GC-MS Chemical Profiling, Biological Investigation of Three Salvia Species Growing in Uzbekistan.

Salvia is a potentially valuable aromatic herb that has been used since ancient times. The present work studied the chemical profile of three Salvia species essential oils (EO): S. officinalis, S. virgata and S. sclarea, as well as assessing their antioxidant and enzyme inhibitory activities. A total of 144 compounds were detected by GC-MS analysis, representing 91.1, 84.7 and 78.1% in S. officinalis, S. virgata and S. sclarea EOs, respectively. The major constituents were cis-thujone, 2,4-hexadienal and 9-octadecenoic acid, respectively. The principal component analysis (PCA) score plot revealed significant discrimination between the three species. The antioxidant activity of the EOs was evaluated using in vitro assays. Only S. virgata EO showed antioxidant activity in the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay (26.6 ± 1.60 mg Trolox equivalent (TE)/g oil). Moreover, this oil exhibited the highest antioxidant activity in 2,2-azino bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), cupric-reducing antioxidant capacity (CUPRAC) and ferric-reducing power (FRAP) assays in comparison with the other two EOs (190.1 ± 2.04 vs. 275.2 ± 8.50 and 155.9 ± 1.33 mg TE/g oil, respectively). However, S. virgata oil did not show any effect in the chelating ability assay, while in the PBD assay, S. officinalis had the best antioxidant activity (26.4 ± 0.16 mmol TE/g oil). Enzyme inhibitory effect of the EOs was assessed against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), tyrosinase, α-glucosidase and α-amylase. AChE enzyme was more sensitive to S. officinalis EO (4.2 ± 0.01 mg galantamine equivalent (GALAE)/g oil), rather than S. virgata EO, which was ineffective. However, S. virgata had the highest BChE effect (12.1 ± 0.16 mg GALAE/g oil). All studied oils showed good tyrosinase inhibitory activity, ranging between 66.1 ± 0.61 and 128.4 ± 4.35 mg kojic acid equivalent (KAE)/g oil). Moreover, the EOs did not exhibit any glucosidase inhibition and were weak or inefficient on amylase enzyme. Partial least squares regression (PLS-R) models showed that there is an excellent correlation between the antioxidant activity and the volatile profile when being compared to that of enzyme inhibitory activity. Thus, the studied Salvia essential oils are interesting candidates that could be used in drug discovery for the management of Alzheimer's and hyperpigmentation conditions.

Application of Five Different Chlorella sp. Microalgal Strains for the Treatment of Vegetation Waters Derived from Unconventional Oil Extractions Enriched with Citrus Byproducts.

The Mediterranean diet has, among its cornerstones, the use of olive oil for its nutraceutical and organoleptic properties. Despite the numerous merits, olive-oil mill wastewater (OMWW), which is generated by the olive-oil extraction process, is one of the most serious environmental pollutants in the Mediterranean countries. The polluting potential of OMWW is due to its high content of tannins, polyphenols, polyalcohols, pectins and lipids. In order to close the recovery cycle of a fortified citrus olive oils previously developed, we tested the ability of five microalgae of the Chlorella group (SEC_LI_ChL_1, CL_Sc, CL_Ch, FB and Idr) in lowering the percentage of total phenolic compounds in vegetation water. This was obtained with three different extraction processes (conventional, and lemon and orange peels) at three concentrations each (10%, 25% and 50%). The results showed that strains Idr, FB and CL_Sc from the Lake Massaciuccoli can tolerate vegetation water from conventional and lemon peel extractions up to 25%; these strains can also reduce the phenolic compounds within the tests. The application of microalgae for OMWW treatment represents an interesting opportunity as well as an eco-friendly low-cost solution to be developed within companies as a full-scale approach, which could be applied to obtain a fortified microalgal biomass to be employed in nutraceutical fields.

Volatilome Analyses and In Vitro Antimicrobial Activity of the Essential Oils from Five South African Helichrysum Species.

