Fig. 1

E-liquid effects on development, fecundity, and vitality in C. elegans. Synchronized populations of N2-Bristol strain nematode embryos were placed onto OP50 bacterial seeded NGM agar plates supplemented with PG (PG) or e-liquids containing grape (G), menthol (M) or V2 Red (R) additives, ± nicotine, as indicated. The worms were allowed to mature on these plates and physiologic effects were determined. a The life cycle of C. elegans consists of four larval stages prior to reproductive adulthood, and at 20 °C worms take ~3 days to mature. b Body length was assessed every day, with the ~60 h data shown. c Brood size was determined over the entire fertile period, starting from day 1 of adulthood (~60 h) through day 5. d Thrashing rates were measured using post-hoc analysis of video acquired following movement of the worms from plates to liquid M9 media, first at ~60 h and again after 7 days. Each measurement consists of at least three independent experimental replicates. The number of individual worms analyzed per replicate was from 10–20 for body size and 5 for brood size and thrashing. Two-way ANOVA with Bonferroni post-hoc analysis was used for statistical analysis. A hashtag (#) represents significant differences (p < 0.05) between the negative control (C) and treatments lacking nicotine, and an asterisk (*) is used to represent significant differences (p < 0.05) between the nicotine control (C+) and treatments containing nicotine. Though not denoted, the statistical analysis also indicated a main effect for nicotine, but no interaction between nicotine and the solvent groups. All 7 day values for thrashing differed significantly from the day 3 values but not among themselves. ND is not determined