Per- and polyfluoroalkyl substances (PFAS) are a class of persistent organic pollutants that pose a growing threat to environmental and human health. Soil acts as a long-term reservoir for PFAS, potentially impacting soil biodiversity and ecosystem function. Earthworms, as keystone species in soil ecosystems, are particularly vulnerable to PFAS exposure. In this study, we investigated the sublethal effects of three short-chain (C4\u2013C6) next-generation perfluoropropylene oxide acids (PFPOAs) on the earthworm Eisenia fetida, using a legacy perfluoroalkyl carboxylic acid (PFCA), perfluorooctanoic acid (PFOA), as a reference. We assessed a suite of biochemical endpoints, including markers for oxidative stress (catalase and superoxide dismutase activity), immunity (phenol oxidase activity), neurotoxicity (acetylcholinesterase activity), and behavioural endpoints (escape test). Results indicate that all tested PFAS, even at sub-micromolar concentrations, elicited significant effects across multiple physiological domains. Interestingly, HFPO-DA demonstrated the most substantial impact across all endpoints tested, indicating broad and significant biochemical and neurotoxic effects. Our findings underscore the potential risks of both legacy and emerging PFAS to soil ecosystems, emphasising the need for further research to understand the long-term consequences of PFAS contamination.