Antimicrobial properties of essential oil extracted from Schizonepeta annua against methicillin-resistant Staphylococcus aureus via membrane disruption.

Schizonepeta annua (Pall.) Schischk. has long been traditionally employed in China for its anti-inflammatory, antimicrobial, and soothing properties. This study evaluates the antibacterial properties of essential oil extracted from Schizonepeta annua (SEO) and oregano (OEO) against methicillin-resistant Staphylococcus aureus (MRSA). SEO and OEO demonstrated substantial antibacterial efficacy, with SEO exhibiting significantly enhanced antibacterial activity due to its complex composition. Mechanistic investigations revealed that both essential oils disrupt bacterial membrane integrity and biosynthetic pathways, leading to the extrusion of intracellular contents. Metabolomic analyses using GC-Q-TOF-MS highlighted SEO's selective targeting of bacterial membranes, while non-targeted metabolomics indicated significant effects on MRSA's amino acid metabolism and aminoacyl-tRNA biosynthesis. These findings suggest that SEO causes considerable damage to MRSA cell membranes and affects amino acid metabolism, supporting its traditional use and highlighting its potential in treating infections. Our results offer robust theoretical support for SEO's role as an antimicrobial agent and establish a solid foundation for its practical application in combating multidrug-resistant infections.

Evaluation of the activity and mechanisms of oregano essential oil against PRV in vivo and in vitro.

BACKGROUND: The Pseudorabies Virus (PRV) leading to pseudorabies and causes huge economic losses in pig industry. The development of novel PRV variations has diminished the efficacy of traditional vaccinations, and there is yet no medication that can stop the spread of PRV infection. Therefore, PRV eradication is challenging. Oregano essential oil, the plant-based ingredient for medication feed have been shown to has strong anti-herpesvirus activity, but no anti-PRV function has been reported. RESULTS: The current study assessed the anti-pseudorabies virus (PRV) activity of oregano essential oil and explored its mechanisms and most effective components against PRV. Our in vivo findings demonstrated that oregano essential oil could decrease the PRV load in tissues, mitigate tissue lesions, and enhance the survival rate of mice. The potential antiviral mechanism involves augmenting humoral and cellular immune responses in PRV-infected mice. To further investigate the most effective components of oregano essential oil against PRV, an in vitro study was conducted, revealing that oregano essential oil and its main constituents, carvacrol and thymol, all diminished PRV intracellular proliferation in vitro. Carvacrol exhibited the most potent anti-PRV effect, serving as the primary contributor to oregano essential oil's anti-PRV activity. The mechanisms underlying carvacrol's anti-PRV properties include the upregulation of cytokines TNF-α, IFN-ß, IFN-γ, IL-12, and the inhibition of PRV-induced apoptosis in BHK-21 cells. CONCLUSIONS: Our study provides an effective drug for the prevention and control of PRV infection.

Inhibitory effect and mechanism of oregano essential oil on Listeria monocytogenes cells, toxins and biofilms.

Listeria monocytogenes (L. monocytogenes) is a prevalent foodborne pathogen with a remarkable capacity to form biofilms on utensil surfaces. The Listeriolysin O (LLO) exhibits hemolytic activity, which is responsible for causing human infections. In this study, we investigated the inhibitory effect and mechanism of oregano essential oil (OEO) on L. monocytogenes, evaluated the effects on its biofilm removal and hemolytic activity. The minimum inhibitory concentration (MIC) of OEO against L. monocytogenes was 0.03 % (v/v). L. monocytogenes was treated with OEO at 3/2 MIC for 30 min the bacteria was decreased below the detection limit (10 CFU/mL) in PBS and TSB (the initial bacterial load was about 6.5 log CFU/mL). The level of L. monocytogenes in minced pork co-cultured with OEO (15 MIC) about 2.5 log CFU/g lower than that in the untreated group. The inhibitory mechanisms of OEO against planktonic L. monocytogenes encompassed perturbation of cellular morphology, elevation in reactive oxygen species levels, augmentation of lipid oxidation extent, hyperpolarization of membrane potential, and reduction in intracellular ATP concentration. In addition, OEO reduced biofilm coverage on the surface of glass slides by 62.03 % compared with the untreated group. Meanwhile, OEO (1/8 MIC) treatment reduced the hemolytic activity of L. monocytogenes to 24.6 % compared with the positive control. Molecular docking suggested carvacrol and thymol might reduce the hemolytic activity of L. monocytogenes. The results of this study demonstrate that OEO exhibits inhibitory effects against L. monocytogenes, biofilms and LLO, which had potential as natural antimicrobial for the inhibition of L. monocytogenes.

