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Watering level effect on Thymus hyemalis Lange essential oil yield and composition.

Thymus hyemalis Lange (chemotype thymol) was cultivated as an experimental crop under different watering level conditions in order to achieve 80, 60, 40, and 20% of the local potential evapotranspiration (Eto). Two harvesting periods were considered, winter and spring. As a consequence of the great variability among plants, essential oil yield percentages did not show statistically significant differences among seasons and different levels of water supply. Capillary GC-MS analysis of the essential oils permitted the detection of 84 volatile components. Among them, 54 are described for the first time as volatile constituents of the chromatographic profile of this Thymus species. Winter harvesting showed high concentrations in thymol percentage (25.92 +/- 4.39), the 40% Eto watering level being the best with respect to obtaining the optimum quality of this essential oil. However, in spring this thyme species needs a greater water supply (80% Eto) to achieve the same amount of thymol in the essential oil (29.20 +/- 2.83). From this, it was concluded that winter harvesting could be used for the extraction of the essential oil, with a low level of water supply, whereas spring harvesting could be employed for collection of leaves as a food condiment.

Gas Chromatography-Mass Spectrometry↗

Antioxidant properties of aqueous extracts from selected lamiaceae species grown in Turkey.

Water-soluble extracts from black thyme (Thymbra spicata L.), savory (Satureja cuneifolia Ten.), Spanish oregano (Coridothymus capitatus (L.) Reichb. f.), sweet marjoram (Majorana hortensis Moench), Syrian oregano (Origanum syriacum L.), Toka oregano (Origanum minutiflorum O. Schwarz et P. H. Davis), and Turkish oregano (Origanum onites L.) were screened for antioxidant properties in a battery of six in vitro assays. Total phenol content and qualitative-quantitative compositional analyses were also carried out. The extracts demonstrated varying degrees of efficacy in each screen. The savory extract was the most effective at reducing iron(III), scavenging 1,1-diphenyl-2-picrylhydrazyl radicals, inhibiting ascorbate-iron(III)-catalyzed hydroxyl radical-mediated brain phospholipid peroxidation, and site-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. The Syrian oregano extract was the most effective chelator of iron(II), while Spanish and Turkish oregano extracts were the most effective inhibitors of nonsite-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. All the extracts contained Folin-Ciocalteu reagent-reactive substances, which was confirmed by the presence of polar phenolic analytes (i.e., hydroxybenzoates, hydroxycinnamates, and flavonoids).

Antioxidants↗

Encapsulation of essential oils in zein nanospherical particles.

Three essential oils, oregano, red thyme, and cassia (100% pure oil), were encapsulated by phase separation into zein nanospheres. Topographical images indicated that the powders were made up of irregularly shaped particles ( approximately 50 mum) containing close-packed nanospheres. Approximately 31% of the oregano encapsulated particles had mean diameters greater than 100 nm compared to 19% for the zein alone particles. In vitro digestion of zein particles with pepsin at a concentration ratio of 10:1 was complete after 52 h in phosphate-citrate buffer, pH 3.5, at 37 degrees C by spectroscopic analysis. Nonenzymatic, aqueous in vitro release of essential oils from encapsulated zein particles was carried out in phosphate buffered saline at pH 7.4 and 37 degrees C. Release occurred at varying rates over 20 h probably from different locations within the closely packed nanospheres of different sizes. Gel electrophoresis SDS-PAGE of zein incubated with freeze-dried swine manure solids at 37 degrees C indicated that preformed microbial enzymes capable of digesting zein within minutes were present in the manure. Except for differences in size of nanospheres, no structural differences were resolved by several microscopic methods, suggesting that the oil and proteins phases were blended during phase separation.

Animals↗

Multivariate correlation between color and mineral composition of honeys and by their botanical origin.

