Phenotypic and genotypic characterization of Staphylococcus aureus strains isolated from otitis externa: Emergence of CC30-spa t019-SCCmec IV carrying PVL as major genotype

Abstract

The increasing emergence of Staphylococcus aureus as the primary causative agent of otitis externa has been noted; however, detailed information regarding the molecular characteristics of these strains in Iran remains scarce. The current study aims to investigate both genotypic and phenotypic attributes of S. aureus strains implicated in ear infections. In the present work, we analyzed 60 S. aureus strains isolated from cases of otitis externa over a period of 45 months. The resistance patterns were determined using disk diffusion and microbroth dilution methods. All S. aureus isolates were confirmed by the nucA polymerase chain reaction assay, and their biofilm production was assessed by a microtiter plate assay. Molecular characterization of the isolates was performed using the staphylococcal cassette chromosome mec, multilocus sequence typing, and staphylococcus protein A typing methods. Overall, the results indicated that 44 out of 60 S. aureus isolates (73.3%) were methicillin-resistant Staphylococcus aureus. Resistance to mupirocin and vancomycin was observed in 13.3% and 1.7% of the tested isolates, respectively. Furthermore, out of the 60 S. aureus isolates, 56 strains (93.4%) were classified as positive biofilm strains at different levels. Twelve distinct clonal lineages were identified. The vast majority of S. aureus isolates belonged to CC30/ST30-MRSA IV/t019 (41.7%). Among the 31 strong biofilm producers, the majority (64.5%) belonged to CC30/ST30-MRSA IV/t019 clone. Biofilm negative isolates belonged to CC22/ST22 (2 isolates), CC8/ST585 (one isolate), and CC8/ST8 (one isolate). Our result revealed that about three-quarters of PVL-positive strains belonged to CC30/ST30. Our data confirmed the presence of MSSA strains among CC30/ST30 and CC22/ST22 isolates. The mupirocin resistant isolates (n = 8) belonged to CC8/ST585-MRSA III/t713 (37.5%), CC8/ST239-MRSA III/t030 (25%), CC8/ST8-MRSA IV/t008 (12.5%), CC8/ST239-MRSA III/t037 (12.5%), and CC22/ST22-MRSA IV/t790 (12.5%) lineages. The VRSA strain belonged to the CC8/ST8-MRSA IV/t008 lineage, carrying the vanA determinant. iMLSB phenotypes (n = 14) were distributed across different lineages, including CC30/ST30-MRSA IV/t019 (21.5%), CC30/ST30-MSSA/t021 (21.5%), CC22/ST22-MSSA/t005 (14.3%), CC8/ST239-MRSA III/t030 (14.3%), CC22/ST22-MSSA/t1869 (7.1%), CC22/ST22-MRSA IV/t790 (7.1%), CC8/ST239-MRSA III/t037 (7.1%), and CC1/ST772-MRSA IV/t10795 (7.1%). These findings highlight significant genotypic diversity and high biofilm formation among our isolates. The frequent occurrence of the CC/ST30 clone in S. aureus strains isolated from otitis externa reflects the emergence of these lineages as a predominant clone in Iran, posing a significant public health concern.