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Human populations in various regions across the world exploit the medicinal properties of plants to treat a wide variety of diseases. Areas with both high rates of endemic taxa and persisting traditional uses of the local botanical resources are key sites for the investigation of Traditional Botanical Knowledge (TBK). Commonly, in these areas, information regarding the medicinal properties of native plants has been transmitted orally from generation to generation, however, a rapid decline in this knowledge has been observed, which can be attributed to socio-economic changes in recent years. The Mediterranean basin is one such site, where human history is intimately entwined with nature. The unique geographical situation and unrivaled environmental heterogeneity of the area, have allowed both the development of diverse civilizations as well as providing the basis for the evolution of extraordinary biodiversity. The Mediterranean basin can therefore be considered a global hotspot of endemic vascular plants, and of traditional knowledge of medicinal and aromatic species. This study researches the historical subregion of Marmilla (central-southern Sardinia, Italy), which was chosen because of its specific cultural and demographic characteristics: i.e., prolonged isolation and extreme longevity of the inhabitants of the area. Semi-structured interviews were conducted with 145 people from the region, and 137 medicinal plants belonging to 62 families were identified, of which around 57,3% were taxa exclusive to the Mediterranean Basin. Findings showed that the most used parts of the plant were the leaves (49%), while as far as preparations are concerned, decoction (50%) was the most used to prepare medicinal formulations, making this the highest number of medico-botanical taxa reported in a study carried out in Sardinia using a similar methodology. In addition, this study contributes towards preventing the loss of TBK by documenting the medicinal traditions, passed down orally for centuries, in the words of the participants, shedding new light on the traditional knowledge of the inhabitants of the island. The findings lay the foundations for future applied studies in the fields of phytotherapy and phytochemical investigation., Competing Interests: The authors declare no conflict of interest.

The aim of this study is to demonstrate the antifungal, anti-inflammatory and anti-migratory potential of the essential oil of Teucrium scordium subsp. scordioides (Schreb.) Arcang, a plant widely used in traditional medicine in Sardinia. The oil was rich in germacrene D (25.1%), δ-cadinene (12.9%) and alloaromadendrene (11.3%). The yeast Cryptococcus neoformans and the dermatophytes Trichophyton rubrum , T. mentagrophytes var. interdigitale and Epidermophyton floccosum were the most susceptible fungi to the action of the oil. In lipopolysaccharide (LPS)-stimulated macrophages, the oil was able to decrease nitric oxide production by ca. 30% at 1.25 μL/mL, without affecting cell viability. In the scratch wound assay, it allowed for ca. 36% of wound closure after 18 h, thus showing anti-migratory properties. Overall, this study highlights the potential of this species to mitigate fungal infections associated with an inflammatory response. Furthermore, we also reported for the first time its anti-migratory capacity, thus suggesting anticancer properties.

Chemical composition determination and in vitro antibacterial effect of Essential Oil (EO) of Laurus nobilis leaves grown North West Algerian conditions were investigated. Antibacterial activity was evaluated by the technique of the paper disc diffusion. Extractions were carried out with Clevenger apparatus and EO compositions were investigated by gas chromatography coupled to mass spectrometry (GC-MS) analysis. The oxygenated monoterpenes (MO) were the class with the highest contents, represent 59% of the EO. A total of twenty-seven of these compounds were identified, the major components were 1,8-cineole (30.1%), α-terpynil acetate (21.6%), methyl eugenol (16.9%) of the total oil. The highest antibacterial power of the EO tested was recorded in Streptococcus faecalis and Staphylococcus aureus with the zone diameter of inhibition 13.6 and 11.2 mm respectively and a minimal inhibitory concentration (MIC) of 0.25 mg/mL for both these two strains.

Santolina insularis is a Sardinian endemism that is widely used in traditional medicine. The essential oil was obtained with a yield of 2.7% and is mainly characterized by β-phellandrene (22.6%), myrcene (11.4%) and artemisia ketone (7.6%). The oil significantly reduced NO production without affecting macrophages viability. In addition, it substantially inhibited the expression of two key pro-inflammatory enzymes, iNOS and COX-2 (71% and 25% at 0.54 mg/mL). Furthermore, the oil had a promising antifungal activity being Cryptococcus neoformans (MIC = 0.13 mg/mL) and the majority of dermatophytes (MIC = 0.13 mg/mL) the most susceptible fungi. Moreover, it significantly decreased the yeast-to-hypha transition (80% inhibition at 0.13 mg/mL) on C. albicans long before showing inhibitory effects. Overall, these results show that S. insularis could be applied in the treatment of fungal infections and associated inflammatory response.

