Perfluorobutanoic acid (PFBA), an emerging alternative to perfluorooctanoic acid (PFOA), has become increasingly prevalent in aquatic ecosystems, yet its ecotoxicological impacts remain poorly understood. This study investigated the aquatic toxicity of PFBA using the freshwater algae Chlorella vulgaris (C. vulgaris) as a model organism, employing a 96h pre-exposure assay to determine the median effective concentration followed by acute toxicity experiments analyzing multiple endpoints including growth, photosynthetic parameters, oxidative stress markers, and antioxidant enzyme activities. Computer simulation techniques were utilized to illustrate the underlying molecular mechanisms of PFBA toxicity. The results showed that the 96h-EC