Carvacrol Content Research Articles

Protective Effect of Salvia cadmica on Fibroblast Cells from t-bhp-induced Oxidative Damage

Background:: Salvia species known as "Sage" are among the important aromatic plants used in the world. This study, it was investigated the antioxidant capacity of Salvia cadmica and to investigate its protective effect on oxidative damage in t-BHP-induced fibroblast cells. Methods:: Antioxidant activity and phenolic characterization of the extract were evaluated using DPPH, TPC, TFC, FRAP, and HPLC, respectively.TAS, TOS, MDA and 8-oxo-guanine, CAT, SOD, and GPx values were examined using enzyme-linked immunosorbent assay (ELISA). The antiproliferative and apoptosis effects of Salvia cadmica ethanolic extract were determined using XTT assay and fluorescent probes in fibroblast cells. Results:: As a result of GC-MS analysis of Salvia cadmica ethanolic extract, carvacrol content was found to be high. The IC50 value of the DPPH antioxidant assay of the Salvia cadmica ethanolic extract was 80 ± 0.51 μg/mL. TPC, TFC, and FRAP values were found to be 18.25 ± 0.64 (mg gallic acid/g powder), 1.691±0.314 (mg quercetin /g powder), 31.5 ± 0.10 (mg Trolox/g powder), respectively. Total antioxidant and TOS values were found to be 0.383±0.033 (mmol Trolox Equ L-1), 16.31±0.71 (μmol H2O2 L-1) for 0.25 mg/mL, and 0.725±0.05 (mmol Trolox Equ L-1), 12.02 ±0.56 (μmol H2O2 L-1) for 0.5 mg/mL. In addition, while CAT and GPx significantly decreased enzyme activities, no significant change was observed in SOD enzyme activity. Ethanolic Salvia cadmica extract exhibited apoptotic features compared to only the t-BHP group. Conclusion:: These results suggest that Salvica cadmica extract works through a free radical mechanism.

Nutritional and Physiological Properties of Thymbra spicata: In Vitro Study Using Fecal Fermentation and Intestinal Integrity Models.

(Poly)phenolic-rich Mediterranean plants such as Thymbra spicata have been associated with several health-promoting effects. The nutritional value, as well as physiological interaction of T. spicata with the gastrointestinal tract, has not been investigated before. The nutritional composition of T. spicata leaves was here characterized by standard analytical methods. T. spicata leaves were subjected to ethanolic extraction, simulated gastrointestinal digestion, and anaerobic microbial gut fermentation. Phenols/flavonoid contents and radical scavenging activity were assessed by colorimetric methods. The volatile organic compounds (VOCs) were detected by gas chromatography coupled with mass spectrometry. The effect on intestinal integrity was evaluated using a Caco-2 monolayers mounted in a Ussing chamber. T. spicata contains a high amount of fiber (12.3%) and unsaturated fatty acids (76% of total fat). A positive change in VOCs including short-chain fatty acids was observed without significant change in viable microbe. T. spicata and carvacrol (main phenolic compound) enhanced ionic currents in a concentration-dependent manner without compromising the Caco-2 monolayer's integrity. These effects were partially lost upon simulated digestion and completely abolished after colonic fermentation in line with polyphenols and carvacrol content. Conclusion: T. spicata represents a promising nutrient for the modulation of gut microbiota and the gut barrier. Further studies must better define its mechanisms of action.

Effective Inhibition of Listeria monocytogenes Biofilm Formation by Satureja rechingeri Essential Oil: Mechanisms and Implications.