The chemical composition of two populations of Teucrium capitatum L. one from the coastline and the other one from the mountainous area of Sardinia (Italy) was assessed. Two chemotypes were identified: limonene/α-pinene/(E)-nerolidol chemotype predominant in the coastline, and limonene/α-pinene/α-trans-bergamotene/humulene epoxide II chemotype common in plants growing in the mountainous area. In addition, our results showed that the sample growing in the coastline had a more promising antifungal activity. Furthermore, this sample was highly effective in inhibiting C. albicans germ tube formation, at doses well below its MIC. Overall, this study shows that the edaphoclimatic characteristics play an important role on the essential oil composition and biological activity of Teucrium capitatum L.

The genus Teucrium L. (Lamiaceae) is a genus growing in mild climate zones, particularly in the Mediterranean Basin and Central Asia. It is represented by 11 taxa in Sardinia (Italy), living commonly in sunny habitats. In this study, the following eight Sardinian Teucrium taxa were selected, and the essential oils (EOs), obtained by stem distillation, were analyzed by GC-FID and GC-MS: T. capitatum subsp. capitatum , T. chamaedrys subsp. chamaedrys , T. flavum subsp. glaucum , T . marum , T. massiliense , T. scordium subsp. scordioides , T. scorodonia , and T. subspinosum . The comprehensive analyses led to the identification of 87 constituents representing the majority of the volatile compounds. Significant differences, both qualitative and quantitative, were observed between the taxa. Overall, monoterpenes and sesquiterpenes characterized all Teucrium EOs: T. capitatum subsp. capitatum and T. flavum subsp. glaucum revealed the highest content of monoterpene hydrocarbons, while in the other Teucrium taxa sesquiterpene hydrocarbons prevailed. Worthy of note, diterpenes were found only in T . marum and T. subspinosum , whereas T. massiliense was rich in non-terpenic oxygenated compounds. To the best of our knowledge, this is the first comprehensive report on the chemical composition of EOs obtained from Sardinian Teucrium species.

Plant products provide an alternative and successful source of lead compounds for the pharmaceutical industry. The present study was aimed to evaluate, in cell-based assays, the antiviral properties of essential oils obtained from plants that commonly grow in Sardinia, Italy, against a broad spectrum of RNA/DNA viruses. The essential oils of Helichrisumitalicum (Roth) G. Don ssp. microphyllum (Willd.) Nyman, Laurus nobilis L., Mirtuscommunis L., Pistacia lentiscus L., Salvia officinalis L., Saturejathymbra L., Lavandula angustifolia Mill., Foeniculum vulgare Mill., and Eucalyptus globulus Labill. were extracted by hydrodistillation and analyzed by gas chromatography mass spectrometry (GC-MS). Interestingly, the essential oil of Salvia officinalis showed moderate activity against bovine viral diarrhea virus (BVDV), an enveloped RNA virus belonging to the Flaviviridae family. BVDV is responsible for several clinical manifestations in bovines, including respiratory, gastroenteric, and reproductive diseases, with a significant economic impact. With the aim to individuate the constituent of the Salvia officinalis responsible for the biological activity, we tested the major components of the oil: camphene, β-pinene, limonene, 1,8-cineole, cis-thujone, camphor, (E)-caryophyllene, and α-humulene. Here, we describe α-humulene as an active component that is non-cytotoxic and active against BVDV (EC 50 = 36 µM). Its antiviral effects were evaluated using virucidal cytopathic effect inhibition and viral yield reduction assays. This is the first scientific report showing the anti BVDV effects of Salvia officinalis essential oil and α-humulene as the main active component.