Essential oil of oregano (Origanum vulgare L.) reduces infection and proliferation of Toxoplasma gondii in BeWo cells with induction of autophagy and death of tachyzoites through a mechanism similar to necrosis.

Toxoplasmosis poses a global health threat, ranging from asymptomatic cases to severe, potentially fatal manifestations, especially in immunocompromised individuals and congenital transmission. Prior research suggests that oregano essential oil (OEO) exhibits diverse biological effects, including antiparasitic activity against Toxoplasma gondii. Given concerns about current treatments, exploring new compounds is important. This study was to assess the toxicity of OEO on BeWo cells and T. gondii tachyzoites, as well as to evaluate its effectiveness in in vitro infection models and determine its direct action on free tachyzoites. OEO toxicity on BeWo cells and T. gondii tachyzoites was assessed by MTT and trypan blue methods, determining cytotoxic concentration (CC50), inhibitory concentration (IC50), and selectivity index (SI). Infection and proliferation indices were analyzed. Direct assessments of the parasite included reactive oxygen species (ROS) levels, mitochondrial membrane potential, necrosis, and apoptosis, as well as electron microscopy. Oregano oil exhibited low cytotoxicity on BeWo cells (CC50: 114.8 µg/mL ± 0.01) and reduced parasite viability (IC50 12.5 ± 0.06 µg/mL), demonstrating 9.18 times greater selectivity for parasites than BeWo cells. OEO treatment significantly decreased intracellular proliferation in infected cells by 84% after 24 h with 50 µg/mL. Mechanistic investigations revealed increased ROS levels, mitochondrial depolarization, and lipid droplet formation, linked to autophagy induction and plasma membrane permeabilization. These alterations, observed through electron microscopy, suggested a necrotic process confirmed by propidium iodide labeling. OEO treatment demonstrated anti-T. gondii action through cellular and metabolic change while maintaining low toxicity to trophoblastic cells.

Green manure (Crotalaria juncea L.) enhances Origanum vulgare L. biomass accumulation, essential oil yield, and phytochemical properties.

Green manure (GM) may reduce the use of chemical fertilizers, been an ecologically appropriate strategy to cultivation of medicinal plants. Crotalaria juncea, is one of the most used because it adapts to different climatic and high nitrogen content. Origanum vulgare. is widely used in cooking, pharmaceutical, cosmetic industries and food products. The objectives of this study were to evaluate the GM on biomass, essential oil (EO), phenolic and antioxidant. The experiment consisted: control; 150, 300, 450, and 600 g (Sh= leaves+steam) more 200 g roots (R); 600 g aerial part; 200 g roots; and soil with 300 g cattle manure per pot. The highest dry weights were observed in the presence of GM and cattle manure (90 days). The control had an EO production 75% lower in relation to the dose of 450 g GM (Sh+R). Principal component analysis showed that GM and cattle manure positively influenced the dry weight, content, yield, and EO constituents, and total flavonoids. The GM contributed to the accumulation of the major EO compounds (trans-sabinene hydrate, thymol, terpinen-4-ol). The GM management may be beneficial for cultivating, because it can increase the production of biomass and the active components, in addition to being an inexpensive resource.

Origanum Grosii Extracts as Novel Eco-Friendly Corrosion Inhibitors for Mild Steel In HCl Medium.