Biofilm formation by foodborne pathogens, particularly Listeria monocytogenes, poses a significant challenge in food industry facilities. In this study, we investigated the inhibitory potential of Satureja rechingeri essential oil (Sr-EO) against L. monocytogenes growth and biofilm formation. Gas chromatography-mass spectrometry analysis revealed a high carvacrol content in Sr-EO, a compound with known antimicrobial properties. We examined the effects of Sr-EO on initial attachment and preformed biofilms, using crystal violet and MTT assays to quantify attached biomass and metabolic activity, respectively. Our results demonstrated that Sr-EO not only prevented initial attachment but also effectively disrupted preformed biofilms, indicating its potential as a biofilm-control agent. Microscopy analysis revealed alterations in bacterial cell membranes upon Sr-EO treatment, leading to increased permeability and cell death. Additionally, Sr-EO significantly suppressed bacterial motility, with concentrations exceeding 0.25μL/mL completely inhibiting motility. Furthermore, gene expression analysis revealed the down regulation of genes associated with biofilm formation, attachment, and quorum sensing, suggesting that Sr-EO modulates bacterial gene transcription. These findings suggest that Sr-EO can be a promising candidate for controlling biofilm formation and bacterial contamination in food processing environments.

Anti-Cancer Study of Carvacrol in Hypoxic-Induced Colorectal Cancer Cell

Introduction: Colorectal cancer presents a complex global health challenge, characterized by complications arising from metastasis development due to rapid proliferation, adaptation to hypoxia, and angiogenesis. Carvacrol, a natural monoterpenoid phenol derived from various plants, has interest for its diverse pharmacological properties, particularly its antitumor effects. Methods: We aimed to assess the inhibitory effect of carvacrol on cell migration and proliferation in the hypoxic colorectal cancer cell line (SW480). Chemical induction of hypoxia using cobalt chloride and serially diluted concentrations of carvacrol (400, 200, 100, 50, 25, and 12.5 µg/ml) were employed to evaluate the cytotoxic effect via the MTT assay. Smaller concentrations (low IC50) were used to examine the impact of carvacrol on SW480 cell migration through a cell migration assay (50, 25, 12.5, and 6.25 µg/ml). Results: The results demonstrated a significant, dose-dependent reduction in both cell proliferation and migration with carvacrol doses. Conclusion: The findings suggest that carvacrol could be useful as an adjuvant therapy and a promising addition for patients with highly metastatic colorectal cancer.

Carvacrol in ovo delivery optimization and flow dynamics in broiler chicken eggs

In ovo delivery of carvacrol, the primary active compound in oregano essential oil (OEO) has the potential to enhance gut development in broilers. This study aimed to optimize in ovo application of OEO by investigating day and site of injection and delivery of carvacrol to different embryonic tissues. In Experiment 1, 2 d of injection (embryonic day (E) 12 or 17.5) and 3 sites of injection for OEO (air cell, amniotic fluid, or yolk) were evaluated based on hatchability and posthatching performance. Experiment 2 aimed to examine the impact of combining OEO with the nonionic surfactant polysorbate 80 (p80) at ratios to carvacrol of 0:0, 0:1, 0.5:1, and 1:1 on carvacrol concentration in amniotic fluid, blood, and yolk. The concentration of carvacrol was measured at 3, 6, and 9 h after OEO injection either without (0:1) or with (1:1) p80. Injection of OEO on E12 led to a significant lower hatchability compared to E17.5 (P ≤ 0.01; Δ = 9.2%). Injecting OEO into the air cell, amniotic fluid, or yolk at E17.5 did not significantly affect hatchability and posthatching performance. The highest concentrations of carvacrol found in egg tissues were observed when injected together with surfactant at the 1:1 ratio (P ≤ 0.001; 14.45 µM, 16.64 µM, and 124.82 µM, for air cell, amniotic fluid, and yolk, respectively) compared to the 0:0, 0:1 or 0.5:1 ratios. Carvacrol was highest in the amniotic fluid and blood at the first time point (3 h postinjection) and decreased afterward (P ≤ 0.001), whereas the concentration in yolk remained elevated up to 9 h postinjection. In conclusion, the optimization of the in ovo delivery of carvacrol resulted in that early injection (E12) had negative effects on hatchability and should be avoided. The findings also suggest that using a nonionic surfactant was crucial for an effective delivery of carvacrol in ovo and the migration from amniotic fluid to yolk within 3 h. In addition, carvacrol's persistence in yolk may serve as a route for delivery into the gastrointestinal tract via the yolk stalk during the peri-hatching phase, potentially influencing gut development.

