Fig. 6. Antimicrobial effect of NM102 in vivo in insects and mice.

A P. aeruginosa CIP27853 wt and ∆mfd strains were grown to late exponential phase in LB medium at 37 °C and injected into forth instar silkworm larvae without or with NM102 (2.78 µg/larvae). Boosts of NM102 (6.97 µg/larvae) were administrated at 4 h and 8 h post-infection. At 24 h post-infection larvae were crushed and the content of each tube was serially diluted on LB plates for CFU numeration. Each graph represents the values obtained for n insects with the mean value indicated. Each dot represents one larva (Pa wt n = 10, Pa wt +NM102 n = 10, Pa ∆mfd n = 15, Pa 3/18/25mfd + NM102 n = 15). P values are calculated against the condition wild type-strain without NM102, using Mann-Whitney test (p < 0.0001). For mice experiments, K. pneumoniae ATCC 700603 wt and ∆mfd (B), K. pneumoniae ALE (C), P. aeruginosa CIP27853 (D) were grown to late exponential phase and diluted in PBS. Intranasal (i.n.) bacterial administration was performed through slow instillation of 20 µL of bacterial suspension (1 × 10⁷ CFU). 200 µL of NM102 nanoformulation (NF-NM102 at 1.5 mg/kg or 6 mg/kg) or empty nanoformulation (control) were administrated in mice via the intraperitoneal (i.p.) route (B). Alternatively, 20 µl of a mixture containing bacterial suspension (1×10⁷ CFU) and NM102 nanoformulation (1.5 or 6 mg/kg) or empty nanoformulation was administered via the i.n. route (C, D). MPN (10 mg/kg) was inoculated via the i.p. route. Mice were sacrificed by cervical dislocation after 24 h and the log CFU in the lung was calculated per gram of organ. Each graph represents the bar plot obtained for n mice with mean value indicated: (B) CTR n = 9, NM102 1.5 mg/kg n = 4, NM102 6 mg/kg n = 9, MPN 10 mg/kg n = 6, ∆mfd n = 6, ∆mfd + NM102 n = 10; C CTR n = 8, NM102 1.5 mg/kg n = 8, NM102 6 mg/kg n = 8, MPN 10 mg/kg n = 8; (D) CTR n = 7, NM102 6 mg/kg n = 7, MPN 10 mg/kg n = 8, NM102 1.5 mg/kg+ MPN n = 8, NM102 6 mg/kg + MPN n = 7. P values are calculated using a two-tailed Mann-Whitney test (***p < 0.0005; **p < 0.001; *p < 0.005). The exact P values, minima, maxima, center, bound of box, and whisker are all provided as source data. Source data are provided as a source data file.