This work reports the results and the comparison concerning the chemical and biomolecular analyses and the antifungal activity of three wild Pistacia species (Anacardiaceae) from Sardinia. Volatile oils from leaves and twigs of Pistacia x saportae, Pistacia lentiscus and Pistacia terebinthus were characterised using GC-FID and GC-MS techniques and tested against some fungal strains. Two DNA nuclear regions (ITS and 5S-rRNA-NTS) were amplified through PCR technique and sequenced. The three **Pistacia have similar chemical profile, although there are some important quantitative differences. The analysis of ITS and 5S-rRNA-NTS regions, reveals a species-specific nucleotide variation among the three **taxa. This method could emerge as a powerful tool for the species identification, especially because the discrimination of these three **taxa appears difficult for non-expert botanists. Concerning the antifungal activity, P. lentiscus and P. x saportae show the highest activity against Cryptococcus neoformans, with a MIC value of 0.32 μL/mL.

Introduction: Among athletes, an increasing use of nicotine via smokeless tobacco has been reported. However, there are currently unanswered questions about whether the use by athletes is due to nicotine's addictive properties and/or to benefits in physical and cognitive performance (e.g., decision-making). In this original article we reported about, (i) snus-induced reinforcing effects among snus-user athletes (Survey) and (ii) the effects of snus on the Iowa Gambling Task (IGT) in snus-user skiers (Experimental study). IGT is an experimental neuropsychological task that has been previously used on athletes and addicts to test decision-making. Methods: Survey: data were collected with the modified Cigarette Evaluation Questionnaire (mCEQ) that was administered to 61 winter sport athlete snus-users in Northern Italy. Experimental study: IGT data included: amount of money earned, number of choices from advantageous and disadvantageous decks and overall net score. Eighteen male snus-users were tested under satiety or after 12-h abstinence conditions according to a crossover design. Results: Survey: the comparison between occasional vs. regular snus-users showed a statistically significant difference in satisfaction ( P = 0.0088), calm ( P = 0.0252), and enjoyment ( P = 0.0001) mCEQ items suggesting a snus intake/effect relationship. Experimental study: significantly higher IGT net scores were found during the first 20 choice cards after abstinence vs. satiety conditions ( P = 0.0024). Conclusion: In the Survey, regular snus use induces greater satisfaction and psychological reward than occasional use. In the Experimental study, snus intake might produce an early and transient cognitive improvement on IGT in abstinent snus-users, presumably acting as a withdrawal relief.

Sarcopoterium spinosum fruits have been used to get extracts of different nature; two fixed oils were obtained by means of supercritical fluid extraction (SFE) with CO 2 at 250 bar and 40°C and using n-hexane in a Soxhlet extraction (SE) apparatus. Aqueous solutions: an aromatic water (AW) and a residual water (RW) were obtained by hydrodistillation (HD). In the RW, following have been identified: quercetin glucuronide, luteolin 7-O-glucuronide, isorhamnetin 3-O-glucuronide, quercetin sulfate and quercetin. Among all tested plant extracts, the RW had the highest content of polyphenol (378 mg GAE/g of weight) and of flavonoids (26 mg QE/g of weight), and the highest antioxidant activity, comparable to that of Trolox. It was also the most active extract of this series (IC 50  = 0.292 mg/mL) in the tyrosinase activity assays performed with L-3,4-dihydroxyphenylalanine (L-DOPA) as substrate.

In the present work the chemical compositions, measured by GC and GC-MS, of the essential oils obtained by hydrodistillation from leaves of Pistacia terebinthus collected in Bizerte (Tunisia) and Baunei (Italy) are reported. Both essential oils possessed high content of monoterpene hydrocarbons (86.3% and 90.9%, respectively), being α-pinene (62.4 vs. 35.0)%, camphene (3.0 vs. 2.4)%, β-pinene (12.1 vs. 4.5)%, terpinolene (1.7 vs. 35.2)% and β-phellandrene (3.8 vs. 4.5)% the main components. The Tunisian essential oil exhibited higher antifungal activity than the Italian one. Cryptococcus neoformans and the majority of dermatophyte strains showed more sensitivity to the Tunisian oil, when compared to Candida strains, in particular Trichophyton rubrum, Microsporum canis and Epidermophyton floccosum, with MIC and MLC values in the range (0.16-0.32) μL/mL. The results obtained support the use of the oil from Tunisia for the treatment of dermatophytosis.