Two green inhibitors extracted from an endemic species (Origanum grosii (Og)) using two solvents of different polarity (water and ethanol), OgW (aqueous extract) and OgE (ethanolic extract), were used for the anticorrosion of mild steel (M steel) in a 1 M HCl medium. Anticorrosive performance of OgW and OgE was assessed using standard electrochemical techniques, EIS/PDP measurements, weight loss method and SEM/EDX surface analysis. The results show that OgW achieves a maximum inhibition efficiency of 92 % and that the extract in aqueous medium (more polar) is more efficient than the extract in ethanolic medium (less polar). Both extracts act as mixed inhibitors and their corrosion process is predominantly governed by a charge transfer. Concentration and temperature effect was studied and shown that they are two antagonistic parameters for the evolution of inhibitory effectiveness of both OgW and OgE. The adsorption isotherms of the two inhibitors OgE and OgW obey to the Langmuir adsorption model. Moreover, the examination of SEM images and EDX spectra support a deposit of both extracts on the metal surface by an adsorption phenomenon. Besides, theoretical approach of the molecular structures of the major compounds M-OgW and M-OgE and inhibition efficiency was examined via DFT calculations and molecular dynamics simulations and it was consistent with the experimental findings.

Evaluation of Chemical Profile and Biological Properties of Extracts of Different Origanum vulgare Cultivars Growing in Poland.

This research studied the phenolic content compared with the antioxidant properties of various O. vulgare (Lamiaceae) cultivars grown in Poland. The research results in this paper indicate that the dominant ingredient in all oregano cultivars was rosmarinic acid, known for its strong antioxidant properties. The highest amounts of rosmarinic acid (87.16 ± 4.03 mg/g dm) were identified in the O. vulgare spp. hirtum (Link) Ietsw. Other metabolites identified in the studied extracts include luteolin O-di-glucuronide-O-di-pentoside (30.79 ± 0.38 mg/g dm in the 'Aureum' cultivar), 4'-O-glucopyranosyl-3', 4'-dihydroxy benzyl-protocatechuate (19.84 ± 0.60 mg/g dm in the 'Margerita' cultivar), and p-coumaroyl-triacetyl-hexoside (25.44 ± 0.18 mg/g dm in the 'Margerita' cultivar). 'Hot & spicy' and 'Margerita' cultivars were characterized by the highest activity in eliminating OH• and O2•- radicals. Extracts from Greek oregano had the highest ability to scavenge DPPH radicals and chelate iron ions. This research has also provided new evidence that oregano has anti-migratory, cytotoxic properties and influences the viability of gastric cancer cells (the highest cytotoxicity was attributed to the 'Hot & spicy' cultivar, which performed the worst in antioxidant properties tests). Extracts from the tested cultivars at a concentration of 0.625% effectively inhibited the growth of S. aureus and P. aeruginosa bacteria. It seems that the oregano grown in Poland is of good quality and can be successfully grown on a large scale if the appropriate use is found.

Chemical and Bioactive Evaluation of Essential Oils from Edible and Aromatic Mediterranean Lamiaceae Plants.

The Lamiaceae family, which includes several well-known aromatic plants, is scientifically relevant due to its essential oils (EOs). In this work, four EOs from Mediterranean species, namely Origanum vulgare L., Rosmarinus officinalis L., Salvia officinalis L., and Thymus vulgaris L., were evaluated for their volatile profiles and the biological activity in vitro to assess their potential use in the food and cosmetic sector. GC/MS analysis revealed dominant compounds, such as carvacrol, thymol, and eucalyptol. Regarding biological action, the samples exhibited antioxidant, cytotoxic, anti-inflammatory, antimicrobial, and antifungal activities, with O. vulgare and T. officinalis standing out. T. vulgaris showed the lowest EC50 in the reducing power assay, and O. vulgare had the lowest EC50 in the DPPH assay. Most EOs also displayed excellent anti-inflammatory responses and antifungal properties, with O. vulgare and T. vulgaris also demonstrating antibacterial activity. All EOs from Mediterranean species showed cytotoxicity against tumoral cell lines. Overall, the selected EOs stood out for their interesting bioactivities, with the obtained results underscoring their potential as natural preservatives and bioactive agents in various industrial applications, including food, pharmaceuticals, and cosmetics.

