Perfluorooctane sulfonamide (PFOSA), a typical perfluorooctane sulfonate precursor (PreFOS), has been detected in the aquatic environment globally. However, the effects of PFOSA at levels measured in the environment have not been well characterized in aquatic organisms. In this study, we evaluated the transcriptional, biochemical, histopathological, and morphological effects of PFOSA to characterize the underlying mechanisms of toxicity by using a universal model in aquatic ecotoxicology, zebrafish (Danio rerio). Transcriptional changes in PFOSA-exposed zebrafish predicted hepatic fibrosis and associated immune function. Subsequent, sublethal impacts were observed, which included significant alterations in liver-specific protein levels, increased immune cell numbers, and liver pathological structural damage. In addition, we compared the effects caused by PFOSA and perfluorooctane sulfonate (PFOS) at the same exposure concentration and found a greater hepatotoxic effect of PFOSA relative to PFOS, indicating that the adverse impacts of PFOSA may be more severe. This was the first study to comparatively explore the hepatotoxic response of PFOSA and PFOS in aquatic organisms, which can be used for ecological risk assessments of PreFOS compounds.