Leaf essential oils of 11 samples from Ruta montana, Ruta chalepensis subsp. latifolia and Ruta chalepensis subsp. angustifolia collected from two sites have been studied by GC/MS that allowed to identify 22, 25 and 12 components, respectively. The variability in the composition of essential oils among species and seasons was very high, and hence the existence of different chemotypes has been inferred. The major components were 2-undecanone, (E) caryophyllene, 2-nonanone and 2-methyloctyl acetate. Multivariate analysis of the 11 samples suggest that the main factors capable to influence the chemical composition of leaf essential oils was both the collection period and the geographical origin. 2-Undecanone seems to be the chemotaxonomic marker of Ruta genus. Furthermore, the formation of distinct clusters and sub-clusters at high distance also supports that the patterns of distribution of chemical constituents among seasons can be used in delimiting species and infraspecific taxa within the species complexes.

Two fern species Asplenium adiantum-nigrum L. and Asplenium trichomanes L. collected from the Kroumiria region (Northwest of Tunisia) were individually submitted to hydrodistillation in a Clevenger type apparatus. Volatile organic compounds were identified by GC-MS and GC-FID. Thus, 35 compounds were identified in A. adiantum-nigrum essential oil accounting for 77.5% of the whole constituents dominated by palmitic acid (34.5%); however, only 29 volatiles were identified in A. trichomanes showing a high amount of phytol, an odorous diterpene alcohol, representing 14.4% of the total oil contents. The total phenolic content and the antioxidant effects of crude extracts from both pteridophytes were determined using Folin-Ciocalteu and 2,2'-diphenyl-1-picrylhydrazyl free radical-scavenging assays, respectively. A. adiantum-nigrum ethyl acetate extract is shown to be lower in total phenolic contents (49.3 mg gallic acid equivalent/g) than similar extract from A. trichomanes (55.4 mg GAE/g)., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The aim of this work was to formulate Santolina insularis essential oil-loaded nanocarriers, namely Penetration Enhancer containing Vesicles (PEVs), evaluate the physico-chemical features and stability, and gain insights into their ability to deliver the oil to the skin. S. insularis essential oil was obtained by steam distillation, and was predominantly composed of terpenes, the most abundant being β-phellandrene (22.6%), myrcene (11.4%) and curcumenes (12.1%). Vesicles were prepared using phosphatidylcholine, and ethylene or propylene glycol were added to the water phase (10% (v/v)) to improve vesicle performances as delivery systems. Vesicles were deeply characterized by light scattering, cryogenic transmission electron microscopy and small/wide-angle X-ray scattering, the results showing polyhedral, faceted, unilamellar vesicles of ∼115 nm in size. The presence of the glycols improved vesicle stability under accelerated ageing conditions, without changes in size or migration phenomena (e.g. sedimentation and creaming). Confocal laser scanning microscopy images of pig skin treated with S. insularis formulations displayed a penetration ability of PEVs greater than that of control liposomes. Moreover, all formulations showed a marked in vitro biocompatibility in human keratinocytes. These findings suggest that the nanoformulation may be of value in enhancing the delivery of S. insularis essential oil to the skin, where it can exert its biological activities., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Cynomorium coccineum L. is a non-photosynthetic plant, spread over Mediterranean countries, amply used in traditional medicine. The aim of this study was to evaluate for the first time the antifungal activity of its extracts. The antifungal activity was evaluated using the macrodilution method against Candida spp., Cryptococcus neoformans and dermatophyte strains. The methanolic extract was very active against C. neoformans, Candida guilliermondii and Candida krusei, with minimal inhibitory concentrations (MIC) values of 0.025 mg/mL. This extract is more active than fluconazole against C. krusei H9. The influence of methanolic extract on the dimorphic transition in Candida albicans was also studied through the germ tube inhibition assay. More than 60% of filamentation was inhibited at a concentration of 1/4 MIC. These results are preliminary and further studies are needed to an eventual use of C. coccineum methanolic extract in the treatments of candidiasis and cryptococcosis.

