José G. Barroso

Antimicrobial and antioxidant properties of some commercial essential oils

The essential oil composition of Cananga odorata, Boswellia thurifera, Cymbopogon citratus, Marjorana hortensis, Ocimum basilicum, Rosmarinus officinalis, Cinnamomum zeylanicum and Citrus limon was analysed by GC and GC–MS, and their antimicrobial and antioxidant activity tested. Twenty-five different genera of bacteria and one fungal species were used in this study as test organisms. These included animal and plant pathogens, food poisoning and spoilage bacteria and the spoilage fungus Aspergillus niger. The volatile oils exhibited considerable inhibitory effect against all the tested organisms. The oils also demonstrated antioxidant capacities, comparable with α-tocopherol and butylated hydroxytoluene (BHT). The method adopted in this study was the modified thiobarbituric acid reactive species (TBARS) assay. The antioxidant activity was carried out under different conditions by using egg yolk and rat liver in the absence and presence of the radical inducer 2,2′-azobis (2-amidinopropane) dihydrochloride (ABAP).

Antimicrobial activity of essential oils isolated from Portuguese endemic species of Thymus

Aims: Thymus species are wild species mostly found in the arid lands of Portugal. Possible antimicrobial properties of Thymus essential oils have been investigated. The chemical composition of the essential oils and the antimicrobial activity of Thymus mastichina (L) L. subsp. mastichina, T. camphoratus and T. lotocephalus from different regions of Portugal were analysed.

In vitro evaluation of antioxidant activity of essential oils and their components

Clove and nutmeg essential oils were analysed by GC and GC–MS. These oils, together with 16–18 components found to be present, were tested for antioxidant properties at final concentrations of 0.05–2.5×104 ppm in an egg yolk-based thiobarbituric acid reactive substances (TBARS) assay and also undiluted in a β-carotene agar diffusion assay. Both the essential oils and the components tested in the TBARS assay demonstrated some degree of antioxidant activity. Only the clove oil, the nutmeg oil, eugenol and terpinolene demonstrated any ability to inhibit the oxidative bleaching of the β-carotene agar. The ability of the oil components to inhibit malondialdehyde formation, and therefore lipid peroxidation, in the TBARS assay, yet apparently to possess no activity in the β-carotene agar diffusion assay, demonstrates the importance in the screening of plant material for bioactivity of using a bank of assays in vitro before assigning bioactivities. By using a number of assays, not only should the number of false positives and negatives be greatly reduced, but evidence pertaining to the mechanism of action may be obtained. Copyright

Portuguese Thymbra and Thymus Species Volatiles: Chemical Composition and Biological Activities

Thymbra capitata and Thymus species are commonly known in Portugal as thyme and they are currently used as culinary herbs, as well as for ornamental, aromatizing and traditional medicinal purposes. The present work reports on the state of the art on the information available on the taxonomy, ethnobotany, cell and molecular biology of the Portuguese representatives of these genera and on the chemotaxonomy and antibacterial, antifungal and antioxidant activities of their essential oils and other volatile-containing extracts.

Plectranthus madagascariensis: Morphology of the Glandular Trichomes, Essential Oil Composition, and Its Biological Activity

Glandular and nonglandular trichomes are spread over the vegetative and reproductive organs of Plectranthus madagascariensis. Two morphologically distinct types of glandular trichomes (capitate and peltate) are described. Capitate trichomes have an ovoid unicellular head and a short stalk cell or an elongated two- to three-celled stalk slightly enlarged at the base. Peltate trichomes, which show in vivo a characteristic orange-to-reddish color, are large, flattened-to-depressed in the center, and often appear as a ring of minute gems. They are composed of a large head with eight glandular cells arranged in a single layer. An unusual kind of capitate trichomes, reported for the first time in Lamiaceae, occurs typically restricted to the calyx. These trichomes possess a two- or three-celled stalk and a long, unicellular conical, glandular head. The several types of trichomes differ in the secretion process The essential oils of P. madagascariensis, isolated by hydrodistillation and by distillation-extraction separately from flowers and from leaves collected during the flowering and the vegetative phases of the plant were analyzed by gas chromatography and gas chromatography-mass spectrometry (GC and GC-MS). The main component of the oils was a diterpene, 6,7-dehydroroyleanone, isolated as orange-to-reddish crystals, which represented 28%, 87%, and 41% of the oils from the flowers and from the leaves collected during the flowering and vegetative phases, respectively. The essential oils of P. madagascariensis showed bactericidal activity against Bacillus subtilis, Micrococcus sp., Staphylococcus aureus, and Yersinia enterocolitica, and revealed a modest antioxidant activity.

