Evaluation of the anticancer activity of Origanum Marjoram as a safe natural drink for daily use.

BACKGROUND: Chemotherapeutic agents have numerous side effects. There is a major interest in using natural and safe plants as food or drink to prevent from cancer. Origanum marjoram (OMAE) is a medicinal plant that can be used as a tea, food, and additive in traditional medicine. OBJECTIVE: This study aimed to evaluate the potential anticancer effects of OMAE as a soft drink for daily use against a model cancer, prevention and treatment. METHOD: MCF-7 cells were chosen as model cancer cells. The MTT assay was used to assess the in vitro inhibitory effects of OMAE on cell growth. Moreover, quantitative real-time PCR (qRT-PCR) was used to detect specific genes associated with cancer, such as ESR1, Bax, Bcl-2, and p53. Furthermore, the DNA damage was evaluated using the comet assay. RESULTS: OMAE has IC50 of 53.1 and IC90 of 97.5 µg/ml dependent inhibition of cell proliferation after 48 h of treatment toward MCF-7. Also, a significant decrease in the expression level of the ESR1 gene in the MCF-7 cell line. Furthermore, there was a significant increase in the comet length and comet-positive cells after treatment with OMAE (88.7%) compared with those in the untreated control cells (9.5%), suggesting a high induction of DNA damage by OMAE. Also, OMAE showed a modification in bcl-2, tumor suppressor gene (p53), and Bax levels and influenced the BAX/BCL-2 ratio via releasing the cytochrome C. CONCLUSION: The results of the study were promising, suggesting that the reduced apoptotic rate of MCF-7 breast cancer cells in this work was correlated to the potential anticancer effect of OMAE which would be a suitable preventable drink against cancer. However, further studies are needed to fully understand the potential of OMAE as a cancer treatment.

Intraspecific divergence in essential oil content, composition and genes expression patterns of monoterpene synthesis in Origanum vulgare subsp. vulgare and subsp. gracile under salinity stress.

BACKGROUND: Oregano (Origanum vulgare L.), one of the important medicinal plants in the world, has valuable pharmacological compounds with antimicrobial, antiviral, antioxidant, anti-inflammatory, antispasmodic, antiurolithic, antiproliferative and neuroprotective activities. Phenolic monoterpenes such as thymol and carvacrol with many medical importance are found in Oregano essential oil. The biosynthesis of these compounds is carried out through the methyl erythritol-4 phosphate (MEP) pathway. Environmental stresses such as salinity might improve the secondary metabolites in medicinal plants. The influence of salinity stress (0 (control), 25, 50 and 100 mM NaCl) on the essential oil content, composition and expression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), γ-terpinene synthase (Ovtps2) and cytochrome P450 monooxygenases (CYP71D180) genes involved in thymol and carvacrol biosynthesis, was investigated in two oregano subspecies (vulgare and gracile). RESULTS: Essential oil content was increased at low NaCl concentration (25 mM) compared with non-stress conditions, whereas it was decreased as salinity stress intensified (50 and 100 mM). Essential oil content was significantly higher in subsp. gracile than subsp. vulgare. The highest (0.20 mL pot-1) and lowest (0.06 mL pot-1) amount of essential oil yield was obtained in subsp. gracile at 25 and 100 mM NaCl, respectively. The content of carvacrol, as the main component of essential oil, decreased with increasing salinity level in subsp. gracile, but increased in subsp. vulgare. The highest expression of DXR, Ovtps2 and CYP71D180 genes was observed at 50 mM NaCl in subsp. vulgare. While, in subsp. gracile, the expression of the mentioned genes decreased with increasing salinity levels. A positive correlation was obtained between the expression of DXR, Ovtps2 and CYP71D180 genes with carvacrol content in both subspecies. On the other hand, a negative correlation was found between the expression of CYP71D180 and carvacrol content in subsp. gracile. CONCLUSIONS: The findings of this study demonstrated that both oregano subspecies can tolerate NaCl salinity up to 50 mM without significant reduction in essential oil yield. Also, moderate salinity stress (50 mM NaCl) in subsp. vulgare might increase the carvacrol content partly via increment the expression levels of DXR, Ovtps2 and CYP71D180 genes.