The objectives of this study were to chemically characterise and evaluate the antioxidant potential of the essential oil from Teucrium flavum L. subsp. flavum growing spontaneously in Tunisia. The volatile oil was extracted by hydrodistillation of the aerial parts in a Clevenger type apparatus. Forty constituents were identified via GC and GC-MS analysis. β-caryophyllene (32.5%) and α-humulene (17.8%) were the most abundant components. The evaluation of free radical scavenging activity using stable DPPH free radical showed that the volatile oil exhibits a moderate antioxidant activity and reduces DPPH to 50% at EC50 value of 1230 μg mL(-1).

The anti-inflammatory activity of two extracts from the aerial parts of Ledum palustre has been reported. The volatile oil was obtained by supercritical fluid extraction (SFE) and the essential oil by hydrodistillation (HD). The oils were analysed by gas chromatography-mass spectrometry to monitor their composition. Both extracts shared as main compound (41.0-43.4%) ledol (23.3-26.7%) and ascaridole (15.1-4.5%). The anti-inflammatory activity was evaluated by the subcutaneous carrageenan injection-induced hind paw oedema. The treated animals received essential oil (SFE and HD), the reference group received ketoprofen or piroxicam and the control group received NaCl 0.9%. A statistical analysis was performed by the Student t-test. The results show that L. palustre essential oil enhanced a significant inhibition of oedema (50-73%) for HD oil and (52-80%) for SFE oil. These results were similar to those obtained with piroxicam (70%) and ketoprofen (55%).

This article reports the results on the composition and antifungal effect of volatile extracts obtained from the aerial parts of Sardinian wild fennel (Foeniculum vulgare Mill.), by supercritical fluid extraction (SFE) and by hydrodistillation (HD). The extracts were analysed by gas chromatography-mass spectrometry for qualitative composition and gas chromatography-flame ionisation detector to establish the percentage of constituents. The main components were fenchone (7.1% vs. 8.8%), estragole (34.9% vs. 42.6%) and (E)-anethole (24.6% vs. 43.4%) in the SFE and HD extract, respectively. Minimum inhibitory concentrations (MICs) were measured according to the reference Clinical and Laboratory Standards Institute (CLSI) broth macrodilution protocols. Minimum lethal concentrations were determined by subsequent subculturing of the same cell suspensions in solid medium. The essential oil was more active against Candida albicans, whereas the supercritical fluid extract possesses higher activity against Candida guillermondii and Cryptococcus neoformans, with MIC values of 0.32 μL/mL.

This article reports the results concerning the qualitative and quantitative composition of the volatile oil obtained from the aerial parts of Tanacetum vulgare, collected in Lithuania, by supercritical fluid extraction (SFE) with CO2 at 90 bar and 40 °C and by hydrodistillation (HD). GC-MS and GC-FID analysis revealed that the oils mainly consisted of 1,8-cineole, trans-thujone, cis-chrysanthenol, borneol and myrtenol. Oils obtained by HD displayed, with respect to the SFE volatile oil, a higher amount of hydrocarbon monoterpenes. The hydrodistilled oil was particularly active against dermatophyte strains and Cryptococcus neoformans, with MIC values in the range of 0.16-0.64 μL/mL. The oil revealed a significant inhibitory effect on germ tube formation in Candida albicans at sub-inhibitory concentrations. At the concentration of 1/16 MIC, the inhibition of filamentation was more than 80% in comparison to untreated control cells.

This study investigates whether the essential oil prepared from Carum carvi seeds exhibits antiulcerogenic activity. Its volatile oil was obtained by supercritical fluid extraction (SFE) and by hydrodistillation. The essential oils were analysed by GC-MS to monitor their composition. The chemical analysis revealed that the essential oils extracted under SFE conditions had high carvone and limonene contents. The antiulcerogenic activity was evaluated by the HCl/ethanol method, which causes injury to the gastric mucosa. Three treated groups received the essential oil (100-300 mg/kg). The reference group received omeprazole (30 mg/kg) and the control group received NaCl. After 30 min, all groups were treated with HCl/EtOH for gastric ulcer induction. The results show C. carvi essential oil enhanced a significant inhibition of 47%, 81% and 88%, respectively, for three doses of essential oil used, which was similar to that induced by omeprazole (95%) (p < 0.005).

