Therapeutic benefit of oregano oil in the acute idiosyncratic hepatotoxicity induced by carbon tetrachloride in rats: Adverse effects of high dose of oreganum.

Toxicity caused by carbon tetrachloride (CCl4) can lead to serious liver injury. The aim of the study is to investigate the protective effects of oregano oil (Origanum minutiflorum extract oil) against CCl4-induced liver injury. Two doses of oregano oil were used in the experiment: a low dose (LD; 20 mg/kg) and a high dose (HD; 60 mg/kg) during 2 weeks. CCl4 caused severe liver damage, nucleolus destruction in hepatocytes and cytogenetic changes in the nucleus. Indirectly, CCl4 causes decreased protein synthesis and significantly high creatinine and urea values. Hematological disorders have been recorded, such as decreased RBC and hemoglobin concentration, increased WBC and deformability of the erythrocyte membrane. Both doses of oregano oil had protective effects. Improved protein synthesis and high globulins level, creatinine and urea were found in both groups. Cytogenetic changes in the nucleus of hepatocytes were reduced. A high dose of oregano oil had maximal protective effects for RBC, but a very weak effect on hemoglobin synthesis. Also, WBC and lymphocyte values were low. Origanum stimulates protein synthesis and recovery of hepatocytes after liver injury, reduces the deformability of the erythrocyte membrane. High doses of oregano oil decreased WBC and lymphocytes which may lead to a weakening of the immune response. However, high doses are more effective against severe platelet aggregation than low doses, suggesting an effective treatment against thrombocytosis.

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.

The impact of Origanum Vulgare essential oil supplementation on sperm motility and subpopulation alterations in bulls, dogs, and rabbits.

Artificial insemination is a widely adopted method in livestock production for various reasons such as health security and genetic improvement. Although sperm motility is of paramount importance in this technique as it directly influences the sperm's ability to fertilize the oocyte. In previous research on human sperm, we observed that in vitro supplementation with Origanum Vulgare essential oil significantly improved sperm motility and antioxidant activities, all without negatively affecting the integrity of their DNA. Based on these promising results, we considered it crucial to explore the potential effects of supplementation with this essential oil on sperm of other species. In this study, we studied the effects of oregano essential oil supplementation on sperm motility of (bulls = 15) (dogs = 15) and (rabbits = 9) and the changes that in vitro incubation with this oil could induce on sub-motile sperm populations of different species. The results of the study showed that in vitro oregano essential oil supplementation had a significant impact on sperm motility in the three species studied. This improvement in sperm motility was accompanied by an increase in the proportion of subpopulations with high velocity and progressivity: an increase of (2.16%, 10% and 4.84%) for subpopulation 1, (6.50%, 5.5% and 3.17%) for subpopulation 4 in bulls, dogs and rabbits respectively. While the subpopulations representing low motile and non-progressive sperm have decreased. These results suggest that the use of oregano essential oil can be a beneficial approach to improve sperm motility in different species, which can have important implications for the success of artificial insemination.

Essential oil and plant extract of oregano as agents influencing the virulence factors of Candida albicans.

Candida albicans, a polymorphic yeast, is a physiological component of the human and animal commensal microbiome. It is an etiological factor of candidiasis, which is treated by azole antifungals. Growing resistance to azoles is a reason to look for other alternative treatment options. The pharmacotherapeutic use of plant extracts and essential oils has become increasingly important. In our experiment, C. albicans showed susceptibility to four observed plant extracts and essential oils from peppermint ( Mentha piperita), thyme ( Thymus vulgaris), sage ( Salvia officinalis), and oregano ( Origanum vulgare). Oregano plant extract and essential oil showed the highest antifungal activity, at MIC values of 4.9 mg/mL and 0.4 mg/mL respectively. Therefore, it was subjected to further research on the influence of virulence factors - biofilm formation, extracellular phospholipase production and germ tube formation. Oregano plant extract and essential oil showed an inhibitory effect on the observed C. albicans virulence factors at relatively low concentrations. The extract inhibited the adherence of cells at MIC 12.5 mg/mL and essential oil at MIC 0.25 mg/mL. Degradation of the formed biofilm was detected at MIC 14.1 mg/mL for plant extract and at MIC 0.4 mg/mL for essential oil. Extracellular phospholipase production was most effectively inhibited by the essential oil. In particular, the number of isolates with intensive extracellular phospholipase production decreased significantly. Of the 12 isolates intensively producing extracellular phospholipase, only 1 isolate (4.5%) retained intense production. Essential oil caused up to a 100 % reduction in germ tubes formation and plant extract reduced their formation depending on the concentration as follows: 2.6% (0.8 mg/mL), 21.2 % (6.25 mg/mL), and 64.5 % (12.5 mg/mL) compared to the control.