Origanum majorana L. protects against neuroinflammation-mediated cognitive impairment: a phyto-pharmacological study.

BACKGROUND: Neuroinflammation and oxidative stress are critical players in the pathogenesis of numerous neurodegenerative diseases, such as Alzheimer's disease (AD) which is responsible for most cases of dementia in the elderly. With the lack of curative treatments, natural phenolics are potential candidates to delay the onset and progression of such age-related disorders due to their potent antioxidant and anti-inflammatory effects. This study aims at assessing the phytochemical characteristics of Origanum majorana L. (OM) hydroalcohol extract and its neuroprotective activities in a murine neuroinflammatory model. METHODS: OM phytochemical analysis was done by HPLC/PDA/ESI-MSn. Oxidative stress was induced in vitro by hydrogen peroxide and cell viability was measured using WST-1 assay. Swiss albino mice were injected intraperitoneally with OM extract at a dose of 100 mg/kg for 12 days and with 250 µg/kg LPS daily starting from day 6 to induce neuroinflammation. Cognitive functions were assessed by novel object recognition and Y-maze behavioral tests. Hematoxylin and eosin staining was used to assess the degree of neurodegeneration in the brain. Reactive astrogliosis and inflammation were assessed by immunohistochemistry using GFAP and COX-2 antibodies, respectively. RESULTS: OM is rich in phenolics, with rosmarinic acid and its derivatives being major constituents. OM extract and rosmarinic acid significantly protected microglial cells against oxidative stress-induced cell death (p < 0.001). OM protected against the LPS-induced alteration of recognition and spatial memory in mice (p < 0.001) and (p < 0.05), respectively. Mice that received OM extract prior to the induction of neuroinflammation showed comparable histology to control brains, with no overt neurodegeneration. Furthermore, OM pre-treatment decreased the immunohistochemistry profiler score of GFAP from positive to low positive and COX-2 from low positive to negative in the brain tissue, compared to the LPS group. CONCLUSION: These findings highlight the potential preventive effects of OM phenolics against neuroinflammation and pave the way toward drug discovery and development for neurodegenerative disorders.

Origanum vulgare Essential Oil Modulates the AFB1-Induced Oxidative Damages, Nephropathy, and Altered Inflammatory Responses in Growing Rabbits.

The current study was performed to investigate the toxic effects of aflatoxin B1 (AFB1) through the evaluation of kidney function tests and histopathological examination of renal tissues, targeting the therapeutic role of Marjoram (Origanum vulgare essential oil-OEO) in improving health status. Forty-eight New Zealand Whites growing rabbits (four weeks old) weighing on average 660.5 ± 2.33 g were randomly and equally distributed into four groups, each of which had four replicas of three animals as the following: Control group (only basal diet), AFB1 group (0.3 mg AFB1/kg diet), OEO group (1 g OEO/kg diet) and co-exposed group (1 g OEO/kg + 0.3 mg AF/kg diet). Our study lasted eight weeks and was completed at 12 weeks of age. The results revealed that OEO decreased the toxic effects of AFB1 in rabbit kidneys by substantially reducing the cystatin C levels in the AFB1 group. Additionally, OEO decreased oxidative stress and lipid peroxidation levels in the co-exposed group. Moreover, OEO reduced DNA damage and inflammatory response in addition to the down-regulation of stress and inflammatory cytokines-encoding genes. Besides, OEO preserved the cytoarchitecture of rabbits' kidneys treated with AFB1. In conclusion, O. vulgare essential oil supplementation ameliorated the deleterious effects of AFB1 on the rabbits' kidneys by raising antioxidant levels, decreasing inflammation, and reversing oxidative DNA damage.

