Figure 5

Assessment of BIMFJ1 phenotypic costs. A Bacterial viability in pig serum and adhesion to IPEC-1 cultured cells. Bars indicate the concentration (CFU mL⁻¹) of each strain (BIM 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.10 and EC43 WT) before and after incubation with pig serum. Dots represent the concentration (CFU cm⁻²) of each strain (BIM 1.1, 1.4, 1.6, 1.9, 1.10 and WT EC43 adhered to IPEC-1, 2 h after infection (MOI = 100). Errors bars represent standard deviation for an average of three repeated experiments. Statistical significance of differences between BIMs and the WT strain was determined using a one-way ANOVA test (* p-value < 0.05** p-value < 0.01; *** p-value < 0.001; **** p-value < 0.0001); B Health index in G. mellonella model. Larvae health index scores in wax moth larvae, 24 h, 48 h and 72 h post-infection with BIM 1.1, 1.6, 1.10 and EC43 and PBS (control). Statistical significances of differences between mutants, wild-type and control groups were calculated using a two-way ANOVA (* p-value < 0.05; ** p-value < 0.01; *** p-value < 0.001; **** p-value < 0.0001); C Survival rates of challenged G. mellonella. Larval survival rates, 24 h, 48 h and 72 h post-infection with BIM 1.1, 1.6, 1.10 and EC43 and PBS (control). All experiments were performed in triplicate and in a minimum of three independent assays. Statistical significance was calculated using the log-rank Mantel-Cox statistical test (**** p-value < 0.0001).