2.11. Galleria Mellonella Larvae Infection Model

The virulence of PAO1 mutants resistant to PA5oct phage was tested in vivo in G. mellonella infection model, previously described by Cullen et al. [34]. The wax moth larvae were sorted by size and weight and acclimated for one week at 15 °C. The larvae were infected by the injection into the hindmost proleg, of 10 µL of bacterial suspension (10³ cfu/mL) giving 10 cells per larvae. Next, the larvae were incubated at 37 °C for 72 h, and their viability was checked after 8, 18, 24, 48 and 72 hours post-injection. The two types of control were made: the negative (larvae injected with 10 µL of sterile PBS buffer) and positive (larvae infected with a wild PAO1 strain). The graphical presentation of the survival curves was prepared using GraphPad Prism 6.0 (GraphPad Software Inc., La Jolla, CA, USA). The log-rank Mantel-Cox test was used for statistical analysis (P-values < 0.05 were regarded as significant).