Larval developmental measurements at 96 h post fertilization (hpf) with AFFF exposures. Larval body length (A) and yolk sac area (B) are graphed. (C) Representative images from 0–4.40×10−4% AFFF treatment groups. Bars represent mean±SEM. Asterisk (*) indicate p<0.05 compared with 0% AFFF, one-way ANOVA, Tukey’s HSD post hoc test. n=54–56 larvae per treatment group. Note: AFFF, aqueous film-forming foam; ANOVA, analysis of variance.

The impacts of liver elongation with developmental PFOS and AFFF exposures in 96 h post fertilization (hpf) larvae after developmental PFOS exposures, (A) liver length normalized to larval body length are graphed and (C) representative image are depicted. Scale bars represent 100μM in all images. Following developmental AFFF exposures, (B) liver length normalized to larval body length are graphed and (D) representative image are depicted. Bars represent mean±SEM. Asterisk (*) indicate p<0.05 compared with 0% AFFF, one-way ANOVA, Tukey’s HSD post hoc test. PFOS exposures: n=9 vials. AFFF exposures: n=5–6 vials. Note: AFFF, aqueous film-forming foam; ANOVA, analysis of variance; PFOS, perfluorooctanesulfonic acid.

Exocrine pancreas and endocrine pancreatic beta cell development at 96 h post fertilization (hpf) larvae with AFFF exposure. (A) The area of endocrine pancreatic beta cell islets is graphed. (B) Representative images of normal and fragmented islet morphology in 96 hpf larvae. Separate islet fragments are circled. (C) Pie charts depict the percentage of whole and fragmented beta cell islet morphologies observed. (D) The length of the exocrine pancreas normalized to larval body length is graphed. (E) Individual pancreas lengths are plotted for each treatment group. The 10th (stunted) and 1st (severely stunted) percentile marks are set by control pancreas measurements. (F) Representative images depict a normal exocrine pancreas and severely stunted pancreas in a larva exposed to 4.40×10−4% AFFF. Bars represent mean±SEM. Asterisk (*) indicate p<0.05 compared with 0% AFFF, one-way ANOVA, Tukey’s HSD post hoc test or Fisher’s Exact Test for qualitative assessments. n=6 vials per treatment group. Note: AFFF, aqueous film-forming foam; ANOVA, analysis of variance.