Due to the increased emergence of drug-resistant bacteria, the declining efficiency of traditional antimicrobials has generated severe concerns in recent years. Subsequently, more interest in other antimicrobial agents from natural resources draws more attention as an alternative to conventional medications. This study investigated the bactericidal mechanism of monoterpene 1,8-cineol (eucalyptol), a major compound of various essential oils, against methicillin-resistant Staphylococcus aureus (MRSA). The antibacterial activity of 1,8-cineol was assessed by an MTT assay against clinical and reference MRSA strains. A cell membrane integrity test, followed by zeta potential (ZP) measurements, was performed to evaluate the disruption of the bacterial membrane integrity. Additionally, the cytotoxic effect of this molecule on MRSA bacteria was investigated by monitoring reactive oxygen species (ROS) generation, lipid peroxidation (MDA), and antioxidant enzyme activities (CAT and SOD). Regarding the anti-staphylococcal effect, the obtained results revealed the antibacterial efficacy of 1,8-cineol wherein the minimum inhibitory concentrations were equal to 7.23 mg/mL. Furthermore, it enhanced membrane permeability, with a 5.36-fold increase in nucleic acid and protein leakage as compared with untreated strains, along with the alteration of surface charge (ZP) in MRSA cells. The tested compound caused an increase in ROS generation reaching 17,462 FU and MDA production, reaching 9.56 μM/mg protein, in treated bacterial cells, along with a decrease in oxidative stress enzymes activities. Our findings suggest that 1,8-cineol has the ability to damage the membrane integrity and induce ROS-mediated oxidative stress in MRSA cells, leading to its antagonistic effect against this pathogen and consequently aiding in the reversal of antibiotic resistance.

Background: Lamiaceae family is one of the most diverse and common plant families in terms of ethnomedicine due to their potential therapeutic effects. The aim of this study is to investigate the correlation between the chemical composition and the antibacterial effect of five essential oils from this family against five reference bacterial strains responsible of nosocomial diseases and foodborne illnesses. Methods: The commercial essential oils of Tunisian Rosmarinus officinalis, Thymus capitatus, Origanum majorana and Salvia officinalis were analyzed by GC/FID and GC-MS. Essential oils were evaluated for their antibacterial activities by disc diffusion and microbroth dilution methods against five reference bacterial strains: Pseudomonas aeruginosa, Escherichia coli, Salmonella enterica, Bacillus subtilis and Staphylococcus aureus. The inhibition zone diameter values and the twenty major compounds of the selected essential oils were subjected to PCA and HCA analysis. Results: Analysis by GC/FID and GC/MS allowed the identification of ninety-one components representing 96.0 to 98.2% of the total oils. The different component contents varied according to the species. The main components were carvacrol, 1,8-cineole, α-thujone, α-terpineol and α-pinene. The PCA and the HCA of the selected essential oil components and the inhibition zone diameter (IZD) values identified four species groups and subgroups. Each essential oils group constituted a chemotype responsible for their bacterial inhibition ability. Thymus capitatus essential oil showed the strongest antibacterial activity with MBC ranging from 0.73 to 2.94 mg mL^− 1. Conclusion: Rosmarinus officinalis, Thymus capitatus, Origanum majorana and Salvia officinalis essential oils have shown promising antibacterial activities against reference bacterial strains responsible for nosocomial diseases and foodborne illnesses.

The problem of antibiotic resistance among pathogens encourages searching for novel active molecules. The aim of the research was to assay the anti-quorum sensing (anti-QS) and antibiofilm potential of Melaleuca alternifolia essential oil and its main constituent, terpinen-4-ol, to prevent the infections due to methicillin-resistant Staphylococcus aureus strains as an alternate to antibiotics. The tea tree oil (TTO) was evaluated for its potential in inhibiting QS-dependent phenomena such as violacein production in Chromobacterium violaceum, swarming motility of Pseudomonas aeruginosa PAO1, and biofilm formation in MRSA strains on glass. The results showed that terpinen-4-ol was able to inhibit MRSA strain biofilm formation on the glass strips by 73.70%. TTO inhibited the violacein production at a mean inhibitory concentration (MIC) value of 0.048 mg/mL by 69.3%. At 100 µg/mL TTO and terpinen-4-ol exhibited inhibition in swarming motility of PAO1 by 33.33% and 25%, respectively. TTO revealed anti-QS and anti-biofilm activities at very low concentrations, but it could be further investigated for new molecules useful for the treatment of MRSA infections.