Antibacterial Properties of Bacterial Endophytes Isolated from the Medicinal Plant Origanum heracleoticum L.

BACKGROUND: Bacterial endophytic communities associated with medicinal plants synthesize a plethora of bioactive compounds with biological activities. Their easy isolation and growth procedures make bacterial endophytes an untapped source of novel drugs, which might help to face the problem of antimicrobial resistance. This study investigates the antagonistic potential of endophytic bacteria isolated from different compartments of the medicinal plant O. heracleoticum against human opportunistic pathogens. METHODS: A panel of endophytes was employed in cross-streaking tests against multidrug-resistant human pathogens, followed by high-resolution chemical profiling using headspace-gas chromatography/mass spectrometry. RESULTS: Endophytic bacteria exhibited the ability to antagonize the growth of opportunistic pathogens belonging to the Burkholderia cepacia complex (Bcc). The different inhibition patterns observed were related to their taxonomic attribution at the genus level; most active strains belong to the Gram-positive genera Bacillus, Arthrobacter, and Pseudarthrobacter. Bcc strains of clinical origin were more sensitive than environmental strains. Cross-streaking tests against other 36 human multidrug-resistant pathogens revealed the highest antimicrobial activity towards the Coagulase-negative staphylococci and Klebsiella pneumoniae strains. Interestingly, strains of human origin were the most inhibited, in both groups. Concerning the production of volatile organic compounds (VOCs), the strain Arthrobacter sp. OHL24 was the best producer of such compounds, while two Priestia strains were good ketones producers and so could be considered for further biotechnological applications. CONCLUSIONS: Overall, this study highlights the diverse antagonistic activities of O. heracleoticum-associated endophytes against both Bcc and multidrug-resistant (MDR) human pathogens. These findings hold important implications for investigating bacterial endophytes of medicinal plants as new sources of antimicrobial compounds.

Enhancing effect of oregano essential oil and Bacillus subtilis on broiler immune function, intestinal morphology and growth performance.

The present study evaluated the effect of two categories of feed additives on chicken performance through immunological and intestinal histo-morphometric measurements. A total of 150 one-day-old male broiler chicks (Cobb) were randomly assigned to three groups. Group I received a non-supplemented basal diet. While groups II and III were treated with a basal diet supplemented with oregano essential oil (OEO) and Bacillus subtilis, respectively, in water for 28 days. Blood samples were taken at 6, 18 and 28 days for hematological analysis, phagocytosis, lymphocyte proliferation and measuring antibody responses. Additionally, growth performance indices were recorded weekly. The results showed that groups supplemented with OEO and B. subtilis improved growth performance expressed by a significant increase in weight gain (P < 0.05), with a significant reduction (P < 0.05) in feed conversion ratio (FCR). Hematological findings indicated a significant increase in blood parameters as well as a significant increase in phagocytic % & phagocytic index at all time points with a greater probiotic effect. On the other hand, OEO produced a significant increase in lymphocyte proliferation at 18 & 28 days. Humoral immunity revealed a significant increase in serum antibody titer phytobiotic & probiotic-fed groups at time points of 18 & 28 days with a superior phytobiotic effect. The histological examination showed a significant increase in villi length, villi width, crypt depth & V/C ratio. In conclusion, these results indicated positive effects of B. subtilis & OEO on both growth and immunity and could be considered effective alternatives to the antibiotic.

Calcium Channel Inhibitory Effect of Marjoram (Origanum majorana L.): Its Medicinal Use in Diarrhea and Gut Hyperactivity.

