Antimicrobial and antioxidant activities of Melissa officinalis L. (Lamiaceae) essential oil

The present study describes antimicrobial and free radical scavenging capacity (RSC) together with the effects on lipid peroxidation (LP) of Melissa officinalis essential oil. The chemical profile of essential oil was evaluated by the means of gas chromatography-mass spectrometry (GC-MS) and thin-layer chromatography (TLC). RSC was assessed measuring the scavenging activity of essential oil on the 2,2-diphenyl-1-picrylhydrazyl (DPPH(*)) and OH(*) radicals. The effect on LP was evaluated following the activities on Fe(2+)/ascorbate and Fe(2+)/H(2)O(2) systems of induction. The antimicrobial activity was tested against 13 bacterial strains and six fungi. The examined essential oil exhibited very strong RSC, reducing the DPPH radical formation (IC(50) = 7.58 microg/mL) and OH radical generation (IC(50) = 1.74 microg/mL) in a dose-dependent manner. According to the GC-MS and TLC (dot-blot techniques), the most powerful scavenging compounds were monoterpene aldehydes and ketones (neral/geranial, citronellal, isomenthone, and menthone) and mono- and sesquiterpene hydrocarbons (E-caryophyllene). Very strong inhibition of LP, particularly in the Fe(2+)/H(2)O(2) system of induction (94.59% for 2.13 microg/mL), was observed in both cases, also in a dose-dependent manner. The most effective antibacterial activity was expressed on a multiresistant strain of Shigella sonei. A significant rate of antifungal activity was exhibited on Trichophyton species.     

Enhancement of polyphenol recovery from rosemary (Rosmarinus officinalis) and sage (Salvia officinalis) by enzyme-assisted ensiling (ENLAC).

The efficacy of enzyme-assisted ensiling (ENLAC) in the recovery of polyphenols from rosemary and sage was tested. Fresh rosemary and sage were chopped and ensiled in 0.5-L anaerobic jars. Treatments comprised control (no additives), 0.5% glucose and lactic acid bacteria, and 1% cellulase plus 1% hemicellulase plus pectinase. Following storage at room temperature for 45 days (experiment 1) and 26 days (experiment 2), polyphenols were extracted from the silages in ethanol either by direct blending or by cold extraction. The enzyme treatment resulted in silages with the lowest pH values, lowest fiber content, highest water-soluble sugar content, and highest polyphenol recovery; this treatment resulted in increased polyphenol recovery from rosemary and sage, by 100 and 20%, respectively. Comparison between direct blending and cold extraction revealed similar efficiency of polyphenol recovery.     

Essential oils produced by in vitro shoots of sage (Salvia officinalis L.)

In vitro shoots of sage (Salvia officinalis L.) were established under eight different hormonal supplementations and proliferated by subculture of nodal shoot segments. The respective essential oils, obtained by hydrodistillation, were composed of more than 75 compounds, 65 of which were identified. The 10 major compounds were, by order of retention time, alpha-pinene (4.1-5.4%), camphene (6-7.1%), beta-pinene (9.3-14.5%), limonene (2-2.3%), 1,8-cineole (3.6-5.6%), (-)-thujone (13.2-16.1%), (+)-isothujone (6.6-7.4%), camphor (19.8-24%), alpha-humulene (5.1-6.8%), and manool (4.2-7.7%). Notwithstanding the eight different hormonal supplementations tested, the percentage composition of the shoot essential oils were kept in a narrow range of variation. However, the type and concentration of growth regulators apparently influenced the accumulation of essential oils. The highest accumulation of essential oils and the highest shoot biomass growth were obtained with 2.0 mg/L kinetin and 0.05 mg/L 2,4-dichlorophenoxyacetic acid.     