The mineral content and color characteristics of 77 honey samples were analyzed. Eighteen minerals were quantified for each honey. Multiple linear regression (MLR) was used to establish equations relating the colorimetric CIELAB coordinates to the mineral data. The results obtained shown that lightness (L) was significantly correlated with S, Ca, Fe, As, Pb, and Cd for the dark honey types (avocado, heather, chestnut, and honeydew). For the light and brown honey types (citrus, rosemary, lavender, eucalyptus, and thyme), C(ab) and b showed the lower correlation with the mineral content of the honeys; their regression functions involve a few independent variables (Mg and Al for b and only Al for C(ab)). Furthermore, by means of application of linear discriminant analysis to the mineral content, it was possible to obtain a model that classifies the honeys by their lightness. The prediction ability of the built model, determined with the test set method, was 85%.

Analysis of Variance↗

The use of carbohydrate profiles and chemometrics in the characterization of natural honeys of identical geographical origin.

A study of the real possibilities of carbohydrate profiles and chemometrics to characterize the botanical origin of honey from a single geographical area, the Province of Soria (Spain), is presented. To this end, 14 carbohydrates were quantified using high-performance liquid chromatography (HPLC) with pulsed amperometric detection (PAD) in 77 natural honeys, the botanical origins of which were ling, spike lavender, French lavender, thyme, forest, and multifloral. Principal component analysis has been employed as a first approach to characterize the honey samples analyzed, showing similarities between spike lavender and multifloral honeys. The best discrimination among groups is obtained when four canonical discriminant analyses were carried out sequentially, origin by origin, achieving an overall percentage of success of 90% following cross-validation.

Carbohydrates↗

Thymus zygis subsp. Gracilis: watering level effect on phytomass production and essential oil quality.

Thymus zygis subsp. gracilis (chemotype thymol) was evaluated on the basis of its phytomass production and essential oil quality. Three different watering levels were assayed to achieve 63, 44, and 30% of the local potential evapotranspiration (Eto). According to the statistical analysis, a water supplement equivalent to 44% Eto in this cultivation area was optimal for maximum plant dry matter production and essential oil yield. Capillary GC-MS analysis of the essential oil allowed the identification of 86 volatile components. Among them, 30 are described for the first time as volatile constituents of the essential oil in this thyme subspecies and chemotype. The watering level effect on essential oil composition was noticeable, because the application of a water supplement equivalent to the 63% Eto favored the production of an essential oil richer in low molecular weight components. However, the greatest thymol concentrations were obtained under the 30 and 44% Eto watering levels.

Gas Chromatography-Mass Spectrometry↗

Enhancing the retention of phytochemicals and organoleptic attributes in muscadine grape juice through a combined approach between dense phase CO2 processing and copigmentation.

This study evaluated the phytochemical stability and organoleptic attributes of an ascorbic acid-fortified muscadine grape juice as affected by dense phase CO2 processing (DP-CO2) and addition of thyme polyphenolic cofactors (Thymus vulgaris; 1:100 anthocyanin-to-cofactor molar ratio) in efforts to prevent phytochemical losses that occur during storage of anthocyanin-containing beverages, especially in the presence of carbonyl compounds commonly produced during thermal processing and storage. DP-CO2 processing insignificantly altered initial juice phytochemical and antioxidant content, whereas thermal pasteurization reduced anthocyanins (263 mg/L), ascorbic acid (42 mg/L), soluble phenolics (266 mg/L), and antioxidant capacity (6 microM Trolox equivalents/mL). Similar trends were observed during storage, and data showed that increasing the CO2 level from 8 to 16% during DP-CO2 was instrumental in reducing juice phytochemical and antioxidant degradation. Copigmentation was instrumental in retaining higher anthocyanin, soluble phenolics, and antioxidant capacity during storage without affecting initial juice aroma and flavor characteristics. Moreover, on the basis of overall likeability scores, panelists preferred copigmented juices, which had increased juice color intensity and masked the detrimental color fading that occurred during storage, especially when compared to thermally pasteurized juices. DP-CO2 and copigmentation were effective strategies to reduce phytochemical and color deterioration that occurred in muscadine juice during storage without affecting their organoleptic attributes.

Anthocyanins↗

Anaerobic utilization of essential oils by denitrifying bacteria.