Aerial parts of Apium nodiflorum collected in Portugal and Italy were submitted to hydrodistillation; also a supercritical fluid extract was obtained from Italian plants. The extracts were analyzed by GC and GC/MS. Both essential oils, obtained from Portuguese and Italian plants, posses high content of phenylpropanoids (51.6 vs. 70.8%); in the former, the percentage split in myristicin (29.1%) and dillapiol (22.5%), whereas in the latter, the total percentage is only of dillapiol (70.8%). The co-occurrence of myristicin and dillapiol is frequent because dillapiol results from enzymatic methoxylation of myristicin. Antimicrobial activity of phenylpropanoids has been patented, what suggest the potential of plants with high amounts of these compounds. Minimal inhibitory concentration (MIC) and minimal lethal concentration, determined according to NCCLS, were used to evaluate the antifungal activity of the essential oils against yeasts, Aspergillus species and dermatophytes. Essential oils exhibited higher antifungal activity than other Apiaceae against dermatophytes, with MIC ranging from 0.04 to 0.32 μl/ml. These results support the potential of A. nodiflorum oil in the treatment of dermatophytosis and candidosis.

Unlabelled: Isolation of volatile and fixed oils from nutmeg have been obtained by supercritical fractioned extraction with carbon dioxide. Extraction experiments were carried out at pressures of 90 and 250 bar and temperature of 40 °C. The extraction step performed at 90 bar produced a volatile fraction mainly formed by myristicin (32.8%), sabinene (16.1%), α-pinene (9.8%), β-pinene (9.4%), β-phellandrene (4.9%), safrole (4.1%) and terpinen-4-ol (3.6%). The oil yield relative to this step of the process was 1.4% by weight of the charge. The last extraction step at 250 bar produced a butter-like material (nutmeg butter). The yield of this step was 14.4% by weight. The most represented fatty acids of fixed oil from nutmeg were 14:0 (79.2%), 18:1 n-9 (7.4%) and 16:0 (6.1%), and in particular the unsaturated fatty acids 18:1 n-9 averaged 32.96 μg/mg of oil. The level of myristicin in the nutmeg essential and fixed oils was also directly quantified by reversed HPLC-DAD. Moreover, the essential oil obtained from nutmeg, as well as myristicin, showed a significant in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells)., Practical Application: In this study, the chemical characterization and the anticancer activity of nutmeg oils obtained by supercritical extraction with carbon dioxide were investigated. This is important for their potential application in food and pharmaceutical industries., (© 2012 Institute of Food Technologists®)

This study is outlined to probe the chemical composition of essential oil and in vitro antioxidant activity of Juniperus oxycedrus ssp. macrocarpa (S. & m.) Ball. and Juniperus oxycedrus L. ssp. rufescens (L. K.) berries, collected from four sites, according to their maturity phase. The chemical composition of the hydrodistilled essential oil was analysed by GC-MS. Forty-eight compounds were identified, accounting for approximately 79.8-98.9% of the oil. The main constituents were α-pinene, germacrene D, myrcene, abietadiene and cis-calamenene, their mean percentage vary according to their phenological stage. The antioxidant activity of the samples was determined by the ABTS and DPPH radical scavenging activities. Hawaria essential oil extracted from mature berries showed the highest antioxidant capacity.

The topical antiinflammatory activity of the essential oil of Myrtus communis L. was studied using croton oil induced ear edema and myeloperoxidase (MPO) activity in mice, and cotton pellet induced granuloma, and serum tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in rats. On topical application, the oil exhibited a significant decrease in the ear edema as well as MPO activity. The oil also inhibited cotton pellet-induced granuloma and serum TNF-alpha and IL-6. It can be concluded that the essential oil of Myrtus communis reduces leukocyte migration to the damaged tissue and exhibits antiinflammatory activity.

This study is designed to examine the chemical composition and antioxidant activity of the essential oil of Juniperus phoenicea L. ripe and unripe berries. GC and GC/MS analyses resulted in the detection of 42 components representing approximately 96.50-99.57% of the oils. Major components of the oils were α-pinene (58.61-77.39%), camphene (0.67-9.31%), δ-3-carene (0-10.01%) and trans-verbenol (0-5.24%). Antioxidant activities were determined by two different test systems, DPPH and ABTS radical scavenging activities. In both systems ripe berries exhibited better activity potential than the unripe ones.

