C. pyrenoidosa efficiently removed FLX (67.59%–99.08%) and SER (94.92%–99.11%) individually after 11 days of cultivation. Biodegradation (59.25%–86.21%) was the prominent removal mechanism of FLX, while both biodegradation (48.08%–88.17%) and bioaccumulation (4.74%–43.38%) contributed significantly to SER removal. The co-existence of FLX and SER lowered the removal rate and biodegradation amount of both compounds. Besides, SER inhibited C. pyrenoidosa's N-demethylation and O-dealkylation of FLX, while co-existing with FLX inhibited the excretion of the N-deamination product of SER from microalgal cells. Furthermore, the principal component analysis indicated that the removal performance of FLX, SER, and their mixtures correlated strongly to the microalgae's physiological and biochemical states. These results highlighted the significance of co-contamination during ecological risk assessments and microalgae-based bioremediation of SSRIs.