BACKGROUND: The leaves of Origanum majorana (O. majorana) are traditionally renowned for treating diarrhea and gut spasms. This study was therefore planned to evaluate its methanolic extract. METHODS: Gas chromatography-mass spectrometry (GC-MS) was used to identify the phytochemicals, and Swiss albino mice were used for an in vivo antidiarrheal assay. Isolated rat ileum was used as an ex vivo assay model to study the possible antispasmodic effect and its mechanism(s). RESULTS: The GC-MS analysis of O. majorana detected the presence of 21 compounds, of which alpha-terpineol was a major constituent. In the antidiarrheal experiment, O. majorana showed a substantial inhibitory effect on diarrheal episodes in mice at an oral dosage of 200 mg/kg, resulting in 40% protection. Furthermore, an oral dosage of 400 mg/kg provided even greater protection, with 80% effectiveness. Similarly, loperamide showed 100% protection at oral doses of 10 mg/kg. O. majorana caused complete inhibition of carbachol (CCh, 1 µM) and high K+ (80 mM)-evoked spasms in isolated ileal tissues by expressing significantly higher potency (p < 0.05) against high K+ compared to CCh, similar to verapamil, a Ca++ antagonist. The verapamil-like predominant Ca++ ion inhibitory action of O. majorana was further confirmed in the ileal tissues that were made Ca++-free by incubating the tissues in a physiological salt solution having ethylenediaminetetraacetic acid (EDTA) as a chelating agent. The preincubation of O. majorana at increasing concentrations (0.3 and 1 mg/mL) shifted towards the right of the CaCl2-mediated concentration-response curves (CRCs) with suppression of the maximum contraction. Similarly, verapamil also caused non-specific suppression of Ca++ CRCs towards the right, as expected. CONCLUSIONS: Thus, this study conducted an analysis to determine the chemical constituents of the leaf extract of O. majorana and provided a detailed mechanistic basis for the medicinal use of O. majorana in hyperactive gut motility disorders.

Synergistic effects of thyme and oregano essential oil combinations for enhanced functional properties of sericin/pectin film.

This work aimed to assess the synergistic antibacterial effects of thyme and oregano essential oils in various ratios (thyme:oregano; 10:0, 8:2, 6:4, 4:6, 2:8, 10:0). We hypothesized that the synergistic combination of thyme and oregano essential oils can be effectively incorporated into sericin/pectin film to enhance its functional properties. Among the combinations tested, the mixture of thyme/oregano essential oil (TOE) at an 8:2 ratio exhibited the most potent synergistic activity against P. aeruginosa and S. aureus, with fractional inhibitory concentration index (FICindex) of 0.9. In this combination, thymol constituting 51.83 % of TOE (8:2), was the predominant component. TOE at an 8:2 ratio was selected to incorporate into sericin/pectin film. Different concentrations of TOE (0.8 %, 1.2 % and 1.6 %) were applied to evaluate their impact on film properties compared to a film without essential oil (control). It was found that increasing TOE concentration (control; 0 %) to 1.6 % reduced film moisture content (from 21.53 % to 16.91 %), decreased yellowness (from 18.24 to 15.92), diminished gloss (from 63.79 to 11.18), lowered swelling index (from 1.24 to 0.98), and reduced tensile strength (from 9.70 to 4.14 MPa). However, the addition of TOE showed higher film total phenolic content (8.59-31.53 mg gallic acid/g dry sample) and increased antioxidant activity (0.99-3.68 µmol Trolox /g dry sample). Moreover, the film with 1.2 % and 1.6 % of thyme/oregano essential oil exhibited inhibitory effects against all tested bacteria. Therefore, the thyme/oregano essential oil combination can provide the desirable physicochemical properties of the sericin/pectin film, as well as its antibacterial and antioxidant activities, making it a promising alternative for food packaging material applications.

Exploring the phytoconstituents, antimicrobial potency, and cytotoxic effects of essential oil from Origanum punonense from Palestine.