Satureja thymbra L. is well known in Italy by the popular name of "Santoreggia sarda". It grows only in Sardinia and nowadays it is restricted to the slope of the Colle San Michele in Cagliari. The composition of the aromatic extracts obtained by supercritical CO2 and by hydrodistillation and their antifungal activity is reported. The collected extracts were analyzed by GC-FID and GC-MS methods. No significant differences were observed in the composition of the volatile extracts depending on the extraction method. The results showed the presence of thymol, gamma-terpinene, beta-caryophyllene, p-cymene, carvacrol and borneol as main components. The minimal inhibitory concentration (MIC) and the minimal lethal concentration (MLC) were used to evaluate the antifungal activity of the oils against Candida albicans, C. tropicalis, C. krusei, C. guillermondii, C. parapsilosis, Cryptococcus neoformans, Trichophyton rubrum, T. mentagrophytes, T. mentagrophytes var. interdigitale, Trichophyton rubrum, T. verrucosum, Microsporum canis, M. gypseum, Epidermophyton floccosum, Aspergillus niger, A. fumigatus and A. flavus. The volatile extracts revealed a wide-spectrum antifungal activity. They were fungicidal and similarly potent against yeasts, dermatophyte and Aspergillus stains, with MICs ranging from 0.16 to 0.32 pL x mL(-1).

In this study, flowering aerial parts of wild Achillea millefolium growing on the Mediterranean coast (Sardinia Island, Italy) and on the Atlantic coast (Portugal- Serra de Montemuro) were used as a matrix for supercritical extraction of volatile oil with CO2 (SFE). The collected extracts were analyzed by GC-FID and GC-MS methods and their composition were compared with that of the essential oil isolated by hydrodistillation. A strong chemical variability in essential oils depending on the origin of the samples was observed. The results showed the presence of two type oils. The Italian volatile extracts (SFE and essential oil) are predominantly composed by alpha-asarone (25.6-33.3%, in the SFE extract and in the HD oil, respectively), beta-bisabolene (27.3-16.6%) and alpha-pinene (10.0-17.0%); whereas the main components of the Portuguese extracts are trans-thujone (31.4-29.0%), trans-crhysanthenyl acetate (19.8-15.8%) and beta-pinene (1.2-11.1%). The minimal inhibitory concentration (MIC) and the minimal lethal concentration (MLC) were used to evaluate the antifungal activity of the oils against Candida albicans, C. tropicalis, C. krusei, C. guillermondii, C. parapsilosis, Cryptococcus neoformans, Trichophyton rubrum, T. mentagrophytes, T. mentagrophytes var. interdigitale, T. verrucosum, Microsporum canis, M. gypseum, Epidermophyton floccosum, Aspergillus niger, A. fumigatus and A. flavus. The oils showed the highest activity against dermatophyte strains, with MIC values ranging from 0.32-1.25 microL mL(-1).

In this study, we report the preliminary data on the chemical composition of Yemeni Schinus molle L. volatile oil obtained by supercritical extraction with carbon dioxide (40°C and 90 bar), SFE, and by hydrodistillation (HD). The composition of the volatile oil has been analysed by GC and GC-MS. The content of the major constituents in the oils from leaves varied in the following ranges: germacrene D 3.7% in SFE and 16.7% in HD; β-caryophyllene 19.1% in SFE and 13.5% in HD. The amount of monoterpenes constituted 4%, in all the analysed samples, while the number of sesquiterpenes was 44% in supercritical and 67% in HD oil. Some compounds were not identified by GC-MS and it will require further analysis using other analytical techniques.

