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In order to study toxicological effects of perfluorononanoic acid (PFNA), perfluorooctane sulfonate (PFOS), and their mixtures (PFNA/PFOS) on Daphnia magna (D. magna), a suite of comprehensive toxicity tests were conducted, including a 48-h acute toxicity test, a 21-day chronic test, a feeding experiment, and a biomarker assay. D. magna were exposed to aqueous solutions of PFNA and PFOS (alone and in combination) at concentrations ranging from 0.008 to 5 mg/L. The survival, growth, and reproduction of D. magna were monitored over a 21-day life cycle. The biomarkers, including acetylcholinesterase (AChE), superoxide dismutase (SOD), and catalase (CAT) activities, were determined after seven days of exposure. PFOS was more toxic than PFNA based on the results of the acute toxicity test. Perfluorinated compounds (PFCs) inhibited both growth and reproduction of D. magna during the testing period. The ingestion rates and the biomarkers, including AChE, SOD, and CAT activities, were significantly inhibited by PFCs in most cases. Moreover, the combined effects related to the growth and reproduction showed the antagonistic effects of PFCs.