BACKGROUND: Origanum punonense Danin is one of the old traditional medicinal plants Bedouins utilize in the Dead Sea region to treat a variety of illnesses, those caused by infections. The current study aimed to identify the phytochemical components of O. punonense essential oil (EO) and determine its antiproliferative and antimicrobial effects. METHODS: Gas chromatography and mass spectrometry were employed to detect the phytochemical constituents of O. punonense EO. Broth microdilution assay was utilized to determine the antimicrobial effects against various microbial species, including those causing diabetic foot infections. RESULTS: This study revealed that O. punonense EO contains 44 phytochemical compounds, of which 41 compounds were detectable and amounted to 99.78% of the total oil. The main chemical components of the oil were carvacrol (57.4%), p-cymene (6.66%), carvone (5.35%), pinene (4.9%), and terpinene (2.96%). The antiproliferative activity of different concentrations of O. punonense EO was noted in all of the investigated cell lines, with the best activity at the concentration of 500 µg/mL. The greatest antibacterial activity was against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Proteus vulgaris, with MIC values of 1.56 µL/mL. In addition, and the O. punonense EO showed strong antifungal activity against Candida albicans with a MIC value of 0.8 µL/mL. In addition, the O. punonense EO showed potent antibacterial activity against all MRSA samples obtained from the diabetic foot with a MIC value of 3.13 µL/mL. The O. punonense EO demonstrated potent activity against Carbapenem-resistant Enterobacterales, Citrobacter freundii, and K. pneumoniae, with MICs value of 6.25 µL/mL. CONCLUSION: The potent antiproliferative and broad antimicrobial activity of O. punonense EO makes it an effective strategy for treating infections, especially in immunocompromised patients with chronic comorbidities such as cancer and diabetes mellitus.

Oregano (Origanum vulgare) Essential Oil and Its Constituents Prevent Rat Kidney Tissue Injury and Inflammation Induced by a High Dose of L-Arginine.

This study aimed to evaluate the protective action of oregano (Origanum vulgare) essential oil and its monoterpene constituents (thymol and carvacrol) in L-arginine-induced kidney damage by studying inflammatory and tissue damage parameters. The determination of biochemical markers that reflect kidney function, i.e., serum levels of urea and creatinine, tissue levels of neutrophil-gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1), as well as a panel of oxidative-stress-related and inflammatory biomarkers, was performed. Furthermore, histopathological and immunohistochemical analyses of kidneys obtained from different experimental groups were conducted. Pre-treatment with the investigated compounds prevented an L-arginine-induced increase in serum and tissue kidney damage markers and, additionally, decreased the levels of inflammation-related parameters (TNF-α and nitric oxide concentrations and myeloperoxidase activity). Micromorphological kidney tissue changes correlate with the alterations observed in the biochemical parameters, as well as the expression of CD95 in tubule cells and CD68 in inflammatory infiltrate cells. The present results revealed that oregano essential oil, thymol, and carvacrol exert nephroprotective activity, which could be, to a great extent, associated with their anti-inflammatory, antiradical scavenging, and antiapoptotic action and, above all, due to their ability to lessen the disturbances arising from acute pancreatic damage. Further in-depth studies are needed in order to provide more detailed explanations of the observed activities.

Efficacy and safety of geranium-oregano-thymol formulations to control of dog tick Rhipicephalus sanguineus sensu lato under laboratory and field conditions.

The present study evaluated, in laboratory and field, the efficacy and safety of formulations of Pelargonium graveolens (geranium - G), Origanum majorana (oregano - O) commercial essential oils (EO) and thymol (T) to control of Rhipicephalus sanguineus sensu lato. In the laboratory, three formulas (A: 2% tween 80%, B: powder and C: nanoemulsion) by a mixture of these components (GOT) were prepared and evaluated, and the best one was used to assess its safety and field application against R. sanguineus s. l. on naturally infested dogs. Besides the major compounds of the EO used were identified. The results of the lab study showed that formula A (2.5 g of each G + O + T + 2% tween 80 to complete 100 mL) was significantly more effective than the other two formulas tested and exhibited highly effective adulticidal, larvicidal, and ovicidal activity against R. sanguineus s.l. Significant LC50 and LC90 values of GOT were evaluated (13.4 and 21.5 mg/mL, respectively) for the adulticidal activity, (2.81 and 4.46 mg/mL, respectively) for ovicidal activity and (2.44 and 4.45 mg/mL, respectively) for larvicidal activity. The safety of formula A has been proven by the absence of its cytotoxicity on a cell line of human epidermoid carcinoma. Citronella and carvacrol were the major compounds identified in the commercial essential oils of P. graveolens and O. majorana, respectively. Formula A was used in a field control trial for almost 8 months, during the tick infestation season (April to November, 2022). Fourteen naturally infested dogs were divided into two groups, each with seven dogs. One group received formula A spraying five times during an experiment that continued for 8 months, while the other group received treatment with commercially available malathion acaricide. The animals were sprayed on five occasions throughout the experiment (April, June, July, August, and September). The results showed a substantial percentage of effectiveness after the first application of formula A with a 99.3% reduction in tick count at day 28 post-application (PA). In the case of severe infestation 60 days after the first application of formula A (more than 180 ticks per dog), the second application was done, achieving an efficacy of 54.9% at day 3 PA, so an emergency spray was done at day 5 PA to combat the rest of the tick infestation, achieving efficacy of 99% after 3 days. Consequently, a regular spray (third, fourth, and fifth application) was done every 35 days. This regular spray revealed 100% effectiveness at 14 days PA. Biochemical parameters of treated dogs were evaluated to confirm the safety of formula A. Creatinine, ALT, and albumin of the dogs treated with formula A were within the normal range of dogs, while urea and AST were higher than the normal range. In conclusion, formula A can safely treat R. sanguineus s.l. infestations in dogs with regular application every 5 weeks.

