Objectives: The purpose of this study was to investigate the effect of Boesenbergia rotunda (L.) Mansf. extract (BRE) and peptidoglycan inhibitor antibiotics, alone and in combination, against β-lactam-resistant staphylococci.
Methods: Antibacterial and synergistic activities of BRE alone and in combination with ampicillin (AMP), cloxacillin (CLX), cefazolin (CZO) or vancomycin (VAN) were evaluated against two β-lactam-resistant Staphylococcus aureus (BRSA) isolates and one β-lactam-resistant Staphylococcus epidermidis (BRSE) isolate. The activities were confirmed by killing curve assays. The preliminary antimicrobial action was elucidated by transmission electron microscopy (TEM) and cytoplasmic membrane (CM) permeability assay.
Results: All tested staphylococci were inhibited by BRE at a minimum inhibitory concentration (MIC) of 16μg/mL. Two BRSA strains showed high resistance to CLX, AMP and CZO, whilst BRSE was resistant to CLX and AMP. All tested isolates remained susceptible to VAN. Chequerboard assay demonstrated a fractional inhibitory concentration index (FICI) of 0.50 for the BRE+CLX combination against the BRSA strains. Killing curve determinations confirmed the antibacterial and synergistic activities. TEM revealed collapse of the CM in BRE-treated cells and damage both of the CM and peptidoglycan (PG) in BRE+CLX-treated cells. The CM permeability assay showed that either BRE or nisin alone as well as BRE+CLX significantly induced leakage of OD260nm-absorbing materials.
Conclusions: BRE potentiated the activity of β-lactams, particularly CLX, against β-lactam-resistant staphylococci by damaging the CM and PG layer, leading to leakage of intracellular material. Combination of BRE and β-lactams provides a potential way forward in developing novel antistaphylococcal agents.
Keywords: Boesenbergia rotunda (L.) Mansf. extract; Cytoplasmic membrane; Staphylococci; β-Lactams.
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