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Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
Published on: September 8, 2021
Effects of the antimicrobial peptide L12 against multidrug‑resistant Staphylococcus aureus
Fu Xiong1, Xiaotian Dai1, Yu-Xue Li2
1Department of Pulmonology, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing 400038, P.R. China.
Abstract:
Methicillin‑resistant Staphylococcus aureus (S. aureus; MRSA) is one of the most common bacterial pathogens and MRSA infections are characterized by high mortality rates. Antimicrobial peptides are considered one of the most promising drugs for the treatment of resistant strains of S. aureus. The present study aimed to examine the antimicrobial activity of L12 against numerous bacterial species using the broth microdilution method. Furthermore, the synergistic effect of L12 combined with various antibacterial drugs was tested, and its antibacterial mechanism was investigated by a checkerboard assay. The alterations in bacterial morphology were detected by electron microscopy, and biofilm formation and removal were tested by crystal violet staining. The present results suggested that L12 affected the growth of gram‑positive strains, particularly S. aureus. Electron microscopy analysis suggested that L12 may target the cell membrane, and L12 increased the antibacterial activity of vancomycin and levofloxacin, exerting a synergistic effect. However, the minimal inhibitory concentrations (MICs) of L12 were not correlated with antibiotic resistance, the strains resistant to more antibiotics were not more resistant to L12. A sub‑MIC of L12 was able to inhibit biofilm formation in a dose‑dependent manner; however, concentrations of L12 ≤10 times the MIC were not sufficient to degrade previously formed biofilm. Collectively, the present study suggested that L12 may represent a novel potential therapeutic molecule for the treatment of S. aureus infections.
Insights
The antimicrobial peptide L12 shows potent activity against Gram-positive bacteria, particularly Methicillin-resistant Staphylococcus aureus (MRSA). L12 enhances antibiotic effectiveness and inhibits MRSA biofilm formation, offering a potential new treatment for MRSA infections.
Area of Science:
- Microbiology
- Pharmacology
- Biochemistry
Background:
- Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant threat due to high mortality rates.
- Antimicrobial peptides are a promising avenue for combating resistant bacterial strains.
Purpose of the Study:
- To evaluate the antimicrobial activity of the peptide L12 against various bacteria.
- To investigate the synergistic effects of L12 with existing antibiotics.
- To elucidate the mechanism of action and anti-biofilm properties of L12.
Main Methods:
- Broth microdilution method for antimicrobial activity testing.
- Checkerboard assay for synergy testing.
- Electron microscopy for morphological analysis.
- Crystal violet staining for biofilm assessment.
Main Results:
- L12 demonstrated antimicrobial activity, especially against Gram-positive bacteria like S. aureus.
- L12 exhibited synergistic effects, enhancing the activity of vancomycin and levofloxacin.
- Electron microscopy suggested L12 targets the bacterial cell membrane.
- L12 inhibited biofilm formation in a dose-dependent manner but did not degrade pre-formed biofilms.
Conclusions:
- L12 shows potential as a therapeutic agent against S. aureus infections.
- L12's mechanism involves cell membrane disruption and biofilm inhibition.
- L12's efficacy is not directly correlated with existing antibiotic resistance levels.
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