Chemical Composition, and Antioxidant and Antimicrobial Activity of Oregano Essential Oil.

Antimicrobial resistance (AMR) is a global public health problem, and the rapid rise in AMR is attributed to the inappropriate and/or overuse of antibiotics. Therefore, alternative antimicrobial agents, including those of natural origin, are being sought for the development of new drugs. The purpose of our study was to analyze the chemical composition, and antimicrobial and antioxidant activities of four oregano essential oils (OEOs) from Poland, Europe, Turkey and the USA. The antimicrobial activity (AMA) was evaluated using 23 strains, including Gram-positive bacteria, Gram-negative bacteria and Candida species. The antioxidant activity (AA) of essential oils (EOs) was determined by the DPPH method. The main component of the EOs tested was carvacrol (76.64-85.70%). The highest amount of this compound was determined in the Polish OEO. The OEOs we tested showed antimicrobial resistance, which was especially strong against fungi (MIC = 0.06-0.25 mg/mL-1). These products also showed high AA (71.42-80.44%). OEOs high in carvacrol should be the subject of further research as potential antimicrobial and antioxidant agents.

Growth performance, blood biochemistry, and mRNA expression of hepatic heat shock proteins of heat-stressed broilers in response to rosemary and oregano extracts.

The growing interest in countering the adverse effects of heat stress in poultry using phytogenic feed additives has garnered considerable attention in recent times, this research sought to examine the impact of rosemary leaves extract (RLE) and oregano leaves extract (OLE) on the growth performance, physiological responses, and hepatic mRNA expression of heat shock proteins in broiler chickens exposed to heat stress. A total of 150 male Indian River chicks, aged one day, were randomly allocated into five equally sized groups, each consisting of six replicates. The initial group was designated as the control and was provided with the basal diet. The second and third groups (R1 and R2) were administered the basal diet enriched with 50 and 100 mg/kg of rosemary leaves extract (RLE), respectively. The fourth and fifth groups (O1 and O2) were fed the basal diet supplemented with 50 and 100 mg/kg of oregano leaves extract (OLE), respectively. These chicks were reared in a controlled environmental chamber maintained at a temperature of 32±2 °C and relative humidity of 50 ± 5 %. Ferruginol was the leading component in RLE, whereas thymol was the prevalent constituent in OLE. RLE and OLE both have high DPPH• and ABTS•+ antioxidant potential. Among the experimental groups, the fourth group (O1) showed the heaviest live body weight and the lowest feed conversion ratio, indicating improved growth performance. There was a significant reduction in plasma total lipids and LDL-cholesterol levels within the R2 and O2 groups, respectively. Enhanced total antioxidant capacity and an improvement in the T3 hormone were observed in the R1 and R2 groups. In the second and fourth groups, the mRNA expression of hsp70 and 90A were both found to be significantly downregulated, respectively. In conclusion, the addition of 50 mg/kg of oregano leaves extract (OLE) to the diets of heat-stressed broilers resulted in improved hepatic heat shock proteins, along with certain physiological responses, ultimately contributing to enhanced growth performance.

Supercritical CO2 extraction of naringenin from Mexican oregano (Lippia graveolens): its antioxidant capacity under simulated gastrointestinal digestion.