Activity of Common Thyme (Thymus vulgaris L.), Greek Oregano (Origanum vulgare L. ssp. hirtum), and Common Oregano (Origanum vulgare L. ssp. vulgare) Essential Oils against Selected Phytopathogens.

The aim of this study was to determine the activity of common thyme (Thymus vulgare L.), Greek oregano (Origanum vulgare L. ssp. hirtum), and common oregano (Origanum vulgare L. ssp. vulgare) essential oils (EOs) against selected phytopathogenic microorganisms in relation to their chemical profile. The EOs were obtained from the herbs of 2-year-old plants cultivated in the organic farming system in a temperate climate in Central Europe. The EOs' composition was determined by GC/MS and GC/FID. The investigated species were represented by the following three chemotypes: 'thymol' for common thyme, 'carvacrol' for Greek oregano, and mixed 'caryophyllene oxide + ß-caryophyllene' for common oregano. The antimicrobial activity of the EOs was assessed based on minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) values. The plant pathogenic bacteria Pseudomonas syringae, Xanthomonas hortorum, Erwinia carotovora, and fungi: Fusarium culmorum, Alternaria alternata, Botrytis cinerea, Epicoccum purpurascens, Cladosporium cladosporioides, Phoma strasseri, and Pythium debaryanum were tested. The EOs revealed a stronger inhibitory effect against fungal growth in comparison to bacterial growth (MIC: 0.016-2 µL/mL for fungi and 0.125-4 µL/mL for bacteria). Common thyme and Greek oregano EOs indicated stronger antimicrobial power than common oregano EO. These results were associated with the chemical profile of the analysed EOs. The growth of examined bacteria and fungi strains (in particular, X. hortorum, F. culmorum, and P. debaryanum) were negatively correlated with the content of phenolic monoterpenes and monoterpene hydrocarbons. Among the tested strains, P. strasseri turned out to be the most sensitive (MIC 0.016 µL/mL) and E. carotovora the most resistant (MIC 0.250-4 µL/mL) to all investigated EOs.

Essential Oil-Based Soap with Clove and Oregano: A Promising Antifungal and Antibacterial Alternative against Multidrug-Resistant Microorganisms.

The transmission of microorganisms via hands is a critical factor in healthcare-associated infections (HAIs), underscoring the importance of rigorous hand hygiene. The rise of antimicrobial-resistant microorganisms, driven in part by the overuse of antibiotics in clinical medicine, presents a significant global health challenge. Antimicrobial soaps, although commonly used, may exacerbate bacterial resistance and disrupt skin microbiota, posing additional health risks and environmental hazards. Essential oils, with their broad-spectrum antimicrobial properties, offer a promising alternative. This study evaluates the antimicrobial activity of essential oils against various bacterial and fungal strains, including multidrug-resistant isolates. Using a range of in vitro and in vivo antimicrobial assays, including minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), and minimal fungicidal concentration (MFC), the essential oils were tested against a broad spectrum of pathogens. Additionally, the chemical composition of the oils was analyzed in detail using gas chromatography-mass spectrometry (CG-MS). Clove, oregano, and thyme oils demonstrated potent inhibition of all tested ATCC bacterial strains, with MIC values ranging from 3.125 to 50 µL/mL. These oils also showed significant activity against multidrug-resistant Escherichia coli and Pseudomonas aeruginosa strains. Notably, clove oil exhibited remarkable efficacy against fungal strains such as Aspergillus fumigatus and Trichophyton rubrum, with MIC values as low as 1.56 µL/mL. Synergy tests revealed that combinations of clove, oregano, and thyme oils yielded significantly lower MIC values than individual oils, indicating additive or synergistic effects. The formulation of a soap incorporating clove and oregano oils demonstrated efficacy comparable to synthetic antiseptics in vivo. These findings highlight the exceptional antimicrobial potential of essential oils, mainly clove and oregano, against resistant microorganisms, offering a viable alternative to conventional antimicrobial agents.

