ABSTRACT
Significant progress has been made in developing fluorine-free firefighting foams (F3) as alternatives to perfluoroalkyl substances (PFAS)-containing aqueous film-forming foams (AFFF) to help eliminate the health and environmental concerns linked to PFAS exposure. However, developing viable F3 options hinges on a thorough assessment of potential risks alongside the technical performance evaluations. This study showcases the capability of a zebrafish-based platform to discern the developmental and behavioral toxicities associated with exposure to one AFFF and two F3 formulations. To facilitate direct exposure to the chemicals, embryos were enzymatically dechorionated and then exposed to the diluted formulations (6-120 hours post fertilization (hpf)) at concentrations folding from 0.1% of the manufacturer-recommended working concentrations. The exposure regimen also included daily automated media changes (50%) and mortality assessments (24 and 120 hpf). At 120 hpf, a comprehensive assessment encompassing overall development, prevalence of morphological defects, and behavioral responses to acute stressors (visual, acoustic, and peripheral irritant) was conducted. Exposure to both F3s significantly increased larval mortalities to percentages exceeding 90%, whereas AFFF exposures did not cause any significant effect. Overall development, marked by total larval length, was significantly impacted following exposures to all foams. Behavioral responses to acute stressors were also significantly altered following exposures to both F3s, whereas the AFFF did not alter behavior at the concentrations tested. Our findings demonstrate toxicities associated with tested F3 formulations that encompass several endpoints and highlight the utility of the proposed platform in evaluating the developmental toxicities of current and future foam formulations.