A supercritical CO2 method was optimized to recover naringenin-rich extract from Mexican oregano (Lippia graveolens), a flavanone with high antioxidant and anti-inflammatory activity. The effect of the extraction parameters like pressure, temperature, and co-solvent on naringenin concentration was evaluated. We used response surface methodology to optimize the naringenin extraction from oregano; the chemical composition by UPLC-MS of the optimized extract and the effect of simulated gastrointestinal digestion on its antioxidant capacity and total phenolic content were also evaluated. The optimum conditions were 58.4 °C and 12.46% co-solvent (ethanol), with a pressure of 166 bar, obtaining a naringenin content of 46.59 mg/g extract. Also, supercritical optimized extracts yielded high quantities of cirsimaritin, quercetin, phloridzin, apigenin, and luteolin. The results indicated that the naringenin-rich extract obtained at optimized conditions had higher total phenolic content, antioxidant capacity by TEAC and ORAC, and flavonoid content, compared with the methanolic extract, and the simulated gastrointestinal digestion reduced all these values.

Investigation of physicochemical and biological properties of bacterial cellulose & zein-reinforced edible nanocomposites based on flaxseed mucilage containing Origanum vulgare L. essential oil.

In the present study, the effect of zein and different amounts of bacterial cellulose (BC; 1, 2 and 3 wt%) on the physical, mechanical and barrier properties of flaxseed mucilage/carboxymethyl cellulose (FM/CMC) composite was investigated. The appearance of the absorption band at 1320cm-1 in the ATR-FTIR spectra of nanocomposites indicated the successful introduction of zein into their structure. The characteristic peak at 2θ of 9° belonging to zein disappeared in XRD patterns of the prepared composites suggesting the successful coating of zein via hydrogen bonding interactions. SEM images proved the formation of semi-spherical zein microparticles in the FM/CMC matrix. TGA plots ascertained the addition of zein and nanocellulose caused a significant increase in the thermal stability of FM/CMC film, although zein showed a greater effect. The presence of zein and nanocellulose increased the mechanical strength of nanocomposites. The WVP of FM/CMC decreased after the incorporation of zein and nanocellulose, which created a tortuous path for the diffusion of water molecules. The zein particles exhibited a greater influence on improving the mechanical and barrier properties compared to nanocellulose. FM/CMC-Z film exhibited the highest mechanical strength (49.07 ± 5.89 MPa) and the lowest WVP (1.179 ± 0.076). The composites containing oregano essential oil (EO) showed higher than 60 % antibacterial properties. The bactericidal efficiency of FM/CMC/Z-EO and FM/CMC/Z-EO/BC1 nanocomposites decreased about 10% compared to FM/CMC/EO and FM/CMC-Z/BC1. This evidenced the successful encapsulation of EO molecules in zein particles. According to the in vitro release study, entrapment of EO into zein particles could delay the release and provide the extended antimicrobial effect.

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.

Effects of thermal and nonthermal treatments on microorganisms, pyrrolizidine alkaloids and volatile compounds in oregano (Origanum vulgare L.).

Effects of steam sterilization, gamma-irradiation, UV-irradiation and ozonation on microbial inactivation, pyrrolizidine alkaloid degradation and volatile compound profile in oregano were investigated. Steam sterilization and gamma-irradiation were the most effective treatments in inactivating microorganisms. These treatments resulted in 0.87-2.15 log reductions in total aerobic mesophilic bacteria counts and reduced yeast-mold and Enterobacteriaceae counts below the detectable level. Steam sterilization caused increased levels of pyrrolizidine alkaloids (PAs) and decreased levels of their N-oxide forms (PANOs) demonstrating a simultaneous conversion of PANOs into the corresponding PAs. Ozone treatment caused significant decreases in the levels of individual and total PAs/PANOs. After ozone treatment, decreases of 54.4, 53.9, 61.6 and 61.4% were observed in the levels of europine-N-oxide, europine, lasiocarpine-N-oxide and lasiocarpine, respectively. Steam sterilization, UV-irradiation and ozone treatments significantly altered the composition of the volatile compounds of oregano as evidenced by decreased levels of major components and the formation of some new compounds.

In ovo delivery of oregano essential oil activated xenobiotic detoxification and lipid metabolism at hatch in broiler chickens.