Effects of an essential oil blend rich in cinnamaldehyde and carvacrol on rumen biohydrogenation and fatty acid profile in the longissimus lumborum of growing lambs.

BACKGROUND: Essential oils extracted from cinnamon bark and oregano are rich in cinnamaldehyde and carvacrol and show potential for promoting animal performance. However, their impact on rumen biohydrogenation and the fatty acid composition of meat has not been reported. The hypothesis of this study was that a blend of essential oils rich in cinnamaldehyde and carvacrol would inhibit rumen biohydrogenation and promote the accumulation of polyunsaturated fatty acids (PUFAs) in lamb meat. The present study evaluated the effect of a blend essential oil (EO) rich in cinnamaldehyde and carvacrol on the nutrient digestibility, rumen biohydrogenation, growth performance, and fatty acid profile of the longissimus lumborum of lambs. RESULTS: Sixty male lambs with an average age of 84 ± 0.98 days and initial body mass of 25.4 ± 0.29 kg (mean ± standard deviation) were assigned randomly to four diets, and supplemented with 0 (EO0), 30 (EO30), 60 (EO60), and 120 (EO120) mg kg-1 dry matter of EO for 60 days. Although dry matter and neutral detergent fiber digestibility all showed a linear decrease (P ≤ 0.02) with increasing quantities of EO, final body mass and average daily gain increased linearly (P = 0.04), and average daily weight gain (ADG)/dry matter intake (DMI) tended to increase linearly (P = 0.07). Increasing EO supplementation resulted in a linear decrease in total volatile fatty acid concentration, acetate molar percentage, and acetate-to-propionate ratio (P ≤ 0.03), with the EO120 treatment being lower than the other EO treatments (P ≤ 0.05). Seven lambs from the EO120 treatment and seven lambs from the EO0 treatment were randomly slaughtered. It was observed that the proportions of C18:2n6c and PUFA in longissimus lumborum were higher in the EO120 treatment than the EO0 treatment (P ≤ 0.05). The relative abundance of Firmicutes in the rumen was decreased by the EO120 treatment in comparison with the EO0 treatment (P ≤ 0.05). Furthermore, the predicted relative abundances of genes encoding for conjugated linoleic acid reductase tended to decrease with the EO120 treatment (P = 0.06). CONCLUSIONS: We demonstrated that supplementation of the EO blend rich in cinnamaldehyde and carvacrol can enhance lamb growth performance and promote the deposition of desirable PUFAs in meat. © 2024 Society of Chemical Industry.

The impact of dietary oregano essential oil supplementation on fatty acid composition and lipid stability in eggs stored at room temperature.

*1. In many countries, eggs are not refrigerated and must be stored at room temperature. The objective of this study was to explore the effects of dietary oregano oil (275 mg/kg; ORE) versus an unsupplemented control diet (CON) on laying hens on the shelf life and fatty acid profile of eggs.2. Treatments were randomly distributed into 10 pens containing 27 birds each. A total of 200 eggs were collected from both groups on the same day and were stored for either 0, 10, 21 and 35 d. At each storage time, egg yolks were analysed for fatty acid profile and lipid peroxidation.3. The main indicator of lipid peroxidation, malondialdehyde (MDA), was significantly lower in ORE eggs compared to CON eggs (p = 0.001). Storage time had a significant impact on MDA concentrations (p = 0.023), with the highest found after 35 d. Significant differences were found for individual fatty acids, saturated (SFA), monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA). Palmitic acid, stearic acid, oleic acid, linoleic acid and arachidonic acid were significantly lower in ORE eggs compared to CON eggs (p < 0.05). Palmitoleic acid (p = 0.002), linolenic acid (p = 0.001) and docosahexaenoic acid (DHA, p = 0.001) were significantly higher in ORE eggs.4. Storage only affected oleic, linolenic, linoleic, arachidonic and docosahexaenoic acids (p < 0.05). Total SFA, MUFA, n-6 and ratio of n-3 to n-6 (n-3:n-6) PUFA were significantly higher in CON eggs (p < 0.05). The ratio of SFA to PUFA (SFA:PUFA, p = 0.005) and total n-3 PUFA (p = 0.001) were significantly higher in ORE eggs.5. The n-3:n-6 ratio was significantly impacted by treatment (p = 0.021) and storage (p = 0.031) with no significant interaction. This ratio is important for human health indication and could lead to the development of designer eggs.