Main agronomic-productive characteristics of two ecotypes of Rosmarinus officinalis L. and chemical composition of their essential oils

The productive potential of two different ecotypes of Rosmarinus officinalis (Cevoli and Lunigiana) cultivated in the littoral area near Pisa (northern Tuscany, Italy) and the differences in the yield and composition of the essential oils of leaves, flowers, and stems obtained from different positions of the plants were used to characterize the two ecotypes. The Cevoli ecotype plant produced the highest yield of dry matter (221 g plant-1) in comparison to the Lunigiana ecotype (72 g plant-1). There were significant differences in dry matter production of different organs of both ecotypes. The essential oil contents of Cevoli and Lunigiana ecotypes were similar. In contrast, the oil contents of the different plant parts showed marked differences. The apical part of the plant and the leaves gave the highest essential oil yields. The major difference between the oils of the two ecotypes consisted in the 1,8-cineole contents (6.6 and 37.9% in Cevoli and Lunigiana, respectively). The Cevoli ecotype was determined to be the most suitable for essential oil extraction because it was characterized by a preponderance of flowers and leaves in the apical portion. The Cevoli ecotype could be classifited as an alpha-pinene chemotype, whereas Lunigiana is a 1,8-cineole chemotype.     

Variation of the chemical profile and antioxidant behavior of Rosmarinus officinalis L. and Salvia fruticosa Miller grown in Greece

In this study, the essential oil and the phenolic composition along with the antioxidant activity of R. officinalis L. and S. fruticosa Miller, collected in Zakynthos island (Ionian Sea, Greece), were investigated. The essential oil composition of the plants was characterized by the presence of 1,8-cineole. Mean values of the antioxidant activities of rosemary and sage essential oils indicated slight differences. The antioxidant activity of sage oil was correlated with the oxygenated sesquiterpenes and diterpenes concentrations. Concerning the methanolic extracts, a close relationship between the phenolic content and the development stage during vegetative cycle of these plants was observed. The identified flavonoids, except rutin, seemed to increase with the advancement of developmental stages, while phenolic acids followed an opposite pattern. The antioxidant activity was correlated with the amount of total phenolic content.     

Influence of organic mulching and nitrogen application on essential oil yield and nitrogen use efficiency of rosemary (Rosmarinus officinalis L.)

In a field study conducted at Bangalore, India for two years (2003–2004 and 2004–2005), eight treatment combinations consisting of two variables, organic mulch (lemongrass spent material as mulch at 7.5 t ha^?1 and no mulch) and four levels of nitrogen fertilizer (0, 100, 200 and 300 kg ha^?1) were examined to observe the effect of organic mulching on nitrogen use-efficiency, and herb and essential oil yield in a multiharvested rosemary crop. The results revealed that application of lemongrass spent material as mulch increased the herb and essential oil yields in rosemary by 16.2 and 24.2%, respectively, over the non-mulched control at first harvest. Corresponding values for the regenerated crop harvest were 18.8 and 16.8%. A significant response to N was observed with 300 kg N ha^?1 in non-mulched plots compared with 200 kg N ha^?1 in mulched plots. Using lemongrass spent material as mulch, nitrogen uptake by the rosemary crop increased by 15.1% over the non-mulched control. At 200 kg N ha^?1, apparent recoveries by the crop were estimated to be 33.64% for the non-mulched control, and 37.79% with mulch. The quality of the rosemary essential oil, in terms of the concentrations of its major constituents, α-pinene, 1 : 8 cineole, camphor and verbenone, was not affected by the use of organic mulching and nitrogen fertilization, and these constituents were found to be of astandard acceptable in international trade.     

Relevance of carnosic acid concentrations to the selection of rosemary, Rosmarinus officinalis (L.), accessions for optimization of antioxidant yield

Methods were developed to identify and select accessions of rosemary, Rosmarinus officinalis (L.), producing optimum antioxidant activity. Extracts from 12 different rosemary accessions, using three solvents of varying polarity, were assayed for their antioxidant activity, and their major antioxidant compounds were identified and quantified by high-performance liquid chromatography (HPLC). Carnosic acid concentrations were correlated with (i) the free radical scavenging activity of these extracts, as measured by the 2,2-diphenyl-1-picrylhydrazyl assay (adjusted R(2) = 77.3%) and (ii) their inhibition of linoleic acid oxidation, as measured by the beta-carotene assay (adjusted R(2) = 44.1%). The correlation was broadly confirmed by the production of volatile aldehydes as measured by the hexanal assay. The variation of carnosic acid concentrations in extracts of 29 accessions, grown in field trials at three sites in England, was determined.     