Evaluation of the Maturity and Gene Expression of Sheep Oocytes and Embryos Cultured in Media Supplemented with Marjoram (Origanum vulgare) Extract.

This study aimed to evaluate the effects of marjoram extract on oocyte maturation and gene expression in sheep oocytes and embryos. The first experiment studied the effect of the extract as an antioxidant to improve the in vitro maturation media used for sheep oocytes; the oocytes were matured in a TCM199 medium supplemented with 1 or 10 µg/mL of marjoram extract or the control, 0 µg, for 24 hr. Then, the maturation was estimated, and the gene expression was measured by using qPCR. The second experiment studied the effect of the extract on the development of sheep embryos produced in vitro; the fertilized oocytes were cultured in a SOF medium supplemented with 1 or 10 µg/mL of marjoram extract or the control, 0 µg, for 7 days. Then, the gene expression was measured using qPCR. The results showed that the marjoram extract did not improve nuclear maturation or the blastocyst rate. There was a significant increase in the level of GDF-9 gene expression in mature oocytes in the treatment groups. An increase in the expression of BCL-2 and EGR-1 genes was observed for the blastocysts in the 10 µg/mL group. We concluded that the marjoram extract did not improve nuclear maturation, but it did affect the expression of some genes in sheep oocytes and embryos.

A behavioral and molecular study; ameliorated anxiety-like behavior and cognitive dysfunction in a rat model of chronic unpredictable stress treated with oregano extract.

OBJECTIVE: Chronic stress is considered a severe risk factor leading to various disorders, including anxiety and cognitive decline. The present study aimed to investigate the effects of Origanum vulgare (oregano) extract on improving anxiety-like behavior and learning and memory defection caused by chronic unpredictable stress (CUS). METHOD: A 10-day CUS protocol was executed on male rats, and on day 10, their anxiety, learning, and memory status were evaluated. After that, in addition to the CUS, the rats were treated with the oregano extract for 2 weeks. Then, the expression of BDNF, TrkB, and TLR2/4 genes in the hippocampus and prefrontal cortex of the rats was evaluated. Also, the liver- and kidney-related serum parameters, including triglycerides, total cholesterol, HDL, LDL, creatinine, urea, serum glucose, alanine aminotransferase, and aspartate aminotransferase were assessed. Further, the extract's lethal effect and its impact on animals' body weight were investigated. RESULTS: Behavioral tests confirmed the anxiety-like behavior and learning-memory function impairment caused by CUS. In contrast, the administration of the extract could significantly alleviate the mental deficiencies and diminished anxiety-like behaviors. Molecular assessments showed that CUS could markedly decrease the BDNF and TrkB genes' expression levels while increasing that of TLR2 and TLR4. In contrast, in extract-treated animals, mRNA levels of BDNF and TrkB considerably increased, yet TLR2 and TLR4 mRNA levels reduced. Additionally, consumption of the extract caused weight gain, while having no lethality and detrimental effect on the liver and kidneys functions. CONCLUSIONS: These findings indicate the anxiolytic properties of the extract and its improving effect on cognitive dysfunction.

The comparison of the effect of Origanum vulgare L. extract and vitamin C on the gentamycin-induced nephrotoxicity in rats.