Food safety control in poultry industry: prevalence and antimicrobial susceptibility of Escherichia coli isolated from raw chicken and the potential use of Origanum essential oils as alternative to antibiotics.

1. The extensive use of antimicrobials in poultry production may contribute to the emergence of resistant bacteria. This study was conducted to determine the prevalence and resistance of different E. coli strains isolated from raw chicken meat and to investigate the possibility to use Lebanese native oregano essential oils as alternatives.2. In total, 250 chickens from Lebanese markets were examined for the presence of E. coli. Isolates were then screened for susceptibility using 19 antibiotics and two essential oils extracted from oregano plants.3. Of the 250 chickens tested, 80% were contaminated with E. coli. Main resistance was seen against amoxycillin, ampicillin, penicillin, tetracycline, tylosin, streptomycin and erythromycin. The highest rate of sensitivity was found in 86.1% of strains to Amoxycillin/Clavulanic acid, 80.09% to Tilmicosin. Both essential oils from Origanum syriacum (98%) and O. ehrenbergii (97.3%) showed promising potential in inhibiting the growth of the tested bacteria. Oil from O. syriacum exhibited superior efficacy against 200 E. coli strains, inhibiting 46.1% at 200 mg/l and all at 400 mg/l, while O. ehrenbergii oil showed slightly lower inhibition, affecting 41.6% at 200 mg/l and all at 400 mg/l.

Modulatory effect of Origanum vulgare essential oil and carvacrol on Malassezia spp. virulence factors.

Malassezia sympodialis and Malassezia furfur are recognized as an important part of the human and animals healthy skin microbiota, but also as an opportunistic fungus due to their association with a broad spectrum of skin and systemic infections. Human skin infections associated with Malassezia spp. are often chronic, recurrent, and topical or oral azole treatments are challenging with not always successful outcomes. A wide inter and intraspecies antifungal susceptibility variability that may suggest a combination of wild-type strains and resistant mutants was reported. The essential oils are complex hydrophobic mixtures of different compounds extracted from plants by distillation and have demonstrated antimicrobial activity. In this study, toxicity in the larvae model and the ability of Origanum vulgare essential oil (OVEO) and carvacrol to inhibit virulence factors such as hydrophobicity, adherence, and biofilm formation of M. sympodialis and M. furfur was evaluated. We have demonstrated the antifungal activity of OVEO and carvacrol against both species. Carvacrol was more active and less toxic than OVEO at low concentrations (< 1 mg/ml). Malassezia sympodialis showed a significant decrease in adherence and its ability to form biofilms when OVEO or carvacrol is present. Both Malassezia species showed reduced hydrophobicity in the presence of carvacrol. Further studies will have to be carried out to understand how these substances act, but the inhibition of some virulence factors could represent a new target to overcome the problem of drug resistance.

The work reports the research on the effect of the Origanum vulgare essential oil and carvacrol, against virulence factors such as hydrophobicity, adherence, and biofilm formation of two species of Malassezia. The inhibition of virulence factors could represent a new target to overcome the drug resistance reported.

Unravelling the Chemotaxonomic Identity of "White" and "Black" Oregano (Origanum vulgare) in Northern Greece.