The essential oils and supercritical CO2 extracts of wild Daucus carota L. subsp. carota from two different sites in Tunisia were investigated. The main components of the essential oil of the flowering and mature umbels with seeds from Sejnane were eudesm-7(11)-en-4-ol (8.2 - 8.5%), carotol (3.5 - 5.2%), sabinene (12.0 -14.5%), a-selinene (7.4 - 8.6) and 11-alpha-(H)-himachal-4-en-1-beta-ol (12.7 - 17.4%), whereas the oils from Tunis were predominantly composed of elemicin (31.5 - 35.3%) and carotol (48.0 - 55.7%). The antimicrobial activity of the essential oils were assayed by using the broth dilution method on Escherichia coli ATCC 35218 and Staphylococcus aureus ATCC 43300, and clinical strains of Candida albicans and C. tropicalis 1011 RM. The MIC values obtained were all > 2.5% (v/v).

This article reports the composition of the essential oil from the leaves of Stachys yemenensis. The essential oil was extracted by supercritical CO₂ (90 bar; 40 °C) and its chemical composition was determined by gas chromatography and gas chromatography-mass spectrometry. The major components of the sample were α-phellandrene (13.9%), β-phellandrene (11.7%), elemol (12.0%), spathulenol (6.7%), β-eudesmol (5.0%), α-eudesmol (4.75%) and squalene (4.8%). On the exhausted matrix, deprived of the volatiles, we carried out a high-pressure (250 bar) treatment for the extraction of squalene (49.7%). The antimicrobial activity of the essential oils has been assayed by using the broth dilution method on two American Type Culture Collection (ATCC) strains, Escherichia coli ATCC 35218 and Staphylococcus aureus ATCC 43300, and two clinical strains, Candida albicans and Candida glabrata.

Isolation of volatile concentrates from leaves, flowers and fruits of Vitex agnus-castus L. have been obtained by supercritical extraction with carbon dioxide. The composition of the volatile concentrates has been analysed by GC/MS. In all plant organs, the extracts are composed chiefly of alpha-pinene, sabinene, 1,8-cineole, alpha-terpinyl acetate, (E)-caryophyllene, (E)-beta-farnesene, bicyclogermacrene, spathulenol and manool. The main difference observed was in the content of sclarene, which was not present in the samples from flowers or fruits. To complete the investigation, a comparison with the hydrodistilled oil has been carried out. The minimal inhibitory concentration (MIC) and the minimal lethal concentration were used to evaluate the antifungal activity of the oils against dermatophyte strains (Trichophyton mentagrophytes, Microsporum canis, T. rubrum, M. gypseum and Epidermophyton floccosum). Antifungal activity of the leaf essential oil was the highest, with MIC values of 0.64 microL mL(-1) for most of the strains.

This study examined the supercritical fluid extraction of wheat germ oil. The effects of pressure (200-300 bar at 40 degrees C) and extraction time on the oil quality/quantity were studied. A comparison was also made between the relative qualities of material obtained by SFE and by organic solvent extraction. The extracts were analyzed for alpha-tocopherol and polyunsaturated fatty acid content. The maximum wheat germ oil yield at about 9% was obtained with supercritical carbon dioxide extraction at 300 bar, while fatty acid and alpha-tocopherol composition of the extracts was not remarkable affected by either pressure or the extraction method.

Isolation of essential oil from dried and ground leaves of Laurus nobilis L. from Algeria and Tunisia were obtained with two different methods: by means of classical hydrodistillation (HD) and by using carbon dioxide in the supercritical state (SFE). The effect of extraction pressure on the yield and composition of the L. nobilis volatile oil is examined. The best extraction conditions were as follows: pressure, 9.0 MPa; temperature, 323.15 K; 4 h of extraction and carbon dioxide flow, Phi = 1.2 kg h(-1). Waxes were entrapped in the first separator set at 9.0 MPa and 263.15 K. The oil was recovered in the second separator working at 2.0 bar and 288.15 K. GC/MS analysis of the leaves' volatile oil revealed that it mainly consisted of: 1,8-cineole, linalool, alpha-terpinyl acetate, methyl eugenol and sabinene. The comparison with the hydrodistilled oil did not reveal any big difference. The collection of samples at different extraction times during supercritical extraction allowed us to monitor the change of the oil composition. On the matrice exhaust owing to a SFE at 9.0 MPa, one further extraction at 323.15 K and 24.0 MPa was performed to obtain a mixture of compounds with higher molecular weight. Finally, the antibacterial and antifungal activity of the various extracts has been assayed.