Helichrysum genus was used in folk South African medicine to treat various human disorders. As a part of our on-going research addressing the exploitation of South African plants belonging to this genus, five species were investigated for their volatile and antimicrobial activities. The volatile organic compounds (VOCs) and the essential oils (EOs) were analysed by gas chromatography mass spectrometry (GC-MS). Microdilution was the method used for assessing both antimycotic and antibacterial activities, which was also tested by Kirby-Bauer agar disc diffusion. Total monoterpenes (TMs) dominated the VOCs of four species (H. trilineatum (70.6%), H. edwardsii (79.3%), H. cooperi (84.5%), and H. pandurifolium (57.0%)). H. cooperi and H. edwardsii EOs showed the predominance of TMs (68.2% and 84.5%, respectively), while H. pandurifolium and H. trilineatum EOs were characterized by the prevalence of TSs (86.5% and 43.6%, respectively). H. odoratissimum EO evidenced a similar amount of both TMs (49.5%) and TSs (46.4%). Microsporum canis was more sensitive to these EOs. The lowest minimum inhibitory concentration (MIC) was observed with H. pandurifolium and H. edwardsii EOs (0.25%). H. pandurifolium and H. trilineatum had a good effect on Staphylococcus aureus (MIC 5%). These findings open new perspectives for the exploitation of these natural compounds for application in cosmetics and pharmaceutics.

Helichrysum araxinum Takht. ex Kirp. grown in Italy: volatiloma composition and in vitro antimicrobial activity.

In the present work the composition of biogenic volatile organic compounds (BVOCs) and the essential oil (EO) of Helichrysum araxinum Takht. ex Kirp. aerial parts, together with the antimicrobial activity, were investigated. The results showed the prevalence of sesquiterpene hydrocarbons in both spontaneous emissions as well as in the EO. The main compounds of BVOCs were γ-curcumene (10.7%), γ-muurolene (9.2%), and ß-selinene (8.5%). This latter constituent also showed a similar amount in the EO and represented the most abundant compounds together with α-selinene (8.0%). It is Interesting to note the same percentage of monoterpene hydrocarbons (MHs) in both the aroma profile and the EO (18.0%) with the same most abundant compounds: ß-pinene (6.3% in BVOCs vs. 5.1% in EO, respectively) and limonene (4.5% in VOCs vs. 4.9% in EO, respectively). With regard to the antimycotic activity, the EO showed to be inactive against the tested strains, while a moderate antibacterial activity was shown against Staphylococcus isolates.

Chemical Composition and in Vitro Cytotoxic Screening of Sixteen Commercial Essential Oils on Five Cancer Cell Lines.

The in vitro cytotoxic activity on human cancer cell lines of sixteen commercial EOs such as Aloysia citriodora, Boswellia sacra, Boswellia serrata, Cinnamomum zeylanicum, Cistus ladanifer, Citrus × aurantium, Citrus limon, Citrus sinensis, Cymbopogon citratus, Foeniculum vulgare, Illicium verum, Litsea cubeba, Satureja montana, Syzygium aromaticum, Thymus capitatus and Thymus vulgaris was performed using the MTT reduction assay. The screening was carried out on human cancer cells of breast adenocarcinoma (MCF7, T47D and MDA-MB-231), chronic myelogenous erythroleukemia (K562) and neuroblastoma cell lines (SH-SY5Y). C. zeylanicum and L. cubeba EOs were the most active on almost all the cell lines studied and thus could be promising as an anticancer agent. These two species showed a difference in their composition even though they belong to the Lauraceae family. Almost 57 % of the true cinnamon composition was made of (E)-cinnamaldehyde, while L. cubeba showed citral as the major compound (68.9 %). The K562 cells were the most sensitive to these oils with an IC50 ranging from 5.2 parts-per million (ppm) (C. zeylanicum) to 11.1 ppm (L. cubeba). The latter oil also showed an important cytotoxicity on MDA-MB-231 (13.4 ppm).