The two types of oregano used by the inhabitants of the villages of Μount Belles (GR1260001), the "white" oregano with white flowers and "black" oregano with purple flowers, were studied. The two oregano types were collected from five localities along an altitudinal gradient from 217 m up to 1299 m. "White" oregano, was found in the three lowland regions (up to 752 m) where the Pannonian-Balkanic turkey oak-sessile oak forest habitat (code 91M0) dominates. The "black" oregano was collected from the two higher altitudes, at 1177 m and 1299 m, where the Asperulo-Fagetum beech forest habitat (9130) occurs. Measurements of the density and size of peltate glandular hairs (sessile glands) on calyces, bracts, and leaves suggest that "white" oregano is distinguished by its conspicuous - apparently larger - glands. These differences were reflected in the total essential oil content, with the "white" oregano being much richer (up to 4.3 mL/100 g dry weight) than the "black" (up to 0.6%). Striking differences have also been found in the volatile fraction of their essential oil composition. The "white" oregano oils were characterized by the high content of carvacrol (up to 92.6% of identified peaks, by Headspace GC-MS). On the other hand, the two "black" oregano oils have a different aromatic profile, first reported from Greece, with main components including the sesquiterpenes ß-caryophyllene, D-germacrene, δ-cadinene and ß-bisabolene. The results so far indicate that "white" and "black" oregano, Origanum vulgare subsp. hirtum and subsp. vulgare respectively, can be clearly distinguished either by their morphological (glandular) differences or by chemical (essential oil) composition.

In Vitro and In Silico Evaluation of ACE2 and LOX Inhibitory Activity of Origanum Essential Oils and Carvacrol.

Origanum spp. are used both for culinary purposes and for their biological activities. In this study, commercial Origanum majorana, Origanum minutiflorum, Origanum vulgare, and Origanum onites essential oils and their prominent constituent carvacrol were evaluated for their in vitro and in silico angiotensin-converting enzyme 2 and lipoxygenase enzyme inhibitory potentials. The essential oils were analysed by gas chromatography-flame ionisation detection and gas chromatography-mass spectrometry, where carvacrol was identified as the major component (62 - 81%), confirming the quality. In vitro enzyme inhibition assays were conducted both with the essential oils (20 µg/mL) and with carvacrol (5 µg/mL). The comparative values of angiotensin-converting enzyme 2 percent inhibition for O. majorana, O. minutiflorum, O. vulgare, and O. onites essential oils were determined as 85.5, 79.1, 74.3, and 42.8%, respectively. As a result of the enzyme assays, carvacrol showed 90.7% in vitro angiotensin-converting enzyme 2 inhibitory activity. The in vitro lipoxygenase inhibition of the essential oils (in the same order) was 89.4, 78.9, 81.1, and 73.5%, respectively, where carvacrol showed 74.8% inhibition. In addition, protein-ligand docking and interaction profiling was used to gain structural and mechanistic insights into the angiotensin-converting enzyme 2 and lipoxygenase inhibitory potentials of major Origanum essential oil constituents. The in silico findings agreed with the significant enzyme inhibition activity observed in vitro. Further in vivo studies are suggested to confirm the safety and efficacy of the oils.

Antiproliferative Effect of Essential Oil Obtained from Oregano (Lippia palmeri S. Watson) Leaves Grown in Hydroponics and LED Light.

Nowadays, light-emitting diodes (LED) provide an alternative source to sunlight with specific intensity and wavelength that promotes plant growth. The features offered by LED could also stimulate the production of secondary metabolites of pharmaceutical interest. This work analyzed the cultivation of oregano (Lippia palmeri S. Watson) in a floating root hydroponic system supplemented by full-spectrum LED artificial light. Growth indicators like height, diameter, number of shoots, and leaf length and width were measured. The essential oil (EO) composition from the leaves of wild and hydroponic conditions found thymol (41.8 %) as the main product for the former and carvacrol (47 %) in hydroponics. The antiproliferative activity of EOs on human colorectal cancer HCT-15 shows that 6.4 µg/ml for hydroponic and 7.4 µg/ml for the wild plant reduce more than 50 % the cell viability. Overall, this study indicates that hydroponic conditions and full spectrum LED modifies the composition of the EO of L. palmeri on compared with the wild plant, which effectively induces cell growth inhibition in human colorectal cancer.