Plant volatile organic compounds are a major carbon source in nature. We studied the degradability of these substances by anaerobic microorganisms in enrichment cultures with representative essential oils as organic substrates and nitrate as electron acceptor. Lemon and pine needle oil supported microbial growth in the presence of pure oil, whereas parsley seed, camphor, sage, fennel, and mint oil supported growth only when the essential oils were dissolved in an overlying phase of 2,2,4,4,6,8,8-heptamethylnonane. Thyme oil did not support denitrification. Analyses of the microbially degraded oils revealed the disappearance of monoterpenes, of several monoterpenoids, and of methoxy-propenyl-benzenes, including apiole and myristicin. Most-probable-number determinations for denitrifying communities in sewage sludge and forest soil yielded 10(6) to 10(7) monoterpene-utilizing cells ml(-1), representing 0.7 to 100% of the total cultivable nitrate-reducing microorganisms. The utilization of essential oils together with the common occurrence of this metabolic trait are indications for an environmentally important, but currently unexplored anaerobic turnover of plant volatile organic compounds in soil.

Alcaligenes↗

Using recall to reduce false recognition: diagnostic and disqualifying monitoring.

Whether recall of studied words (e.g., parsley, rosemary, thyme) could reduce false recognition of related lures (e.g., basil) was investigated. Subjects studied words from several categories for a final recognition memory test. Half of the subjects were given standard test instructions, and half were instructed to use recall to reduce false recognition. Manipulation checks indicated that the latter instructions did elicit a recall-to-reject strategy. However, false recognition was selectively reduced only when all the words from a category could be recalled (Experiment 1). When longer categories were used, thereby minimizing exhaustive recall, a recall-to-reject strategy was ineffective at reducing false recognition (Experiment 2). It is suggested that exhaustively recalling a category allowed subjects to disqualify the lure as having occurred, analogous to recall-to-reject demonstrations in other tasks. In contrast, partially recalling a category did not help to diagnose the lure as nonstudied. These findings constrain theories of recall-based monitoring processes.

Humans↗

Evaluation of the preservative properties of Thymus vulgaris essential oil in topically applied formulations under a challenge test.

The preservative properties of thyme essential oil (3%) with a known composition were evaluated in two types of final formulations, suitable for use as pharmaceutical or cosmetic vehicles, by means of a standard challenge test proposed by the latest European Pharmacopoeia. The required preservation efficacy criteria were satisfied against the bacterial strains, against the yeast in one of the formulations, but not against the mould strain involved in this study. Interactions between the essential oil compounds and other factors present in the final formulation might have influenced the activity of this essential oil, leading to an incomplete satisfaction of the criteria.

Administration, Topical↗

Bactericidal activity of carvacrol towards the food-borne pathogen Bacillus cereus.

Carvacrol, a natural plant constituent occurring in oregano and thyme, was investigated for its bactericidal effect towards the food-borne pathogen Bacillus cereus. Carvacrol showed a dose-related growth inhibition of B. cereus. At concentration of 0.75 mmol l-1 and above, total inhibition of the growth was observed. Below this concentration, carvacrol extended the lag-phase, reduced the specific growth rate and reduced the maximum population density. Incubation for 40 min in the presence of 0.75-3 mmol l-1 carvacrol decreased the number of viable cells of B. cereus exponentially. Spores were found to be approximately 2-3 fold less sensitive to carvacrol than vegetative cells. Bacillus cereus cells showed reduced susceptibility towards carvacrol at pH 7.0 compared with different values between pH 4.5 and 8.5. The culture and exposure temperatures had a significant influence on the survival of vegetative cells. The highest death rate of cells was observed at an exposure temperature of 30 degrees C. Membrane fluidity was found to be an important factor influencing the bactericidal activity of carvacrol.

Anti-Bacterial Agents↗

Antimicrobial activity of essential oils and other plant extracts.