Adaptive evolution of Salmonella Typhimurium LT2 exposed to carvacrol lacks a uniform pattern

Emergence of genetic variants with increased resistance/tolerance to natural antimicrobials, such as essential oils, has been previously evidenced; however, it is unknown whether mutagenesis follows a general or a specific pattern. For this purpose, we carried out four adaptive laboratory evolutions (ALE) in parallel of Salmonella enterica Typhimurium with carvacrol. After 10 evolution steps, we selected and characterized one colony from each lineage (SeCarA, SeCarB, SeCarC, and SeCarD). Phenotypic characterization of the four evolved strains revealed enhanced survival to lethal treatments; two of them (SeCarA and SeCarB) showed an increase of minimum inhibitory concentration of carvacrol and a better growth fitness in the presence of carvacrol compared to wild-type strain. Whole genome sequencing revealed 10 mutations, of which four (rrsH, sseG, wbaV, and flhA) were present in more than one strain, whereas six (nirC, fliH, lon, rob, upstream yfhP, and upstream argR) were unique to individual strains. Single-mutation genetic constructs in SeWT confirmed lon and rob as responsible for the increased resistance to carvacrol as well as to antibiotics (ampicillin, ciprofloxacin, chloramphenicol, nalidixic acid, rifampicin, tetracycline, and trimethoprim). wbaV played an important role in increased tolerance against carvacrol and chloramphenicol, and flhA in cross-tolerance to heat treatments. As a conclusion, no common phenotypical or genotypical pattern was observed in the isolated resistant variants of Salmonella Typhimurium emerged under carvacrol stress. Furthermore, the demonstration of cross-resistance against heat and antibiotics exhibited by resistant variants raises concerns regarding food safety.Key points• Stable resistant variants of Salmonella Typhimurium emerged under carvacrol stress• No common pattern of mutagenesis after cyclic exposures to carvacrol was observed• Resistant variants to carvacrol showed cross-resistance to heat and to antibioticsGraphical abstract

Evaluation of antifungal activity of natural compounds on growth and aflatoxin B1 production of Aspergillus parasiticus and Aspergillus flavus.

Aspergillus species cause broad spectrum infections especially invasive lethal infections in immunocompromised patients. This study aimed to assess the antifungal activity of plants and compounds including Aloe vera, Thyme, carvacrol, and nano-encapsulation of carvacrol on the growth and production of aflatoxin B1 production by Aspergillus parasiticus and Aspergillus flavus. Minimum inhibitory concentrations of extracts Aloe vera, Thyme, carvacrol, and nanocarvacrol, and fluconazole as a control were determined according to Clinical and Laboratory Standards Institute by serial microdilution protocol. Then, the effect of inhibitory concentrations of these compounds on the aflatoxin B1 production level was evaluated by real-time PCR and high-performance liquid chromatography. Our results indicate that the Aspergillus parasiticus and Aspergillus flavusare sensitive to selected plants and compounds. Our findings showed that the compounds are appropriate alternative candidates against growth and production of aflatoxin of Aspergillus spp.

Anti-Helicobacter pylori Effects of Thymus caramanicus Jalas Essential Oils: A New Antimicrobial Approach.

