The　safety　risks　of　antimicrobial　peptide　(AMP)　resistance　have　never　been　properly　understood.　Herein,　we　comprehensively　investigated　the　evolution　of　resistance　to　AMPs　in　Staphylococcus　aureus　using　piscidin1　(PIS-1)　and　piscidin3　(PIS-3)　as　archetypes,　in　which　they　were　both　combined　with　cell　membranes　via　a　common　structural　motif　but　vary　in　permeabilizing　actions.　The　results　demonstrated　that　the　bacterial　strain　acquired　limited　resistance　to　PIS-3　compared　to　PIS-1.　However,　the　PIS-3-induced　strain　developed　parlous　co-resistance　toward　PIS-1,　ampicillin,　ofloxacin,　rifampicin,　tetracycline,　vancomycin,　and　polymyxin　B.　Based　on　the　results　of　the　chemiluminescence　method,　transcriptome　sequencing　and　proteomic　analysis,　the　generation　of　bacterial　co-resistance　was　affiliated　with　decreased　cell　membrane　permeability,　mainly　involving　the　regulation　of　the　two-component　system,　ATP-binding　cassette　transporters　and　phosphotransferase　system.　These　findings　highlight　concerns　that　AMPs　could　trigger　a　new　crisis　of　drug　resistance　in　food　packaging,　feed　additives,　and　the　agricultural　environment.　©　2022　Elsevier　Ltd