The gentamicin renal toxicity has been the focal point for much discussion. The objective of this study was to investigate the impacts of Origanum vulgare L. extract and vitamin C on gentamicin dose-dependent toxicity in rats' kidney. The present study was conducted on 60 male Wistar rats divided into ten experimental groups: control (untreated), G1, G2, G3 (100, 200, 300 mg/kg gentamicin), M1, M2 and M3 (500 mg/kg marjoram extract) + 100, 200 and 300 gentamicin, V1, V2 and V3 (Vitamin C 500 mg/kg) + 100, 200 and 300 of gentamicin. On the last day, the serum was separated from heart blood and the kidney tissues were extracted to measure the biochemical and oxidative stress parameters and histological changes. Kidney damage was confirmed as dose-dependent gentamicin by biochemical and pathological parameters. Urea, Blood Urea Nitrogen (BUN), and creatinine showed a significant increase in the G3 group compared to the control, M1, and V1 groups (p < 0.01). Catalase (CAT), Glutathione peroxidase (GPx), and Superoxide dismutase (SOD) showed a significant reduction in renal tissue in the G3 group compared to the other groups (p < 0.001). Malondialdehyde (MDA) concentration in the kidney tissue of the G3 group also showed a significant increase compared to other groups (p < 0.001). Furthermore, TNFα and IL-1 levels were the highest in the G3 group, and serum total antioxidant capacity (TAC) concentration had the lowest amount compared to other groups. Moreover, histopathological lesions of the kidney showed significant statistical differences among the groups that received gentamicin with the control and M1 group. Marjoram extract at the dose of 500 mg/kg had a desirable effect on controlling gentamicin damage in the kidneys compared with vitamin C. In particular, controlling gentamicin-induced oxidative stress and inflammation by the consumption of marjoram extract and vitamin C plays an important role in protecting the kidneys.

Multifunctional poly(vinyl alcohol) films using cellulose nanocrystals/oregano and cellulose nanocrystals/cinnamon Pickering emulsions: Effect of oil type and concentration.

Poly (vinyl alcohol) (PVA) films with high transparency, UV-barrier, antioxidant, and antimicrobial properties were prepared using oregano essential oil (OEO) and cinnamon essential oil (CEO) Pickering emulsions. The effect of Pickering emulsion type and concentration on the PVA film properties was studied. Cellulose nanocrystals (CNCs) were used as a natural stabilizer to prepare OEO and CEO Pickering emulsions. Both emulsions showed spherical droplets with diameters of 155-291 nm, zeta potential of -36.2 to -49.6 mV, minimum inhibition concentration of 6.25-12.5 µL/mL, and inhibition zone of 40-65 mm, depending on oil type. Morphology and FTIR analysis showed that OEO and CEO Pickering emulsions were compatible with the PVA matrix. The UV-transmittance of PVA films decreased from 77.3% to 30.4% and 2.0% without sacrificing the transparency after adding OEO and CEO Pickering emulsions, respectively. Antimicrobial results showed that E. coli was more sensitive to CEO, while S. aureus was sensitive to OEO Pickering emulsion. PVA/CEO film displayed higher properties than PVA/OEO film.

Co-delivery of imidazolium Zn(II)salen and Origanum Syriacum essential oil by shrimp chitosan nanoparticles for antimicrobial applications.

This study reports preparation and physicochemical characterization of natural antimicrobials (Origanum Syriacum essential oil (OSEO), shrimp chitosan nanoparticles (CSNPs)) and new imidazolium ionic liquid-supported Zn(II)Salen. These antimicrobials were separately or co-encapsulated by CSNPs to fabricate novel antimicrobial nanoplatforms "NPFs" (OSEO-loaded CSNPs (NPF-1), Zn(II)Salen-loaded CSNPs (NPF-2), and Zn(II)Salen@OSEO-loaded CSNPs (NPF-3)). The finding of loading, encapsulation, and antimicrobial release studies confirm the suitability of CSNPs for nanoencapsulation of Zn(II)Salen and OSEO. All NPFs can significantly suppress the growth of microbial species with performances dependent upon the microbial strain and nanoplatform concentration. The susceptibility of microbes toward new antimicrobials was as follows; Gram-positive bacteria > Gram-negative bacteria > fungi. The amazing physicochemical features of new nanoplatforms and their bioactive ingredients (Zn(II)Salen, OSEO, and CSNPs) signify the importance of our designs for developing a new generation of nanopharmaceuticals supported both natural products and biogenic ionic metal cofactors, targeting the multidrug resistant (MDR) pathogens.

Growth, water use, yield and quality parameters in oregano affected by reduced irrigation regimes.