Helicobacter pylori are the principal causative factor in the etiological factors of chronic, active, or type B gastritis; peptic and duodenal ulcers; stomach carcinoma; and epithelial tissue lymphoid malignancies. It infects more than half of the population worldwide. To reduce H. pylori production, pharmacological therapy of H. pylori diseases typically involves using threefold treatment methods. However, as a result of such therapy, antimicrobial resistance is commonly developed. Alternative therapeutics for H. pylori diseases are thus of particular interest. Thyme essential oils (EOs) obtained from T. caramanicus Jalas plants in Iran were tested for antibacterial activity against H. pylori obtained from 320 poultry specimens in this investigation. Antibacterial activity was measured using inhibition zones, minimum inhibitory concentrations (MICs), and minimum bactericidal concentrations (MBCs). The impact of T. caramanicus Jalas essential oils on H. pylori isolate cagA, vacA, and babA2 gene expression was evaluated using a quantitative real-time PCR method (p < 0.05). The chemical content of these EOs varied significantly according to chromatographic examination. Thymol, carvacrol, and terpinene-4-ol are the most abundant components in these EOs. H. pylori was recognized as a Helicobacter species with a 175-bp PCR product of 16S rRNA in 20/20 (100%). According to PCR results, all 20 (100%) isolates belonged to H. pylori. The EOs inhibited H. pylori in a dose-dependent manner, with T. caramanicus Jalas being the most effective, followed by pterygium EOs in decreasing order. At 8 mg/mL of T. caramanicus Jalas EOs, IZs against H. pylori were 27.4 ± 0.42 mm, and at 8 mg/mL of pterygium, IZs against H. pylori were 1 ± 0.02. T. caramanicus Jalas essential oils were used to treat all bacteria, and the findings showed that T. caramanicus Jalas had a substantial inhibitory impact on the expression of cagA, vacA, and babA2 virulence-related genes (p < 0.05). In a dose-dependent manner, the EOs of T. caramanicus Jalas EO demonstrated a high degree of antimicrobial property against H. pylori bacteria. The most efficient EOs were those from T. caramanicus Jalas with relative concentrations of thymol and carvacrol, followed by the coumarin-dominated pterygium EO with reduced antibacterial activity.

Biologically active substances of new marjoram varieties and prospects for their use

Biologically active substances, essential oils, and plant extracts were studied obtained from new high-yield varieties of Origanum vulgare L. (Raduga, Belaya Ptitsa, Krymchanka) and Origanum tyttanthum Contsch. (Alcina) of the GSBSI “NBG – NSC RAS” breeding. It was found that the O. vulgare variety Belaya Ptitsa was distinguished by the content of essential oil, the main component of which was carvacrol. The maximum total content of phenolic substances and flavonoids was distinguished by an alcoholic extract from the O. tyttanthum variety. Neochlorogenic and rosemary acids, apigenin-6,8-di-C-glucoside, carvacrol, and thymol were identified in extracts of all the studied marjoram varieties. It is shown that the vegetable raw material O. vulgare cv “Belaya Ptica” is promising for the production of essential oil with a high content of carvacrol, and O. tyttanthum cv “ can serve as a raw source of rosemary acid.

ELECTROSPINNING OF PVP/CARVACROL/LANOLIN COMPOSITE NANOFIBERS

In this study, it was aimed to produce and characterize various concentrations of carvacrol and lanolin loaded PVP nanofibers via electrospinning. Various concentrations of carvacrol:lanolin were added to the PVP polymer solutions such as 2.5, 5, 7.5, 10, 12.5 and 15 wt %. Solution properties such as viscosity, conductivity, and surface tension were measured. In terms of characterization studies, SEM, FT-IR, TGA and DSC analysis were carried out. According to the results of the study, viscosity increased and conductivity decreased with carvacrol:lanolin concentration increasement. However, surface tension was not affected from carvacrol:lanolin concentration. Nanowebs quality improved with carvacrol:lanolin concentrations. Generally, nanofibers are quite fine, smooth, and uniform. FT-IR spectrums verified that PVP, carvacrol and lanolin exist in the structure of nanofibers chemically. DSC and TGA results demonstrated that the thermal stability of carvacrol and lanolin, which have poor thermal stability, was enhanced by incorporating them into the PVP nanofiber structures. Considering antibacterial properties of carvacrol and wound healing properties of lanolin, these composite nanofiber surfaces have a high potential for use in the biomedical field, especially as a wound dressing.