Isolation and structural elucidation of two new glycosides from sage (Salvia officinalis L.)

Six compounds, 1-O-(2,3, 4-trihydroxy-3-methyl)butyl-6-O-feruloyl-beta-D-glucopyranoside, ethyl beta-D-glucopyranosyl tuberonate, p-hydroxybenzoic acid, (-)-hydroxyjasmonic acid, caffeic acid, and 4-hydroxyacetophenone 4-O-[5-O-(3, 5-dimethoxy-4-hydroxybenzoyl)-beta-D-apiofrunosyl]-(1-->2)-beta-D- glu copyranoside, were isolated from the n-butanol-soluble fraction of sage leaf extracts. Their structures were determined by spectral methods (MS, NMR, and 2D-NMR), and their antioxidant activities were measured. Among them, two new glycosides were elucidated. All of these compounds showed DPPH free radical scavenging activity at the concentration of 30 mM, and caffeic acid was the most active compound.     

Ionic interactions and salinity affect monoterpene and phenolic diterpene composition in rosemary (Rosmarinus officinalis)

In this study, we evaluated how increased cation supply can alleviate the toxic effects of NaCl on plants and how it affects essential oils (EOs) and phenolic diterpene composition in leaves of rosemary (Rosmarinus officinalis L.) plants grown in pots. Two concentrations of the chloride salts KCl, CaCl₂, MgCl₂, and FeCl₃ were used together with 100 mM NaCl to study the effects of these nutrients on plant mineral nutrition and leaf monoterpene, phenolic diterpene, and EO composition. The addition of 100 mM NaCl, which decreased K⁺, Ca²⁺, and Mg²⁺ concentrations with increasing Na⁺ in leaves, significantly altered secondary metabolite accumulation. Addition of MgCl₂ and FeCl₃ altered leaf EO composition in 100 mM NaCl–treated rosemary plants while KCl and CaCl₂ did not. Furthermore, addition of CaCl₂ promoted the accumulation of the major phenolic diterpene, carnosic acid, in the leaves. The carnosol concentration was reduced by the addition of KCl to salt‐stressed plants. It is concluded that different salt applications in combination with NaCl treatment may have a pronounced effect on phenolic diterpene and EO composition in rosemary leaves thus indicating that ionic interactions may be carefully considered in the cultivation of these species to achieve the desired concentrations of these secondary metabolites.     

Dynamics of yield components and essential oil production in a commercial hybrid sage (Salvia officinalis x Salvia fruticosa cv. Newe Ya'ar no. 4).

The fresh yields, the essential oil content, and the quality of a sage hybrid (Salvia officinalis x Salvia fruticosa, cv. Newe Ya'ar No. 4, Lamiaceae) as affected by development and harvest time were determined. Marked increases in plant height and in the number of nodes developed per plant together with a modest increase in leaf size were accompanied by dramatic increases (more than 20-fold) in the fresh yields throughout a 50-day growth period. No major changes in the essential oil content per fresh weight and its composition were detected throughout the growth period. In contrast, the compositions of the essential oils obtained from stems, as compared to leaves and leaf-primordia, had marked differences. Developmentally controlled changes in the extractives from individual leaf pairs from the same plant were also noted. In upper young leaves, the oxygenated diterpene manool and the sesquiterpene hydrocarbons alpha-humulene and beta-caryophyllene constituted up to 20%, 8%, and 4% of the total extractives, respectively. In older leaves, the abundance of these components steadily dropped to roughly half their levels in young leaves. Conversely, the proportions of the monoterpenes, particularly the ketones camphor and alpha-thujone, steadily increased with leaf position. Minor changes in the levels of other extractives were also recorded. These studies imply independent regulatory patterns for di-, sesqui-, and monoterpenes in this sage hybrid, and suggest possible agrotechnical means to obtain preferred chemical compositions of its essential oil.     

