Quorum sensing (QS) is a bacterial communication mechanism used to express various survival or virulence traits leading to enhanced resistance. Chromobacterium violaceum is a commonly used strain that highlights anti-QS action of bioactive substances. Here, we wanted to see if 12 selected essential oils (EO) could exert anti-QS activity. We measured the sublethal minimal QS inhibitory concentration (MQSIC) by assessing violacein production of C. violaceum along with bacterial growth. To confirm the QS disruption, we also proceed to surface bacterial observations using scanning electron microscopy (SEM). We showed that cis-cis-p-menthenolide extracted and isolated from a plant endemic to occidental Mediterranean Sea islands, Mentha suaveolens ssp. insularis, acts as an inhibitor of violacein production and biofilm formation. Measured MQSIC was much lower than the minimal inhibitory concentration (MIC): 0.10 mg·mL−1 vs. 3.00 mg·mL−1. Moreover, disturbance of QS-related traits was confirmed by the degradation of C. violaceum biofilm matrix. There is a clear structure–activity relationship between cis-cis-p-menthenolide and anti-QS activity. Indeed, its isomer molecule (mintlactone) exerts a poor anti-QS action. These results indicate that inhibition of violacein production and biofilm formation by cis-cis-p-menthenolide might be related to a disruption in the QS mechanism.
Today, an alarming rise of bacterial gastroenteritis in humans resulting from consuming Campylobacter-tainted foods is being observed. One of the solutions for mitigating this issue may be the antibacterial activity of essential oils. In the present research, we propose to study the antibacterial activity against Campylobacter and other Gram-negative bacteria of Daucus carota essential oil and its active molecules. In addition, a few chemically synthesized molecules such as (E)-methylisoeugenol, Elemicin, and eugenol were also studied. The results showed that the essential oil itself and its most active component, (E)-methylisoeugenol, exhibited bactericidal effects. Similar effects were detected using purified and chemically synthesized molecules. Also, it was observed that the Daucus carota essential oil and its active molecules affected intracellular potassium and intracellular ATP contents in Campylobacter cells. Inhibition of the membrane bound FOF1-ATPase was also observed. Eventually, for the first time, the efflux mechanism of active molecules of Daucus carota essential oil was also identified in gamma proteobacteria and its specific antibacterial activity against Campylobacter jejuni was associated with the lack of this efflux mechanism in this species.
Essential oils are known to exhibit diverse antimicrobial properties, showing their value as a natural resource. Our work aimed to investigate the primary mode of action of Cuban Lippia graveolens (Kunth) essential oil (EO) against Salmonella enterica subsp. enterica serovar Typhimurium (S. enterica ser. Typhimurium). We assessed cell integrity through various assays, including time-kill bacteriolysis, loss of cell material with absorption at 260 and 280 nm, total protein leakage, and transmission electron microscopy (TEM). The impact of L. graveolens EO on membrane depolarization was monitored and levels of intracellular and extracellular ATP were measured by fluorescence intensity. The minimum inhibitory and bactericidal concentrations (MIC and MBC) of L. graveolens EO were 0.4 and 0.8 mg/mL, respectively. This EO exhibited notable bactericidal effects on treated cells within 15 min without lysis or leakage of cellular material. TEM showed distinct alterations in cellular ultrastructure, including membrane shrinkage and cytoplasmic content redistribution. We also observed disruption of the membrane potential along with reduced intracellular and extracellular ATP concentrations. These findings show that L. graveolens EO induces the death of S. enterica ser. Typhimurium, important information that can be used to combat this foodborne disease-causing agent.
The essential oils are a powerful natural resource with antibacterial activity. This work aims at the mode of action of Lippia graveolens Kunth essential oil of plants growing in Cuba on Salmonella enterica subsp. enterica serovar Typhimurium. The effects of this oil on cell integrity were determined by time-kill, bacteriolysis and loss of 260 and 280-nm-absorbing material assays and total proteins leakage. Also, depolarization of the membrane by essential oil was monitored and intracellular and extracellular ATP was measured. The transmission electron microscopy (TEM) was used for observed morphologic change. Minimum inhibitory concentration (MIC) of L. graveolens essential oil and minimum bactericide concentration (MBC) were 0.4 and 0.8 mg/mL respectively. This essential oil showed a bactericidal action against over S . Typhimurium in a few minutes. After treatment, the cell lysis was not occurred, but little intracellular material and total proteins leakage were observed. This essential oil depolarizes the cell membrane, disturb metabolic processes and changes the structure of cytoplasmic membrane. These results suggest as primary mode of action of L. graveolens essential oil over S . Typhimurium an increased permeability of the membrane and depolarization of inside membrane, inhibition of ATPase or disturbance in proton motive force that finally provokes death of cells. L. graveolens essential oil is a botanic resource can be used for the control of the salmonellosis, foodborne disease.