BACKGROUND: Determining plant tolerance to water stress is necessary in irrigation scheduling, decisions concerning supplementary irrigation, planning and operation and, more importantly, the rational use of water resources. In the present study, effects of the irrigation regime on oregano growth, water use, yield and quality parameters were investigated using reduced irrigation regime treatments. RESULTS: Increased water stresses caused a decrease in all growth and yield parameters. Plant water use efficiency was not significantly different among the control, low and medium stress treatments, although it was the lowest for the extreme water stress treatment. Total essential oil yield was the only quality parameter demonstrating significant differences among treatments. Yield response factors were determined as 1.13, 1.12, 1.06 and 1.10 for total fresh, total dry, dry leaf and total essential oil yields, respectively. CONCLUSION: Yield response factors indicate that oregano plant is slightly sensitive to water stress. Although the yield parameters were affected by water stress, Origanum onites, under cultivation in recent years, might have developed a resistance mechanism for quality parameters because it has grown in very dry conditions under a natural environment for many years. However, the negative relationships between crop evapotranspiration versus essential oil, total phenolic and flavonoid contents and extract yield indicates that the quality parameters of oregano under low plant water consumption may be increased. © 2020 Society of Chemical Industry.

Natural Antibiotic Oregano in Hydroxyapatite-Coated Titanium Reduces Osteoclastic Bone Resorption for Orthopedic and Dental Applications.

Traditional infection prevention and treatment methods include synthetic antibiotics, which can cause severe adverse side effects. Carvacrol and thymol are biologically active monoterpenoid extractants from oregano leaves with antibiotic capabilities; however, little is known regarding their effects on bone tissue engineering. The objective of this work is to understand their effects on osteogenesis, specifically with osteoblast and osteoclast cells, from surface-modified Ti6Al4V with plasma sprayed hydroxyapatite (HA) coatings. This system is an alternative to cemented implants to aid in bone healing. Results reveal that full carvacrol release from the HA matrix is successful in aqueous environments and modulation of release kinetics can also be made using polycaprolactone (PCL) and polyethylene glycol (PEG) polymers. From HA-pressed disc samples in physiological pH, full carvacrol release is achieved in 10 days using PCL/PEG, about 95% release in 50 days using no polymer, and 60% in 50 days when using a PCL coating. Without polymer, full carvacrol release is achieved after 3 days from HA coatings in both physiological pH and acidic pH, mimicking the post-surgery environment. The release is assessed as a diffusion-based mechanism in phosphate-buffered saline but degradation-based mechanism in acetate buffer solution. Carvacrol and thymol show bacterial inhibition of Staphylococcus epidermidis and no cytotoxic effects on osteoblast proliferation in vitro. Carvacrol and thymol also induce a significant 7% reduction in osteoclast tartrate-resistant acid phosphatase (TRAP) activity, caused by poorly attached cellular morphologies, leading to an approximately 65% reduction in osteoclast resorption pit formation. Our goal is to demonstrate a natural medicinal system that can support bone healing while providing infection prevention and reducing costly revision surgeries for orthopedic and dental applications.

Dietary Origanum vulgare essential oil attenuates cypermethrin-induced biochemical changes, oxidative stress, histopathological alterations, apoptosis, and reduces DNA damage in Common carp (Cyprinus carpio).