Volatilomic Analysis of Four Edible Flowers from Agastache Genus.

Volatilomes emitted from edible flowers of two species of Agastache (A. aurantiaca (A.Gray) Lint & Epling, and A. mexicana (Kunth) Lint & Epling) and from two hybrids (Agastache 'Arcado Pink' and Agastache 'Blue Boa') were investigated using a solid-phase microextraction technique as well as the extraction of its essential oils. Oxygenated monoterpenes were almost always the predominant class (>85%) of volatile organic compounds (VOCs) in each sample of A. aurantiaca, A. 'Blue Boa' and A. mexicana, with the exception of A. 'Arcado Pink' (38.6%). Pulegone was the main compound in A. aurantiaca (76.7%) and A. 'Blue Boa' (82.4%), while geranyl acetate (37.5%) followed by geraniol (16%) and geranial (17%) were the principal ones in A. mexicana. The essential oil composition showed the same behavior as the VOCs both for the main class as well as the major constituent (pulegone) with the same exception for A. mexicana. Total soluble sugars, secondary metabolites (polyphenols, flavonoids and anthocyanins) and antioxidant activity were also investigated to emphasize the nutraceutical properties of these edible flowers.

In Vitro Activity of Essential Oils against Saprolegnia parasitica.

Saprolegnia spp. water molds severely impact fish health in aquaculture, fish farms and hobby fish tanks colonizing mature and immature stages of fishes, as well as eggs. Considering that there are no drugs licensed for treating and/or control the organism, efficient and environmental low-impact methods to control these oomycetes in aquaculture are needed. The aim of the present report was to evaluate the in vitro sensitivity of Saprolegnia parasitica to essential oils (EOs) from Citrus aurantium L., Citrus bergamia Risso et Poiteau, Citrus limon Burm. f., Citrus paradisi Macfad, Citrussinensis Osbeck, Cinnamomum zeylanicum Blume, Cymbopogon flexuosum (Nees ex Steud.) Watson, Foeniculum vulgare Mill., Illicium verum Hook.f., Litsea cubeba (Lour.) Pers., Origanum majorana L., Origanum vulgare L., Pelargonium graveolens L'Hér., Syzygium aromaticum Merr. & L.M.Perry, and Thymus vulgaris L., by microdilution test. The most effective EOs assayed were T. vulgaris and O. vulgare, followed by C. flexuosum, L. cubeba and C. bergamia. These EOs could be of interest for controlling Saprolegnia infections. Nevertheless, further safety studies are necessary to evaluate if these products could be dispersed in tank waters, or if their use should be limited to aquaculture supplies.

Activity of selected essential oils on spoiling fungi cultured from Marzolino cheese.

Microscopic fungi can be present on a variety of foodstuff, including cheese. They can be responsible for fungal spoilage, causing sensory changes making food unacceptable for human consumption, and posing severe health concerns. Furthermore, some of these organisms are able to resist antimicrobial preservatives provided for by law. Antifungal activity of 15 chemically defined EOs, alone and in mixture, were checked by a microdilution test against isolates of Penicillium funiculosum and Mucor racemosus cultured from rinds of Marzolino, a typical Italian fresh pecorino cheese. Origanum vulgare yielded the lowest MIC values, followed by Salvia sclarea, Ocimum basilicum and Cymbopogon citratus, while Citrus paradisi and Citrus limon were not active. All mixtures showed antifungal activity at lower concentration with respect to MIC values of each EO component, when not in combination. This study is the first to describe the setting up of EOs mixtures to limit spoiling moulds.

Stonebrood and chalkbrood in Apis mellifera causing fungi: in vitro sensitivity to some essential oils.