Rapid quantitative enrichment of carnosic acid from rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography

For the first time, the potent but unstable antioxidative diterpene carnosic acid could be enriched from an aqueous extract of rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography. Enrichment of carnosic acid in the foam was influenced by the pH value and the flow rate of the foam-forming gas. Efficiency was highest with diluted samples at pH 4. Under these conditions, the conversion of carnosic acid to carnosol was negligible. Transfer of carnosic acid to the foam from a standard solution in the presence of saponin as surfactive substance was similar to that from the aqueous rosemary extract.     

Azo dye-mediated regulation of total phenolics and peroxidase activity in thyme (Thymus vulgaris L.) and rosemary (Rosmarinus officinalis L.) clonal lines

Thyme (Thymus vulgaris L.) and rosemary (Rosmarinus officinalis L.) clonal lines, which were previously isolated from a heterogeneous seed population by plant tissue culture techniques, have been targeted as potential plants for phytoremediation of organic pollutants such as azo dyes and related aromatic compounds. Three thyme clonal lines and three rosemary clonal lines were tested for the ability to grow on hormone-free medium containing 0.01% of azo dye Poly S-119. The results showed that dye tolerance was associated with reduced phenolics and enhanced peroxidase activity in these clonal lines. There was a clear inverse correlation between total phenolics and peroxidase activity in these plants in response to Poly S-119. The tolerance of these clonal lines showed variations at different growing stages. These observations suggested that the peroxidase activity was inducible. Because peroxidases are involved in lignification, wound healing, aromatic compound degradation, pathogen defense, and stiffening, the results suggest that azo dye stimulated the defense response of thyme and rosemary clonal plants by increasing the peroxidase activity. Stereomicroscopic observations revealed that the azo dye was sequestered within the growing axis of the plant roots, which may also enhance the polymerization of azo dye onto the cell wall with the help of enhanced peroxidase activity.     

Essential oils from dalmatian sage (Salvia officinalis l.): variations among individuals, plant parts, seasons, and sites.

The factors affecting oil yield and quality of essential oils from Dalmatian sage (Salvia officinalis L.) are analyzed. Distillations of oils from individual plants and GC analyses revealed the presence of three chemotypes with different proportions of alpha- and beta-thujone (alpha/beta 10:1, 1.5:1, and 1:10). Different accessions could also be classified as having high (39-44%), medium (22-28%), or low (9%) total thujone contents. Flowering parts of S. officinalis had higher oil contents (1.6 versus 1.1%) and beta-pinene levels (27 versus 10%) than leaves and lower thujone levels (16 versus 31%). Major seasonal changes were found in the composition of oil distilled from a flowering type of Dalmatian sage, but oil yields from healthy, established plants did not vary greatly. Total thujone levels were lowest (25%) around flowering in spring and summer, so autumn or winter was the best harvest time to obtain oils with high thujone levels.     

Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey. Var. pectinatus (Lamiaceae)

The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.     

Genetic diversity and population structure in medicinal plant Melissa officinalis L. (Lamiaceae)

Genetic Resources and Crop Evolution - Melissa officinalis is an important medicinal plant and is extensively used by locals. This species is growing in different regions of Iran and forms several...     

Chemical composition and antibacterial activity of essential oils from Thymus spinulosus Ten. (Lamiaceae)

The chemical composition of essential oils from aerial parts of Thymus spinulosus Ten. (Lamiaceae) is reported. Four oils from plants growing in different environmental conditions were characterized by GC and GC-MS methods; the oils seem to indicate a new chemotype in the genus Thymus. Influences of soil and altitude characteristics on the essential oil composition are discussed. The oils showed antibacterial activity against Gram-positive (Staphylococcus aureus, Streptococcus faecalis, Bacillus subtilis, and Bacillus cereus) and Gram-negative (Proteus mirabilis, Escherichia coli, Salmonella typhimuium Ty2, and Pseudomonas aeruginosa) bacteria.     