Antifungal effects of carvacrol, the main volatile compound in Origanum vulgare L. essential oil, against Aspergillus flavus in postharvest wheat

Plant volatile organic compounds (VOCs) with antimicrobial activity could potentially be extremely useful fumigants to prevent and control the fungal decay of agricultural products postharvest. In this study, antifungal effects of volatile compounds in essential oils extracted from Origanum vulgare L. against Aspergillus flavus growth were investigated using transcriptomic and biochemical analyses. Carvacrol was identified as the major volatile constituent of the Origanum vulgare L. essential oil, accounting for 66.01 % of the total content. The minimum inhibitory concentrations of carvacrol were 0.071 and 0.18 μL/mL in gas-phase fumigation and liquid contact, respectively. Fumigation with 0.60 μL/mL of carvacrol could completely inhibit A. flavus proliferation in wheat grains with 20 % moisture, showing its potential as a biofumigant. Scanning electron microscopy revealed that carvacrol treatment caused morphological deformation of A. flavus mycelia, and the resulting increased electrolyte leakage indicates damage to the plasma membrane. Confocal laser scanning microscopy confirmed that the carvacrol treatment caused a decrease in mitochondrial membrane potential, reactive oxygen species accumulation, and DNA damage. Transcriptome analysis revealed that differentially expressed genes were mainly associated with fatty acid degradation, autophagy, peroxisomes, the tricarboxylic acid cycle, oxidative phosphorylation, and DNA replication in A. flavus mycelia exposed to carvacrol. Biochemical analyses of hydrogen peroxide and superoxide anion content, and catalase, superoxide dismutase, and glutathione S-transferase activities showed that carvacrol induced oxidative stress in A. flavus, which agreed with the transcriptome results. In summary, this study provides an experimental basis for the use of carvacrol as a promising biofumigant for the prevention of A. flavus contamination during postharvest grain storage.

Carvacrol Treatment Reduces Decay and Maintains the Postharvest Quality of Red Grape Fruits (Vitis vinifera L.) Inoculated with Alternaria alternata.

In this study, we isolated and identified pathogenic fungi from the naturally occurring fruits of red grapes, studied their biological characteristics, screened fifteen essential oil components to find the best natural antibacterial agent with the strongest inhibitory effect, and then compared the incidence of postharvest diseases and storage potential of red grapes treated with two concentrations (0.5 EC50/EC50) of essential oil components (inoculated with pathogenic fungi) during storage for 12 d at room temperature. In our research, Alternaria alternata was the primary pathogenic fungus of red grapes. Specifically, red grapes became infected which caused diseases, regardless of whether they were inoculated with Alternaria alternata in an injured or uninjured state. Our findings demonstrated that the following conditions were ideal for Alternaria alternata mycelial development and spore germination: BSA medium, D-maltose, ammonium nitrate, 28 °C, pH 6, and exposure to light. For the best Alternaria alternata spore production, OA medium, mannitol, urea, 34 °C, pH 9, and dark conditions were advised. Furthermore, with an EC50 value of 36.71 μg/mL, carvacrol demonstrated the highest inhibitory impact on Alternaria alternata among the 15 components of essential oils. In the meantime, treatment with EC50 concentration of carvacrol was found to be more effective than 0.5 EC50 concentration for controlling Alternaria alternata-induced decay disease of red grapes. The fruits exhibited remarkable improvements in the activity of defense-related enzymes, preservation of the greatest hardness and total soluble solids content, reduction in membrane lipid peroxidation in the peel, and preservation of the structural integrity of peel cells. Consequently, carvacrol was able to prevent the Alternaria alternata infestation disease that affects red grapes, and its EC50 concentration produced the greatest outcomes.

Synergistic antimicrobial activity of essential oils in combination with phage endolysin against Salmonella Typhimurium in cooked ground beef

