Survival of antimicrobial peptide resistant Staphylococcus aureus in mealworm beetles (Tenebrio molitor) with manipulated AMPexpression

Antimicrobial peptides (AMP) are essential immune effectors of multicellular organisms. Bacteria can evolve resistance to AMPs. Surprisingly, when used to challenge the insect Tenebrio molitor, Staphylococcus aureus resistant to an abundant beetle AMP (tenecin 1) and also pleiotropically to phagocytosis did not increase host mortality or bacterial load compared to infections with wild-type S. aureus. A possible explanation is that antimicrobial resistance is costly due to collaterally increased sensitivity of AMP-resistant strains to other immune effectors. Here, we studied the sensitivity of a group of AMP-resistant S. aureus strains (which are either resistant to tenecin 1 or a combination of both tenecin 1 and tenecin 2) to other immune effectors such as phenoloxidase (PO) and other AMPs (specifically tenecin 1, tenecin 2 and tenecin 4) in vivo. Using RNAi-based knockdown (KD), we investigated S. aureus in vivo survival in insect hosts lacking particular immune effectors. We found that all except one AMP-resistant strain displayed collateral sensitivity toward phenoloxidase. Moreover, some AMP-resistant mutant strains showed sensitivity to one or more components of the beetle AMP defence cocktail. Our findings are consistent with the idea that resistance to AMPs and phagocytosis does not translate into changes in virulence because it is balanced by collaterally increased sensitivity to other host immune effectors. AMP resistance fails to provide a net survival advantage to S. aureus in a host environment that is dominated by AMPs because of the greater sensitivity to other immune effectors.