Correlation between resistance mechanisms in Staphylococcus aureus and cell wall and septum thickening

Abstract

Purpose: The aim of the present study is to examine cell wall and septum thickening of methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), and methicillin- and linezolid-resistant S. aureus (MLRSA) isolates by transmission electron microscopy to correlate the association of resistance mechanisms with major changes in the morphology of membrane or septum.

Materials and methods: MSSA, MRSA, and MLRSA strains obtained from clinical samples of an outbreak that occurred in 2010 at the Intensive Care Unit of our Hospital were thawed and sown at 37°C in blood agar overnight. After that, they were washed, pelleted, and treated with a fixer solution. Pellets were dehydrated and finally embedded in resin. Transmission electron microscopy was used to characterize cell wall and septum thickening in all isolates. The comparison between the measurements obtained for each group was performed by a Kruskal-Wallis test and a post hoc Dunn-Bonferroni's pairwise comparison method.

Results: Differences in cell wall and septum thickness were statistically significant (P<0.001 and P<0.001, respectively) between the three groups. Moreover, significant differences were detected in wall and septum thickness between the MSSA and MRSA strains (P<0.001 and P<0.001, respectively) and between the MSSA and MLRSA strains (P<0.001 and P<0.001, respectively) but not between the MRSA and MLRSA strains (P=0.386 and P=0.117).

Conclusion: In this analysis, we correlate the resistance mediated by alterations in the cell membrane of S. aureus (methicillin-resistant, for example) with a greater thickness of the wall or septum. The resistance added to linezolid did not determine significant changes in the characteristics of the wall or septum with respect to those strains resistant only to methicillin.

Keywords: TEM; VISA; linezolid-resistant S. aureus; protein synthesis inhibitor antibiotics; vancomycin intermediate S. aureus.

Figure 1

Graphical representation of Dunn–Bonferroni’s nonparametric pairwise comparison for post hoc testing after a significant Kruskall–Wallis test of MSSA, MRSA, and MLRSA wall and septum measurements, and TEM images of a representative example of each group. Notes: ^aNumerical values represent the mean rank of each group. Yellow lines represent statistically significant (P<0.05) comparisons between the groups they connect. Black lines represent nonsignificant (P≥0.05) comparisons between the groups they connect. ^bMagnification: 1,50,000×. Scale bar: 200 nm. Abbreviations: MLRSA, methicillin- and linezolid-resistant Staphylococcus aureus; MRSA, methicillin-resistant S. aureus; MSSA, methicillin-sensitive S. aureus; TEM, transmission electron microscopy.