In ovo interventions are used to improve embryonic development and robustness of chicks. The objective of this study was to identify the optimal dose for in ovo delivery of oregano essential oil (OEO), and to investigate metabolic impacts. Broiler chickens Ross 308 fertile eggs were injected with 7 levels of OEO (0, 5, 10, 20, 30, 40, and 50 µL) into the amniotic fluid at embryonic d 17.5 (E17.5) (n = 48). Chick quality was measured by navel score (P < 0.05) and/or hatchability rates (P < 0.01) were significantly decreased at doses at or above 10 or 20 µL/egg, respectively, indicating potential toxicity. However, no effects were observed at the 5 µL/egg, suggesting that compensatory mechanisms were effective to maintain homeostasis in the developing embryo. To pursue a better understanding of these mechanisms, transcriptomic analyses of the jejunum were performed comparing the control injected with saline and the group injected with 5 µL of OEO. The transcriptomic analyses identified that 167 genes were upregulated and 90 were downregulated in the 5 µL OEO compared to the control group injected with saline (P < 0.01). Functional analyses of the differentially expressed genes (DEG) showed that metabolic pathways related to the epoxygenase cytochrome P450 pathway associated with xenobiotic catabolic processes were significantly upregulated (P < 0.05). In addition, long-chain fatty acid metabolism associated with ATP binding transporters was also upregulated in the OEO treated group (P < 0.05). The results indicated that low doses of OEO in ovo have the potential to increase lipid metabolism in late stages (E17.5) of embryonic development. In conclusion, in ovo delivery of 5 µL OEO did not show any negative impact on hatchability and chick quality. OEO elevated expression of key enzymes and receptors involved in detoxification pathways and lipid metabolism in the jejunum of hatchling broiler chicks.

Preparation and characterization of active films based on oregano essential oil microcapsules/soybean protein isolate/sodium carboxymethyl cellulose.

This study aimed to prepare oregano essential oil microcapsules (EOMs) by the active coalescence method using gelatin and sodium alginate as wall materials and oregano essential oil (OEO) as the core material. EOMs were added to the soybean protein isolate (SPI)/sodium carboxymethyl cellulose (CMC) matrix to prepare SPI-CMC-EOM active films, and the physical and chemical features of the active films and EOMs were characterized. The results showed that the microencapsulated OEO could protect its active ingredients. Scanning electron microscopy results showed that EOMs were highly compatible with the film matrix. The solubility of active films decreased upon adding EOMs, and their ultraviolet resistance and thermal stability also improved. When the added amount of EOMs was 5 %, the active films had the best mechanical properties and the lowest water vapor permeability. The active films prepared under this condition had excellent comprehensive performance. Also, adding EOMs considerably enhanced the antioxidant of the active films and endowed them with antibacterial properties. The application of the SPI-CMC-EOM films to A. bisporus effectively delayed senescence and maintained the freshness of the postharvest A. bisporus. This study provided a theoretical foundation for the incorporation of EOMs into active films based on biological materials.

Cytotoxic Effect Induced by Sicilian Oregano Essential Oil in Human Breast Cancer Cells.

Origanum vulgare L. is an aromatic plant that exerts antibacterial, antioxidant, anti-inflammatory, and antitumor activities, mainly due to its essential oil (EO) content. In this study, we investigated the possible mechanism underlying the in vitro antitumor activity of EO extracted by hydrodistillation of dried flowers and leaves of Origanum vulgare L. grown in Sicily (Italy) in MDA-MB-231 and MCF-7 breast cancer cell lines. Gas chromatography-mass spectrometry analysis of Oregano essential oil (OEO) composition highlighted the presence of twenty-six major phytocompounds, such as p-cymene, γ-terpinene, and thymoquinone p-acetanisole. OEO possesses strong antioxidant capacity, as demonstrated by the DPPH test. Our studies provided evidence that OEO reduces the viability of both MCF-7 and MDA-MB-231 cells. The cytotoxic effect of OEO on breast cancer cells was partially counteracted by the addition of z-VAD-fmk, a general caspase inhibitor. Caspases and mitochondrial dysfunction appeared to be involved in the OEO-induced death mechanism. Western blotting analysis showed that OEO-induced activation of pro-caspases-9 and -3 and fragmentation of PARP decreased the levels of Bcl-2 and Bcl-xL while increasing those of Bax and VDAC. In addition, fluorescence microscopy and cytofluorimetric analysis showed that OEO induces a loss of mitochondrial membrane potential in both cell lines. Furthermore, we tested the effects of p-cymene, γ-terpinene, thymoquinone, and p-acetanisole, which are the main components of OEO. Our findings highlighted that the effect of OEO on MDA-MB-231 and MCF-7 cells appears to be mainly due to the combination of different constituents of OEO, providing evidence of the potential use of OEO for breast cancer treatment.