Polyphenolic transmission to Segureno lamb meat from ewes' diet supplemented with the distillate from rosemary (Rosmarinus officinalis) leaves

The aim of the present work is to study whether the introduction of rosemary plant byproduct, from plant steam distillation, in daily Segurena sheep feeding allows the transfer of active antioxidant components to lamb meat, without detriment to the animal productivity. For this, 36 Segurena ewes were assigned randomly to three homogeneous groups. One group was fed a basal diet as a control and the diet of the other two groups was modified by substituting 10 or 20% of the control diet (respectively) with distilled rosemary leaves. Chromatographic analysis allowed the identification of 11 polyphenolic components previously identified in the rosemary and basal diet pellets, respectively. Among them, rosmarinic acid, carnosol, and carnosic acid were the phenolic components that had a significantly increased presence ( P < 0.05) in the lamb meat from sheep mothers fed this aromatic herb, when compared to the control group. The incorporation of this byproduct into the animal diet favored the antioxidant capacity of these lamb meat samples. Fresh meat produced on rosemary had higher total ferric reducing antioxidant power (FRAP) ( P < 0.05), greater ability to reduce ABTS*+, and lower IC50 (DPPH*) ( P < 0.05) values when compared to the control group. Because no statistically significant differences were detected among the results obtained from the lamb meat belonging to the different ewe groups fed rosemary leaf extract (10 or 20%), it can be concluded that the incorporation of distilled rosemary leaves at a rate of 10% of the ewes' diet should be enough to improve the lamb meat antioxidant status.     

Antioxidant activity of chemical components from sage (Salvia officinalis L.) and thyme (Thymus vulgaris L.) measured by the oil stability index method

A new abietane diterpenoid, 12-O-methyl carnosol (2), was isolated from the leaves of sage (Salvia officinalis L.), together with 11 abietane diterpenoids, 3 apianane terpenoids, 1 anthraquinone, and 8 flavonoids. Antioxidant activity of these compounds along with 4 flavonoids isolated from thyme (Thymus vulgaris L.) was evaluated by the oil stability index method using a model substrate oil including methyl linoleate in silicone oil at 90 degrees C. Carnosol, rosmanol, epirosmanol, isorosmanol, galdosol, and carnosic acid exhibited remarkably strong activity, which was comparable to that of alpha-tocopherol. The activity of miltirone, atuntzensin A, luteolin, 7-O-methyl luteolin, and eupafolin was comparable to that of butylated hydroxytoluene. The activity of these compounds was mainly due to the presence of ortho-dihydroxy groups. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of these compounds showed the similar result.     

Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) andOriganum vulgare L

Antilisterial activities of Thymbra capitata and Origanum vulgare essential oils were tested against 41 strains of Listeria monocytogenes. The oil of T. capitata was mainly constituted by one component, carvacrol (79%), whereas for O. vulgare three components constituted 70% of the oil, namely, thymol (33%), gamma-terpinene (26%), and p-cymene (11%). T. capitata essential oil had a significantly higher antilisterial activity in comparison to O. vulgare oil and chloramphenicol. No significant differences in L. monocytogenes susceptibilities to the essential oils tested were registered. The minimum inhibitory concentration values of T. capitata essential oil and of carvacrol were quite similar, ranging between 0.05 and 0.2 microL/mL. Antioxidant activity was also tested, the essential oil of T. capitata showing significantly higher antioxidant activity than that of O. vulgare. Use of T. capitata and O. vulgare essential oils can constitute a powerful tool in the control of L. monocytogenes in food and other industries.     

Recovery mechanism of the antioxidant activity from carnosic acid quinone,an oxidized sage and rosemary antioxidant

A solution of carnosic acid quinone, which is a radical chain-termination product having no antioxidant activity in the antioxidant reaction of carnosic acid, recovers potent antioxidant activity upon standing. The HPLC analysis of an aged solution of carnosic acid quinone revealed that several antioxidants are produced in the solution. From the time-course and quantitative analyses of the formation of the products and their structural analysis, an antioxidant mechanism from carnosic acid quinone is proposed that includes a redox reaction of carnosic acid quinone in addition to the isomerization to lactone derivatives. In the first stage of antioxidation, carnosic acid, the reduction product from carnosic acid quinone, contributes to the potent antioxidant activity of the solution. This proposed mechanism can explain one of the reasons for the strong antioxidant activity of the extract of the popular herbs sage and rosemary.