Clinical observations of BALB/c mice during the vaccination protocol. Female BALB/c mice had corporal temperatures recorded immediately before and 24 h after each (A) intramuscular (IM)/intranasal (IN) vaccination or (B) IM vaccination event. Mice were also weighed once a week starting on day 0 (first vaccination) until day 63 of the vaccination protocol for the (C) IM/IN vaccinated and the (D) IM vaccinated mice. Data for each figure are represented as ± SEM, (A) and (C) n = 15, (B,D) n = 5 for each group. Statistical analyses for (A) were determined by an unpaired two-tailed T-test, % p = 0.0255 and ****, ####, $$$$ p < 0.0001 as compared to the 24 h prior measurement for each dose, respectively. Statistical analyses for (B) was determined by an unpaired two-tailed T-test for % p = 0.0285, ## p = 0.012, *** p = 0.0007 as compared to the 24 h prior measurement for each dose/group respectively and a Mann–Whitney U-test for $ p = 0.0442 as compared to the 24 h prior measurement.