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Methicillin-resistant Staphylococcus aureus (MRSA) biofilm infections are difficult to treat due to the high antimicrobial resistance of biofilm. Therefore, new treatments are needed for more effective bacteria clearance. This study was to investigate whether low frequency ultrasound (LFU) can enhance the activity of antimicrobial agents against MRSA biofilm infection. Broth microdilution method was used to determine the minimum inhibitory concentration (MIC) of vancomycin (VAN), linezolid (LIN), and levofloxacin (LEV) against three clinical isolated strains, including one methicillin-susceptible Staphylococcus aureus (MSSA) strain and two MRSA strains. Effects of various influencing factors, such as antimicrobial agents, drug concentrations, ultrasonic intensity, and single (S-LFU, 5 or 15 min) or multiple ultrasound (M-LFU, 5 min every 8 h), on the inhibition of biofilms were investigated. The bactericidal effects of S-LFU or M-LFU on MRSA or MSSA biofilms were determined by colony counts. Right after ultrasound, synergistic effects were observed in groups of S-LFU combined with three antimicrobial agents against MSSA biofilm, but for MRSA biofilm, only S-LFU plus VAN had synergistic effect. At the time point of 24 h, M-LFU plus VAN treatment had synergistic bactericidal effect against MRSA and MSSA biofilms, and the synergy showed that VAN is concentration-dependent, but no synergistic effects were observed in all S-LFU combination groups. In conclusion, combination of M-LFU and antimicrobial agents had a better synergistic effect than S-LFU against MRSA or MSSA biofilm. LFU may be useful in treating biofilm infection in the future.

Multiple Low Frequency Ultrasound Enhances Bactericidal Activity of Vancomycin against Methicillin-Resistant Staphylococcus aureus Biofilms