This study was designed to evaluate the antimicrobial activity of selected essential oils (AITC, carvacrol, eugenol, and thymol) in combination with Salmonella phage-encoding endolysin (LysPB32) against Salmonella Typhimurium KCCM 40253 in vitro and in cooked ground beef. The minimum inhibitory concentrations (MICs) of AITC, carvacrol, eugenol, and thymol against S. Typhimurium were determined in the absence and presence of LysPB32 using broth microdilution assay. In vitro antimicrobial susceptibility of S. Typhimurium to the essential oils in combination with LysPB32 was evaluated at 37 °C for 24 h using time-kill curve assay. The N-phenyl-1-napthylamine (NPN) uptake and relative bacterial fitness were measured to evaluate the outer membrane permeability and reproductive success. The morphological alterations of these treated S. Typhimurium cells were observed through a holotomographic 3D microscopy. The MICs of AITC, carvacrol, eugenol, and thymol against S. Typhimurium were significantly decreased from 6.0 to 2.9 mg/mL, 3.0 to 0.8 mg/mL, 2.8 to 0.7 mg/mL, and 1.4 to 0.7 mg/mL, respectively, when treated with LysPB32 (100 μg/mL). The combinations of carvacrol, eugenol, and thymol combined with LysPB32 effectively reduced the numbers of S. Typhimurium to below the detection limit (20 CFU/mL) after 12 h incubation at 37 °C, caused by membrane disruption. The relative bacterial fitness was significantly decreased in S. Typhimurium treated with carvacrol in the absence and presence of LysPB32. The combinations of carvacrol and eugenol with LysPB32 most effectively inhibited the growth of S. Typhimurium in cooked ground beef after 24 h of storage at 37 °C (>2 log reduction) and 7 days of refrigerated storage (>2 log reduction). Therefore, this study could provide the possibility of using the combinations of essential oils and phage endolysin as synergistic antimicrobials and also pave the way for designing effective food preservatives to enhance food safety and quality.

Electrochemical sensing of monoterpene phenols in plant essential oils using a molecularly imprinted polymer dual-template sensor

Monoterpene phenols are naturally occurring aromatic compounds found in plant essential oils, contributing to their distinct flavors, fragrances, and medicinal properties. Thymol and carvacrol, major constituents of thyme and oregano essential oils, have garnered significant attention due to their diverse biological activities. Here, we propose the development of a novel double template molecular imprinted electrochemical sensor (DT-MIP sensor) for the selective and simultaneous detection of thymol and carvacrol. The sensor's performance was optimized by varying the quantity of functional monomer, cross-linker, and template molecules during MIP synthesis. The DT-MIP sensor demonstrated superior recognition properties and high sensitivity, with detection limits of 0.11 μM for carvacrol and 0.24 μM for thymol. Selectivity tests confirmed minimal interference from potential interfering agents. GC-MS analysis validated the sensor's accuracy in detecting thymol and carvacrol concentrations in thyme and oregano essential oils. The simplicity, cost-effectiveness, and portability of the DT-MIP sensor offer promising applications in food quality control, environmental monitoring, and medical diagnostics.

Inhibition of Acinetobacter baumannii Biofilm Formation by Terpenes from Oregano (Lippia graveolens) Essential Oil.

Acinetobacter baumannii is a nosocomial pathogen known for its ability to form biofilms, leading to persistent infections and antibiotic resistance. The limited effective antibiotics have encouraged the development of innovative strategies such as using essential oils and their constituents. This study evaluated the efficacy of oregano (Lippia graveolens) essential oil (OEO) and its terpene compounds, carvacrol and thymol, in inhibiting A. baumannii biofilms. These treatments showed a minimum inhibitory concentration of 0.6, 0.3, and 2.5 mg/mL and a minimum bactericidal concentration of 1.2, 0.6, and 5 mg/mL, respectively. Sub-inhibitory doses of each treatment and the OEO significantly reduced biofilm biomass and the covered area of A. baumannii biofilms as measured by fluorescence microscopy. Carvacrol at 0.15 mg/mL exhibited the most potent efficacy, achieving a remarkable 95% reduction. Sub-inhibitory concentrations of carvacrol significantly reduced the biofilm formation of A. baumannii in stainless steel surfaces by up to 1.15 log CFU/cm2 compared to untreated bacteria. The OEO and thymol exhibited reductions of 0.6 log CFU/cm2 and 0.4 log CFU/cm2, respectively, without affecting cell viability. Moreover, the terpenes inhibited twitching motility, a crucial step in biofilm establishment, with carvacrol exhibiting the highest inhibition, followed by OEO and thymol. The study provides valuable insights into the potential of terpenes as effective agents against A. baumannii biofilms, offering promising avenues for developing novel strategies to prevent persistent infections and overcome antibiotic resistance.