The study was designed to evaluate the possible protective roles of dietary Origanum vulgare essential oil (OVEO) against cypermethrin (CP)-induced serum biochemical changes and oxidative stress of common carp (Cyprinus carpio). Moreover, histopathological alterations, apoptosis, cell proliferation, and DNA damage in the gills and hepatic tissues were also assessed. Briefly, fish were allotted into six groups with three triplicates whereas a group fed on basal diet and did not exposed to CP and served as control (CTR), two groups were fed on diets supplemented with two levels of OVEO (0.5 % and 1.0 %), a group exposed to sub-lethal concentration of CP (1/10 of 96 h-LC50 = 0.4134 µg/L), and two other groups exposed to the same concentration of CP and fed on diets supplemented with both levels of OVEO (CP + 0.5 % OVEO, and CP + 1.0 % OVEO), respectively, for 30 days. CP induced significant elevation of serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), urea, and creatinine levels indicating hepato-renal toxicity (P < 0.05). Besides, there was a significant decrease in serum catalase (CAT) and superoxide dismutase (SOD) activities (P < 0.05). Moreover, CP induced significant histopathologic alterations in gills, anterior kidneys, and hepatic tissues with activation of apoptosis (Caspase-3) and proliferating cell nuclear antigen (PCNA). Comet assay demonstrated significant DNA damage in gills and liver tissues of the CP-exposed group. Interestingly, a significant attenuation of serum ALT, AST, ALP, urea, creatinine, CAT, and SOD levels (P < 0.05) was noticed in CP-exposed fish and concurrently fed diets supplemented with either 0.5 % or 1.0 % OVEO. Moreover, histopathologic alterations and apoptosis were significantly reduced along with a concomitant significant decrease in DNA damage (P < 0.05) which indicated the mitigation of DNA damage. Conclusively, the study showed that OVEO is an effective counteractive treatment against CP-induced damage in exposed common carp and is recommended during the formulation of fish rations.

Genome sequencing of four culinary herbs reveals terpenoid genes underlying chemodiversity in the Nepetoideae.

Species within the mint family, Lamiaceae, are widely used for their culinary, cultural, and medicinal properties due to production of a wide variety of specialized metabolites, especially terpenoids. To further our understanding of genome diversity in the Lamiaceae and to provide a resource for mining biochemical pathways, we generated high-quality genome assemblies of four economically important culinary herbs, namely, sweet basil (Ocimum basilicum L.), sweet marjoram (Origanum majorana L.), oregano (Origanum vulgare L.), and rosemary (Rosmarinus officinalis L.), and characterized their terpenoid diversity through metabolite profiling and genomic analyses. A total 25 monoterpenes and 11 sesquiterpenes were identified in leaf tissue from the 4 species. Genes encoding enzymes responsible for the biosynthesis of precursors for mono- and sesqui-terpene synthases were identified in all four species. Across all 4 species, a total of 235 terpene synthases were identified, ranging from 27 in O. majorana to 137 in the tetraploid O. basilicum. This study provides valuable resources for further investigation of the genetic basis of chemodiversity in these important culinary herbs.

Potential synergistic antimicrobial efficiency of binary combinations of essential oils against Bacillus cereus and Paenibacillus amylolyticus-Part A.

The checkerboard method was used to study the potential interactions between eight essential oils (Basil, Cinnamon, Eucalyptus, Mandarin, Oregano, Peppermint, Tea tree, and Thyme) when used as antibacterial agents against Bacillus cereus LSPQ 2872 and Paenibacillus amylolyticus ATCC 9995. The minimum inhibitory concentration (MIC) of each essential oil (EO) and the fractional inhibitory concentration (FIC) index for the binary combinations of essential oils (EOs) were determined. According to FIC index values, some of the compound binary combinations showed an additive effect; however, Thyme/Tea tree and Cinnamon/Thyme EOs exhibited a synergistic effect against P. amylolyticus and B. cereus, respectively. Cinnamon/Thyme EOs mixture exhibited no interactive effect against P. amylolyticus, but a synergistic effect against B. cereus. The combination of Oregano/Thyme EOs displayed the best antibacterial activity and showed a synergistic effect against B. cereus and P. amylolyticus bacteria. The Oregano/Thyme EOs mixture has potential application in food preservation to reduce the contamination of B. cereus and P. amylolyticus.

In vitro bioaccessibility and activity of Greek oregano (Origanum vulgare L. ssp. hirtum (link) Ietswaart) compounds as affected by nitrogen fertilization.