Composition and antioxidant activity of Thymus vulgaris volatiles: Comparison between supercritical fluid extraction and hydrodistillation

Supercritical fluid extraction (SFE) of the volatile oil from Thymus vulgaris L. aerial flowering parts was performed under different conditions of pressure, temperature, mean particle size and CO(2) flow rate and the correspondent yield and composition were compared with those of the essential oil isolated by hydrodistillation (HD). Both the oils were analyzed by GC and GC-MS and 52 components were identified. The main volatile components obtained were p-cymene (10.0-42.6% for SFE and 28.9-34.8% for HD), gamma-terpinene (0.8-6.9% for SFE and 5.1-7.0% for HD), linalool (2.3-5.3% for SFE and 2.8-3.1% for HD), thymol (19.5-40.8% for SFE and 35.4-41.6% for HD), and carvacrol (1.4-3.1% for SFE and 2.6-3.1% for HD). The main difference was found to be the relative percentage of thymoquinone (not found in the essential oil) and carvacryl methyl ether (1.0-1.2% for HD versus t-0.4 for SFE) which can explain the higher antioxidant activity, assessed by Rancimat test, of the SFE volatiles when compared with HD. Thymoquinone is considered a strong antioxidant compound.

Effect of ectomycorrhizal fungi on survival and growth of micropropagated plants and seedlings of Castanea sativa Mill.

Abstract Four ectomycorrhizal fungi (Amanita muscaria,Laccaria laccata,Piloderma croceum and Pisolithus tinctorius) were used to produce mycorrhiza on seedlings and micropropagated plants of Castanea sativa in vitro. Pisolithus tinctorius was most effective in colonizing roots of both micropropagated plants and seedlings. A. muscaria and L. laccata only colonized a few feeder roots of some plants and Piloderma croceum did not form mycorrhizas. Mycorrhization of micropropagated plants increased survival and growth during weaning.

Essential oils from Azorean Laurus azorica.

The essential oils isolated from leaves of ten and from unripe berries of eight populations of Laurus azorica (Seub.) Franco, collected on five islands of the Azorean archipelago, were analysed by GC and GC-MS. All oil samples were dominated by their monoterpene fraction (60-94%), alpha-pinene (15-37%) and 1,8-cineole (12-31%) being the main components of the leaf oils, while trans-beta-ocimene (27-45%) and alpha-pinene (12-22%) were the main components of the oils from the berries. The sesquiterpene fractions of the oils ranged from 3 to 17% and the main components were beta-caryophyllene (traces-8%) and beta-elemene (traces-3%) both in the leaf and berry oils. Some phenylpropanoid components were also present, in total amounting to 17%, trans-cinnamyl acetate (215% of the leaf oils) being the main component of this fraction. Cluster analysis of the enantiomeric composition of alpha- and beta-pinene in the oils from the leaves clearly showed two groups, one constituted by the two populations growing on the island S. Jorge, and the other constituted by the remaining populations.

Composition of the essential oil of Hypericum foliosum Aiton from five Azorean islands

Eight collective samples of terminal cymose inflorescences of Hypericum foliosum Aiton were collected on five islands of the Azores (S. Jorge. S. Miguel, Pico, Terceira and Faial). The essential oils isolated by hydrodistillation to estimate the oil yields, and by distillation-extraction to determine the percentage composition of the oils, were analysed by GC and GC-MS. The oil yield varied between 0.10% and 0.25% and the main components (> 5%) were n-nonane (29-73%), limonene (7-46%), terpinolene (1-19%), β-caryophyllene (1-7%) and β-pinene (0.3-6%). With the exception of one oil sample from S. Jorge, cluster analysis showed a high degree of similarity between the populations studied.

Chemical polymorphism of the essential oils from populations of Thymus caespititius grown on the island S. Jorge (Azores)

The compositions of the essential oils obtained from ten populations of Thymus caespititius grown on S. Jorge (Azores) were studied by GC and GC-MS. All the oils analysed were dominated by their monoterpene fraction (69-9%). The sesquiterpene fraction was rather small (4-17%) and consisted mainly of oxygen-containing compounds (3-13%). Enantiomeric ratio of sabinene and alpha-terpineol was investigated in the oils from some populations. (+)-Sabinene was detected with high enatiomeric purity (96 99%). Although ( +)-alpha terpineol was the predominant enantiomer, its purity varied considerably (66-97%). Cluster analysis of all identified components grouped the oils into four main clusters that corresponded with their major components: alpha-terpineol (43-68%), carvacrol (32-52%), thymol (44-58%) and sabinene (41%). The essential oil from Th. caespititius showed a clear chemical polymorphism, that was particularly evident among four populations growing along a 200 m distance at Pico Verde.