The emergence of novel macrolide resistance island in Macrococcus caseolyticus and Staphylococcus aureus of food origin

Abstract

Food-derived Staphylococcaceae species with severe antimicrobial resistance, especially Staphylococcus aureus, is a major threat to public health. Macrococcus caseolyticus (M. caseolyticus) is a member of the Staphylococcaceae family which plays a vital role in fermented products and disease causation in animals. In our previous study, several Staphylococcus aureus antibiotic-resistant island msr (SaRI_msr) were found in multidrug-resistant S. aureus. In this study, novel SaRI_msr, SaRI_msr-III emerged from S. aureus. Another novel SaRI_msr-like further emerged in M. caseolyticus from food. These isolates' prevalence and genetic environment were investigated and characterized to understand the distribution and transmission of these novel SaRI_msr strains. All SaRI_msr-positive S. aureus isolates exhibited a multidrug resistance (MDR) phenotype, within which a series of antimicrobial resistance genes (ARGs) and virulence factor genes (VFs) were identified. In addition, three SaRI_msr types, SaRI_msr-I (15.1 kb), SaRI_msr-II (16-17 kb), and SaRI_msr-III (18 kb) carrying mef(D)-msr(F), were identified in these isolates' chromosomes. SaRI_msr-(I-III) contains a site-specific integrase gene int and operon mef(D)-msr(F). SaRI_msr-III has an additional orf3-orf4-IS30 arrangement downstream of mef(D) and msr(F). Moreover, the SaRI_msr-like and macrolide-resistant transposon Tn6776 forming a novel mosaic structure coexisted in one M. caseolyticus isolate. Within this mosaic structure, the macrolide-resistant genes mef(D)-msr(F) were absent in SaRI_msr-like, whereas an operon, mef(F)-msr(G), was identified in Tn6776. The SaRI_msr-(I-III) and SaRI_msr-like structure were inserted into the rpsI gene encoding the 30S ribosomal protein S9 in the chromosome. Excision and cyclisation of SaRI_msr-III, SaRI_msr-like, operon mef(D)-msr(F), and orf3-orf4-IS30 arrangements were confirmed using two-step PCR. This study is the first to report MDR S. aureus harbouring novel SaRI_msr-III and M. caseolyticus containing novel mosaic structures isolated from retail foods. Similar SaRI_msr-type resistant islands' occurrence and propagation in Staphylococcaceae species require continuous monitoring and investigation.

Keywords: Antibiotic resistant islands; Macrolides; SaRI(msr)-like; Tn6776; mef(D)-msr(F).