BACKGROUND: Greek oregano is a culinary and medicinal herb native to the Mediterranean region; however nowadays it is cultivated in many regions all over the world. It is commonly used as a spice for flavoring food products and in various traditional medicine applications. This study investigated the effect of nitrogen fertilization at 0-150 kg N ha-1 on the potential bioaccessibility and activity of Greek oregano compounds. For this purpose the total phenolic content, rosmarinic acid content, and antioxidant activities of the raw material, as well as digested fractions, were determined. RESULTS: Nitrogen fertilization had a negative influence on the phenolic content and antioxidant activity of raw material; however, its effect on the potential bioaccessibility varied depending on the dose. The highest potential bioaccessibility and activity was determined for plants fertilized with 30 kg N ha-1 . For the gastric and intestinal phases of digestion, the potential bioaccessibility percentages were 39.5% and 29.6% for total phenolics, 53.1% and 11.2% for rosmarinic acid content, 45.2% and 44.4% for antiradical activity against 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 39.2% and 27.2% for antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 50.2% and 23.4% for reducing power, respectively. CONCLUSION: The results showed that nitrogen fertilization is an important factor determining the in vitro bioaccessibility of Greek oregano compounds. Furthermore, the nutraceutical potential of herbs, in term of the bioaccessibility of bioactive compounds, may be optimized during plant cultivation by applying an accurate nitrogen level. © 2020 Society of Chemical Industry.

Uptake of Cd, Pb, and Ni by Origanum syriacum produced in Lebanon.

Trace metals are found naturally in soil. However, the increase in industrial and agricultural polluting activities has increased trace metal contamination and raised high concerns in the public health sector. The study was conducted on Origanum syriacum, one of the most consumed herbs in the Middle East, and was divided into three parts. (1) Pot experiment: to study the effect of Cd, Pb, or Ni levels in soil on their uptake by O. syriacum. (2) Field samples: collected from major agricultural regions in Lebanon to analyze Cd, Pb, and Ni concentrations in soil and leaves. (3) Sale outlets samples: to measure the levels of Cd, Pb, and Ni in O. syriacum tissues in the market. Results showed that there was a positive correlation between levels of Cd, Pb, and Ni in soil and those in O. syriacum tissues. None of the field samples contained Pb or Ni that exceeded the maximum allowable limits (MAL). Three samples collected from heavily poultry-manured soil contained Cd higher than the MAL. Samples collected from sale outlets did not exceed the MAL for Ni but two exceeded the MAL for Cd and one for Pb. Trace metal contamination is not a major concern in O. syriacum produced in Lebanon. Only one mixture sample from a sale outlet was higher in Pb than the MAL and three samples from heavily manured fields exceeded the MAL for Cd.

Determination of phenolic compounds by MALDI-TOF and essential oil composition by GC-MS during three development stages of Origanum majorana L.

This study aimed to investigate the effect of the maturation process of sweet marjoram (Origanum majorana L.) on essential oil composition, the phenolic profile of ethanolic extract and their antioxidant capacities. The essential oil composition was studied at three stages of maturity by GC-MS. Thirty compounds were detected representing 100% of the total essential oil. p-Menth-1-en-4-ol was the major compound (37.15-76.94%) followed by cyclohexanol-3,3,5 trimethyl (5.41-15.99%) and α-terpineol (0.94-11.34%). During the maturation process, an accumulation of oxygenated monoterpenes was observed. The phenolic composition was studied using matrix-assisted laser desorption/ionization time of flight. The analysis showed the presence of short flavonoid monomers at all stages of maturation. The antioxidant capacity of ethanolic extracts and essential oils was evaluated using the DPPH assay, iron chelating power and reducing power assay. The highest phenolic content and antioxidant capacity were found at flowering stage. These findings on essential oil composition, phenolic profile and antioxidant capacity of O. majorana at three different stages of development provide more information on how these secondary metabolites are accumulated.

Effect of oregano (Origanum vulgare L. ssp. hirtum) and clove (Eugenia spp.) nanoemulsions on Zygosaccharomyces bailii survival in salad dressings.