The antimicrobial activity of plant oils and extracts has been recognized for many years. However, few investigations have compared large numbers of oils and extracts using methods that are directly comparable. In the present study, 52 plant oils and extracts were investigated for activity against Acinetobacter baumanii, Aeromonas veronii biogroup sobria, Candida albicans, Enterococcus faecalis, Escherichia col, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serotype typhimurium, Serratia marcescens and Staphylococcus aureus, using an agar dilution method. Lemongrass, oregano and bay inhibited all organisms at concentrations of < or = 2.0% (v/v). Six oils did not inhibit any organisms at the highest concentration, which was 2.0% (v/v) oil for apricot kernel, evening primrose, macadamia, pumpkin, sage and sweet almond. Variable activity was recorded for the remaining oils. Twenty of the plant oils and extracts were investigated, using a broth microdilution method, for activity against C. albicans, Staph. aureus and E. coli. The lowest minimum inhibitory concentrations were 0.03% (v/v) thyme oil against C. albicans and E. coli and 0.008% (v/v) vetiver oil against Staph. aureus. These results support the notion that plant essential oils and extracts may have a role as pharmaceuticals and preservatives.

Anti-Bacterial Agents↗

Antimicrobial agents from plants: antibacterial activity of plant volatile oils.

The volatile oils of black pepper [Piper nigrum L. (Piperaceae)], clove [Syzygium aromaticum (L.) Merr. & Perry (Myrtaceae)], geranium [Pelargonium graveolens L'Herit (Geraniaceae)], nutmeg [Myristica fragrans Houtt. (Myristicaceae), oregano [Origanum vulgare ssp. hirtum (Link) Letsw. (Lamiaceae)] and thyme [Thymus vulgaris L. (Lamiaceae)] were assessed for antibacterial activity against 25 different genera of bacteria. These included animal and plant pathogens, food poisoning and spoilage bacteria. The volatile oils exhibited considerable inhibitory effects against all the organisms under test while their major components demonstrated various degrees of growth inhibition.

Animals↗

Inhibitory effect of essential oils on apical growth of Aspergillus fumigatus by vapour contact.

The inhibitory effect of seven essential oils on the apical growth of hyphae of Aspergillus fumigatus was studied using a bio cell tracer by vapour contact in a sealed vessel. Based on the inhibitory pattern, these essential oils were classified into three groups. The first group, composed of citron, lavender and tea tree oils, stopped the apical growth in a loading dose of 63 micrograms ml-1 air, but allowed the regrowth of the hyphae after removal of the vapour, indicating fungistatic action. The second group, consisting of perilla and lemon-grass oils, stopped the apical growth in a loading dose of 6.3 micrograms ml-1 air, and did not allow the regrowth after gaseous contact at 63 micrograms ml-1 air, indicative of fungicidal action. The third group, consisting of cinnamon bark and thyme oils, retarded the growth in a dose of 6.3 micrograms ml-1 air, stopped it in a dose of 63 micrograms ml-1 air, and incompletely suppressed regrowth of the hyphae. Gas chromatographic analysis revealed that vapours of essential oils were absorbed on fungal mycelia and agar medium most abundantly by the first group, followed by the second and third groups, reflecting the volatility of the respective groups. Suppression of the apical growth by vapour contact was ascribed to the direct deposition of essential oils on fungal mycelia, together with an indirect effect via the agar medium absorbed.

Aspergillus fumigatus↗

Antimicrobial effect of essential oils on the seafood spoilage micro-organism Photobacterium phosphoreum in liquid media and fish products.

AIMS: To evaluate the antimicrobial effect of nine essential oils (EO) on P. phosphoreum and determine the effect of oregano oil on the shelf-life of modified atmosphere-packed (MAP) cod fillets. METHODS AND RESULTS: The antimicrobial effect of EO was studied in a liquid medium and in product storage trials. Oils of oregano and cinnamon had strongest antimicrobial activity, followed by lemongrass, thyme, clove, bay, marjoram, sage and basil oils. Oregano oil (0.05%, v/w) reduced growth of P. phosphoreum in naturally contaminated MAP cod fillets and extended shelf-life from 11-12 d to 21-26 d at 2 degrees C. CONCLUSIONS: Oregano oil reduced the growth of P. phosphoreum and extended the shelf-life of MAP cod fillets. SIGNIFICANCE AND IMPACT OF THE STUDY: Mild and natural preservation using EO can extend the shelf-life of MAP seafood through inhibiting the specific spoilage organism P. phosphoreum.