Enhancement effect of carvacrol on yeast inactivation by mild pressure carbon dioxide.

Saccharomyces cerevisiae is one of the common spoilage microorganisms in fruit juices. This paper investigated the influences of carvacrol on S. cerevisiae inactivation by mild pressure carbon dioxide (MPCO2). The results demonstrated that carvacrol synergistically enhanced the antifungal activity against S. cerevisiae of MPCO2. With the increase of carvacrol concentration (20-160µg/mL), CO2 pressure (1.5-3.5MPa), process temperature (20-40°C), and treatment time (15-60min), the inactivation effect of carvacrol combined with MPCO2 on S. cerevisiae was gradually increased and significantly stronger than either single treatment. In the presence of carvacrol, MPCO2 severely disordered the plasma membrane of S. cerevisiae, including the increase of membrane permeability, and the loss of membrane potential and integrity. MPCO2 and carvacrol in combination also aggravated the mitochondrial depolarization of S. cerevisiae and reduced intracellular ATP and protein content. This study suggests the potential of carvacrol and pressurized CO2 as an alternative technology for food pasteurization.

The antimicrobial substance's content influences the membrane blocking mechanism in cross-flow filtration – Flow cytometric assessment of the membrane's activity

The study aims to investigate the antibacterial properties of carvacrol-loaded cellulose acetate membranes in cross-flow filtration using an advanced microbiological flow cytometry method and analyze the influence of carvacrol's presence on the pore-blocking mechanisms. The commercial cellulose acetate microfiltration membranes were impregnated with carvacrol using supercritical carbon dioxide as a solvent and transport medium. This technique enables active substance distribution throughout the whole polymer volume on the molecular level. Carvacrol loadings of up to 30 % were obtainable, whereby the membranes preserved their microstructure and functionality for loadings of up to 25 %. The protocol for flow cytometric assessment of the cell deposition on the membrane surface during the cross-flow filtration was developed. The results showed that the presence of carvacrol considerably decreased Staphylococcus aureus deposition on the membrane (from the order of magnitude 107/(cm2membrane surface·mLpermeate) to 105/(cm2membrane surface·mLpermeate)). The membranes containing 20 % and 25 % carvacrol showed the largest permeate flux and the smallest number of deposited bacteria. The largest fraction of dead bacteria (42 %) was detected in the membrane containing 25 % carvacrol. Mathematical modeling showed that the pore-blocking mechanism changed with the increase in carvacrol content, from the cake filtration phenomenon via the intermediate blocking towards standard and complete blocking mechanisms.

Effect of Different Plant Growth-Promoting Rhizobacteria on Biological Soil Properties, Growth, Yield and Quality of Oregano (Origanum onites L.)

Intensive agriculture uses continuous chemical fertilizers to increase crop yields, but excessive use of fertilizers leads to environmental pollution, permanent changes in physicochemical conditions in soil ecology, deterioration of soil biological health, leaching of nutrients, surface and groundwater pollution and eutrophication. Plant growth-promoting rhizobacteria (PGPR) are becoming increasingly important for ensuring crop safety, increasing nutrient uptake and output, lowering fertilizer costs, preventing environmental contamination and promoting sustainable agriculture and agricultural resources. Therefore, the purpose of this study was to identify and evaluate the effects of fifteen bacteria strains that were isolated from various acidic rhizospheric soils as biofertilizers on soil biological properties. Growth, yield and quality traits were analyzed, and various PGPR were identified using 16S ribosomal RNA of Turkish oregano. Fifteen bacterial inoculations with 1-aminocyclopropane-1-carboxylate (ACC) deaminase, N2-fixing, P-solubilizing and/or IAA-producing genes were used in the experiment, which was carried out in a randomized block design with five replicates (each with three pots) and a control without inoculation. Increased biological activity in soil inoculated with bacteria with multiple traits was confirmed by high C and N content in microbial biomass, urease, dehydrogenase and acid and alkaline phosphatase activities. Essential oil content, oil yield, thymol and carvacrol contents increased by 0.5–40.1%, 5.9–71.9%, 0.07–16.7% and 0.3–9.2%, respectively, as a result of bacterial inoculation. Oil content ranged from 2.02% to 2.83%; carvacrol (66.1–72.2%) was the main constituent, followed by thymol (14.5–16.9%) and linalool (1.38–3.68%). Two large PGPR groups were formed based on genetic distance analysis. Responses were variable and depended on the inoculant strain and the parameters being evaluated. The results indicate PGPR has clear potential for improving the yield of cultivated aromatic and essential oil plants, such as oregano.