This work aimed to evaluate the in vitro effect of encapsulated oregano and clove essential oils on oil-in-water nanoemulsions against Zygosaccharomyces bailii. The antifungal efficacy of these nanoemulsions and their sensory acceptance were tested in salad dressings. Both essential oils were effective inhibitors against the target yeast, with minimal inhibitory and fungicidal concentrations of 1.75 mg/mL. In the in vitro assay done with the nanoemulsions, no yeast growth was observed for any tested essential oil concentration. In the salad dressings, all the formulations were able to reduce Z. bailii growth compared to the control, and only those samples with 1.95 mg/g of essential oil were capable of inhibiting yeast development after 4 inoculation days. The sensory acceptance of the dressing containing the nanoemulsions was similar to the control dressing in appearance, consistency and colour terms. These results evidence the antifungal activity of oregano and clove nanoemulsions against Z. bailii.

Phenolic compounds, antioxidant capacity and antimutagenic activity in different growth stages of in vitro raised plants of Origanum vulgare L.

Efficient micropropagation procedure was developed for Origanum vulgare, a high-value culinary herb, and the phytochemicals, phenolic content, antioxidant and antimutagenic activity of leaf and stem, derived from different growing stages were analyzed. The agar solidified Murashige and Skoog (MS) medium supplemented with a combination of 6-benzylaminopurine and α-naphthaleneacetic acid was optimized as best shoot-multiplication-medium. Shoots were rooted best on 1/2 strength MS medium supplemented with 50 µM indole-3-butyric acid (IBA). The plantlets were successfully acclimatized ex vitro in a soil, sand and farmyard manure mixture (2:1:1 v/v/v) with 100% survival rate in greenhouse. The total anthocyanin and total phenolic content were observed significantly higher in leaves of in vitro-raised plants. However, total tannin, flavonoid and antioxidant activity remained higher in leaves of mother plant maintained under ployhouse condition. All the plant extracts have shown significant antimutagenic activity except in vitro-growing plants. A total of 13 polyphenolic compounds were detected in different extracts using high performance liquid chromatography. Among these, catechin was detected maximum in in vitro-growing cultures and chlorogenic acid in leaves of mother plant. These findings will help the farmers, medicinal plant growers, and industries for mass multiplication and effective extraction of phytochemicals from O. vulgare.

Phytochemical Profiles and Antibacterial Activities of Hydroalcoholic Extracts of Origanum vulgare and Hypericum perforatum and Carvacrol and Hypericin as a Promising Anti-Staphylococcus aureus.

OBJECTIVES: Staphylococcus aureus, a Gram-positive bacteria, is ranked second among the causes of hospital infections and is one of the three main causes of food poisoning. In recent times, the spread of antibiotic resistance in S. aureus has become very worrisome. Therefore, research for new effective drugs is important. The present study aims to investigate the phytochemical profiles and antibacterial effects of hydroalcoholic extracts of Origanum vulgare (Lamiaceae family) and Hypericum perforatum (Clusiaceae family) and their active compounds on S. aureus (ATCC 12600) in vitro. METHODS: The identification of phytochemical compounds in both plants was performed by Highperformance liquid chromatography (HPLC), headspace-solid-phase microextraction (HS-SPME) and Fourier-transform infrared spectroscopy (FTIR). To investigate microbial susceptibility, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and disc diffusion method (DAD) were used. Finally, the results of the study were compared with methicillin. RESULTS: Of the 42 combinations of O. vulgare, carvacrol (48%) and of the 38 combinations of H. perforatum, hypericin (46.2%) were the most abundant. The MIC, MBC and DAD of O. vulgare and H. perforatum, carvacrol, hypericin and methicillin were 625, 625, 312.5, 78.12 and 384 µg/mL, 10000, 10000, 2500, 2500 and 384 µg/mL, and 15.66 ± 4.49, 12.66 ± 0.47 and 22 ± 0.81 mm, respectively. CONCLUSION: Due to the significant effects of O. vulgare and H. perforatum and their active components against S. aureus, it is expected that in the future, hypericin, carvacrol and their derivatives can be used as effective antibacterial agents against S. aureus.