Antimicrobial and Antibiofilm Efficacy and Mechanism of Oregano Essential Oil Against Shigella flexneri.

The aim of this study was to assess the antimicrobial activity of oregano essential oil (OEO) against Shigella flexneri and eradication efficacy of OEO on biofilm. The results showed that the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of OEO against S. flexneri were 0.02% (v/v) and 0.04% (v/v), respectively. OEO effectively killed S. flexneri in Luria-Bertani (LB) broth and contaminated minced pork (the initial population of S. flexneri was about 7.0 log CFU/mL or 7.2 log CFU/g), and after treatment with OEO at 2 MIC in LB broth or at 15 MIC in minced pork, the population of S. flexneri decreased to an undetectable level after 2 or 9 h, respectively. OEO increased intracellular reactive oxygen species concentration, destroyed cell membrane, changed cell morphology, decreased intracellular ATP concentration, caused cell membrane depolarization, and destroyed proteins or inhibited proteins synthesis of S. flexneri. In addition, OEO effectively eradicated the biofilm of S. flexneri by effectively inactivating S. flexneri in mature biofilm, destroying the three-dimensional structure, and reducing exopolysaccharide biomass of S. flexneri. In conclusion, OEO exerts its antimicrobial action effectively and also has a valid scavenging effect on the biofilm of S. flexneri. These findings suggest that OEO has the potential to be used as a natural antibacterial and antibiofilm material in the control of S. flexneri in meat product supply chain, thereby preventing meat-associated infections.

A GC-MS and LC-HRMS perspective on the chemotaxonomic investigation of the natural hybrid Origanum × lirium and its parents, O. vulgare subsp. hirtum and O. scabrum.

INTRODUCTION: The genus Origanum L. (Lamiaceae) is widespread in the Mediterranean region. However, approximately 75% of the species are only encountered in the eastern part. Out of these, a total of nine species (11 taxa) and three natural hybrids occur in Greece. Nevertheless, so far, there is no consensus regarding their precise botanical classification in the literature. In fact, the taxon Origanum × lirium has been proposed both as a separate species as well as natural hybrid between Origanum vulgare subsp. hirtum and Origanum scabrum. OBJECTIVES: In this scope, the aim of the current study is to shed light on the matter through the investigation of the chemical composition of both the essential oils and the polar extracts of the mentioned taxa, collected from different geographical regions of Greece. RESULTS: As it was demonstrated by both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) data, and highlighted by our comparative analysis, it can be stipulated that Origanum × lirium shares its chemotype to a large extent with its parent species concerning both volatile and polar constituents. Additionally, geographical origin conditions stood out as a key factor influencing their chemical composition. CONCLUSION: Altogether, the present work provides useful information on the chemical composition of the taxa under investigation, while our findings support the opinion that Origanum × lirium should be considered not as a separate species, but rather as a hybrid on the way to becoming a species.

µ-Raman Determination of Essential Oils' Constituents from Distillates and Leaf Glands of Origanum Plants.

A novel, inexpensive and simple experimental setup for collecting µ-Raman spectra of volatile liquids in very small quantities was developed. It takes advantage of capillary forces to detain minute volatile liquid volumes. Spectra of volatile and even scattering or absorbing media can be measured more effectively. The method is used to facilitate the collection of intensity-consistent Raman spectra from a series of reference compounds present in Origanum essential oils, in order to quantify their constituents by multiple linear regression. Wild grown Origanum plants, collected from five different regions in Greece and taxonomically identified as O. onites, O. vulgare subsp. hirtum and O. vulgare subsp. vulgare, were appropriately distilled to acquire their essential oils. Comparison of the Raman results with those from headspace gas chromatography-mass spectrometry (HS GC-MS) confirmed the successful relative quantification of the most abundant essential oil constituents, highlighting the similarities and differences of the three Origanum taxa examined. Finally, it is demonstrated that directly measuring the leaf peltate glandular hairs yields exploitable results to identify the main components of the essential oil they contain, underlining the potential of in situ (field or industry) measurements utilizing microscope-equipped portable Raman spectrometers.