Carvacrol and Cancer Treatment: The Effect of Carvacrol in Inducing Apoptosis of Breast Cancer Cells

Background: Carvacrol is a natural monoterpene phenolic compound that has biological activities with therapeutic applications, and this study aimed to investigate the apoptotic effect of Carvacrol on the human breast cancer cell line MDA-MB-231. Materials and Methods: After the preparation and cultivation of MDA-MB-231 cells, the IC50 value of Carvacrol on the cells was evaluated by the MTT assay method, and then the induction of apoptosis in the cell line treated with different concentrations of Carvacrol was observed by DAPI staining. The qPCR method was used to check the expression levels of Bax, P53, and Bcl-2 genes at the mRNA level. The results were analyzed using GraphPad Prism 9 software. Results: The results showed that the cytotoxicity of Carvacrol against MDA-MB-231 cancer cells was dose-dependent at 24 (P=0.034) and 48 (P=0.041) hours. The IC50 of Carvacrol at 48 h was 154.2 μM. The MDA-MB-231 cells treated with Carvacrol showed induction of apoptosis from the mitochondrial pathway by increasing the expression of P53 and BAX genes and decreasing the expression of the anti-apoptotic Bcl-2 gene. In addition, the number of apoptotic cells with dense DNA increased significantly in the Carvacrol treatment group (P=0.001). Conclusion: This study showed that treatment of the MDA-MB-231 cell line with Carvacrol can inhibit its growth and proliferation through the induction of apoptosis from the BAX/BCL-2 pathway. Considering the significant inhibitory effect of the treatment of cells with the Carvacrol compound, it seems that there is a suitable research field for using this compound in the control and treatment of breast cancer.

The effect of induced polyploidy on phytochemistry, cellular organelles and the expression of genes involved in thymol and carvacrol biosynthetic pathway in thyme (Thymus vulgaris).

Induced polyploidy usually results in larger vegetative and reproductive plant organs. In order to study the effect of chromosome doubling on Thymus vulgaris, three levels of colchicine concentration including 0.1, 0.3 and 0.5% (w/v) were applied for 6, 12 and 24 hours on apical meristem of 2- and 4-leaf seedlings. Ploidy level was evaluated by flow cytometry and microscopic chromosome counting. Chemical composition of essential oils extracted by hydro-distillation was analyzed by gas Chromatography/mass spectrometry (GC/MS) and gas Chromatography (GC). The application of 0.3% colchicine at 4-leaf seedling for 6 hours resulted in the highest survival rate and the highest number of tetraploid plants. Cytogenetic and flow cytometry analyses confirmed the increase of chromosome number from 2n=2x=30 in diploids to 2n=4x=60 in induced tetraploids. Tetraploid plants had larger leaves, taller and thicker stems, dense branching, longer trichome, larger stomata, larger guard cells, and decreased number of stomata. The number of chloroplasts and mitochondria increased significantly in tetraploid plants by 1.66 and 1.63 times, respectively. The expression of CYP71D178, CYP71D180 and CYP71D181 increased in tetraploids by 3.27, 7.39 and 2.15 times, respectively, probably resulting in higher essential oil compounds, as tetraploids outyielded the diploid plants by 64.7% in essential oil, 40.9% in thymol and 18.6% in carvacrol content.