Fish Products↗

In vitro effect of essential oils and isolated mono- and sesquiterpenes on Leishmania major and Trypanosoma brucei.

The effect of different essential oils as well as of isolated mono- and sesquiterpenes on the viability of bloodstream forms of Trypanosoma brucei, promastigotes of Leishmania major and human HL-60 cells was evaluated using the Almar Blue assay. Of the 12 essential oils and 8 terpenes investigated, only three essential oils, Melissa officinalis (balmmint) oil, Thymus vulgaris (thyme) oil, and Melaleuca alternifolia (tea tree) oil were about 50-fold and 80-fold more toxic to bloodstream forms of T. brucei than to HL-60 cells, respectively. Terpinen-4-ol, the main compound of the Australian tea tree oil, was even 1000-fold more toxic to trypanosomes than to the human cells. On the other hand, none of the essential oils and terpenes tested were more toxic to promastigotes of L. major than to HL-60 cells.

Animals↗

Antiviral effect of aqueous extracts from species of the Lamiaceae family against Herpes simplex virus type 1 and type 2 in vitro.

Aqueous extracts from species of the Lamiaceae family were examined for their antiviral activity against Herpes simplex virus (HSV). Extracts from lemon balm (Melissa officinalis), peppermint (Mentha x piperita), prunella (Prunella vulgaris), rosemary (Rosmarinus officinalis), sage (Salvia officinalis) and thyme (Thymus vulgaris) were screened. Their inhibitory activity against Herpes simplex virus type 1 (HSV-1), type 2 (HSV-2) and an acyclovir-resistant strain of HSV-1 (ACV (res)) was tested in vitro on RC-37 cells in a plaque reduction assay. The 50% inhibitory concentrations (IC (50)) of the extracts for HSV plaque formation were determined in dose-response studies. All test compounds showed a high antiviral activity against HSV-1, HSV-2 and ACV (res). In order to identify the mode of antiviral action, the extracts were added to the cells or viruses at different stages of infection. Both types of Herpes virus including ACV (res) were considerably neutralized after treatment with the extracts prior to infection. At maximum non-cytotoxic concentrations of the extracts, plaque formation was significantly reduced by > 90% for HSV-1 and HSV-2 and > 85% for ACV (res). In time-response studies over a period of 2 hours, a clearly time-dependent activity was demonstrated. These results indicate that the extracts affect HSV before adsorption, but have no effect on the intracellular virus replication. Therefore, the extracts exert their antiviral effect on free HSV and offer a chance to use them for topical therapeutic application against recurrent HERPES infections.

Antiviral Agents↗

In vitro inhibition of human cytochrome P450-mediated metabolism of marker substrates by natural products.

Spices, herbal and black teas, and soybean products were analyzed for their capacity to inhibit in vitro metabolism of drug marker substrates by human cytochrome P-450 (CYP) isoforms. Inhibition of drug metabolism was determined using aliquots or infusions from these products in a fluorescence-detection assay. Aliquots and infusions of all natural product categories inhibited 3A4 metabolism to some extent. Of the 26 aliquots from teas and spices further tested with 2C9, 2C19 and 2D6, many demonstrated significant inhibitory activity on the metabolism mediated by these isoforms. Black teas and herbal tea mixtures were generally more inhibitory than single-entity herbal teas. Spices and single-entity herbal teas showed species-specific isoform inhibition with sage, thyme, cloves, St John's Wort and goldenseal having the highest activity against several isoforms. Seven soybean varieties tested, as well as daidzein and genistein isolated from soybean, were found to inhibit 3A4-mediated metabolism. Genistein was found to inhibit 3A7- but not 3A5-mediated metabolism of the marker substrate. Assessment of the in vitro CYP inhibition potential for these natural products has important implications for predicting the likelihood of natural product-drug interactions if these products are taken concomitantly.

Chromatography, High Pressure Liquid↗