Aim of the present study was to evaluate the in vitro antimycotic activity of 17 chemically defined essential oils (EOs) both alone and as a mixture, against agents responsible for stonebrood caused by Aspergillus flavus, and chalkbrood caused by Ascosphaera apis in European honeybees. Cinnamomum zeylanicum yielded the lowest MIC value against A. flavus, but was not effective against A. apis, while Litsea cubeba and Pelargonium graveolens appeared to be effective against all checked fungi. Aspergillus niger showed the lower sensitivity. Two mixtures composed by L. cubeba, C. zeylanicum and Cymbopogon flexuosus (M1) and by L. cubeba, C. zeylanicum, P. graveolens and C. flexuosus (M2), respectively, were tested, both resulting effective. The components of M1 showed a synergistic effect. The use of mixtures allowed to decrease the total amount of EOs. The use of these products could be of interest for an alternative natural approach in honeybee disease management.

Sub-inhibitory stress with essential oil affects enterotoxins production and essential oil susceptibility in Staphylococcus aureus.

Fourteen wild strains of Staphylococcus aureus positive for gene sea were tested for enterotoxins production and the minimum inhibitory concentration of Leptospermum scoparium, Origanum majorana, Origanum vulgare, Satureja montana and Thymus vulgaris essential oils (EOs) were determined. After this trial, bacteria stressed with sub-inhibitory concentration of each EO were tested for enterotoxins production by an immunoenzymatic assay and resistance to the same EO. Oregano oil exhibited the highest antibacterial activity followed by manuka and thyme oils. After the exposure to a sub-inhibitory concentration of EOs, strains displayed an increased sensitivity in more than 95% of the cases. After treatment with oregano and marjoram EOs, few strains showed a modified enterotoxins production, while 43% of the strains were no longer able to produce enterotoxins after treatment with manuka EO. The results obtained in this study highlight that exposure to sub-inhibitory concentration of EO modifies strains enterotoxins production and EOs susceptibility profile.

Taxonomy of prickly juniper (Juniperus oxycedrus group): A phytochemical-morphometric combined approach at the contact zone of two cryptospecies.

Based on different essential oil composition paralleling different genotypes, Juniperus deltoides was recently segregated from Juniperus oxycedrus. Despite a clear phytochemical and molecular differentiation, J. deltoides resulted not clearly morphologically discernible from J. oxycedrus, so that it was defined as a cryptospecies. Italy represents the contact zone of their distribution, but the ranges of the two species are not sufficiently known, due to unsatisfactory morphological characterisation. To further complicate the picture, a third closely related species (ecotype), J. macrocarpa, occurs all across the Mediterranean coasts. After a preliminary phytochemical analysis to ascertain the (chemo-)identities of the studied populations, we performed a morphometric investigation to test the degree of morphological distinctiveness among the taxa. According to our analysis, some character (e.g. leaf mucro length, leaf width, seed-cone size and seed size) resulted useful to discriminate these cryptic taxa. Finally, based on these characters, an extensive revision of herbarium specimens allowed us to redefine the distribution pattern of the investigated species in the Central Mediterranean area.

Traditional Mediterranean plants: characterization and use of an essential oils mixture to treat Malassezia otitis externa in atopic dogs.

Several plants extracts from Mediterranean countries are traditionally employed in skin troubles both in humans and in animals. Malassezia pachydermatis is a lipophylic yeast responsible for otitis externa and dermatitis in dogs and for cutaneous and systemic disease in humans. Five mixtures of essential oils obtained from Mediterranean plants (Citrus paradisi, Salvia sclarea, Ocimum basilicum, Rosmarinus officinalis, Citrus limon, Anthemis nobilis, Lavandula hybrida and Thymus vulgaris) provided with antifungal and/or anti-inflammatory action assayed in vitro, were tested in vivo versus M. pachydermatis to treat once daily for 2 weeks 25 atopic dogs with Malassezia otitis externa. Mixture composed by C. limon 1%, S. sclarea 0,5%, R. officinalis 1%, A. nobilis 0,5% yielded excellent results in all treated dogs. Despite of clinical resolution after all treatments the number of blastospores did not decrease. This study confirms recent findings suggesting a multifactorial alternative approach for the management of canine Malassezia otitis.