Flowering phenophases influence the antibacterial and anti-biofilm effects of Thymus vulgaris L. essential oil.

BACKGROUND: Essential oils are becoming increasingly popular in medicinal applications because of their antimicrobial effect. Thymus vulgaris L. (Lamiaceae) is a well-known and widely cultivated medicinal plant, which is used as a remedy for cold, cough and gastrointestinal symptoms. Essential oil content of thyme is responsible for its antimicrobial activity, however, it has been reported that the chemical composition of essential oils influences its biological activity. In order to explore flowering phenophases influence on the chemical composition of thyme essential oil and its antibacterial and anti-biofilm activity, plant materials were collected at the beginning of flowering, in full bloom and at the end of flowering periods in 2019. METHODS: Essential oils from fresh and dried plant materials were distilled and analyzed with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The antibacterial activity was performed by broth microdilution and thin layer chromatography-direct bioautography (TLC-DB) assays and the anti-biofilm effect by crystal violet assay, respectively. Scanning electron microscopy was applied to illustrate the cellular changes of bacterial cells after essential oil treatment. RESULTS: Thymol (52.33-62.46%) was the main component in the thyme essential oils. Thyme oil distilled from fresh plant material and collected at the beginning of flowering period exerted the highest antibacterial and anti-biofilm activity against Haemophilus influenzae, H. parainfluenzae and Pseudomonas aeruginosa. CONCLUSION: The different flowering periods of Thymus vulgaris influence the antibacterial and anti-biofilm activity of its essential oils, therefore, the collection time has to be taken into consideration and not only the full bloom, but the beginning of flowering period may provide biological active thyme essential oil.

Quality Evaluation of Light- and Dark-Colored Hungarian Honeys, Focusing on Botanical Origin, Antioxidant Capacity and Mineral Content.

Melissopalynology, antioxidant capacity and mineral and toxic element contents were analyzed in eight types of Hungarian honeys. Based on color, two groups were distinguished: light honeys comprised acacia, amorpha, phacelia and linden honeys; while dark honeys included sunflower, chestnut, fennel and sage honeys, with 100 to 300 and 700 to 1500 mAU, respectively. The unifloral origin of each sample was supported using pollen analysis. The absorbance of honey correlated positively with antioxidant capacity determined by three different methods (TRC, DPPH, ORAC), and also with mineral content. The exception was the light amber linden honey with significantly higher K content and antiradical activity than other light honeys. The Mn, Zn and Fe contents were the highest in chestnut, sunflower and fennel honeys, respectively. The black meadow sage honey performed best regarding the content of other elements and antioxidant activity. The concentrations of several toxic elements were below the detection limit in the samples, indicating their good quality. The principal component analysis (PCA) revealed correlations between different antioxidant assays and minerals, and furthermore, confirmed the botanical authentication of the honeys based on the studied parameters. To our best knowledge, the present study is the first to provide a complex analysis of quality parameters of eight unifloral Hungarian honeys.

Soil Humus, Iron, Sulphate and Magnesium Content Affect Nectar Traits of Wild Garlic (Allium ursinum L.).

Recent studies revealed that from various ecological factors influencing nectar yield and quality of a plant, soil properties can be as important as microclimatic features. To date, few studies have investigated the relationship of soil characters to nectar traits of bee pollinated plants growing in natural associations. Our study intended to reveal which soil properties had the most powerful impact on nectar variables of wild garlic (Allium ursinum L.). Specimens were collected from fourteen habitats in two different years, and were potted in their original soil under the same climatic conditions. Nectar volumes and sugar concentrations were measured and soil samples were analysed for fourteen parameters. Statistical analyses revealed that the number of nectar producing Allium flowers, as well as the nectar volume and sugar content of nectar in individual flowers were influenced by both year and habitat. The humus, iron and sulphate content of soil showed negative correlation with the number of flowers producing nectar; total nectar volumes were negatively correlated with humus and iron content, but positively affected by magnesium content of the soil. Our results suggest that in addition to the effect of microclimatic factors, certain soil properties can have significant impact on nectar traits.

Chemical Composition and Antiproliferative Effect of Essential Oils of Four Solidago Species (S. canadensis, S. gigantea, S. virgaurea and S.×niederederi).

There is a growing interest in the research of natural products with potent activity in the prevention and treatment of diseases. Thereby, essential oil phytochemicals have demonstrated broad bioactivities. This study highlights the chemical composition of essential oils obtained from the leaves and inflorescences of diverse species of the genus Solidago (Solidago canadensis, Solidago gigantea, Solidago virgaurea and Solidago×niederederi), also known as Goldenrod, through GC/MS analysis, and their antiproliferative activity on three human tumor cell lines (MDA-MB 231, A375 and HCT116) by the MTT assay. The most represented chemical classes in the essential oils were oxygenated sesquiterpenes, sesquiterpene hydrocarbons, oxygenated monoterpenes and monoterpene hydrocarbons, with a remarkable contribution of compounds such as germacrene D (3.89-19.51 %), α-pinene (3.59-18.82 %), bornyl acetate (3.30-12.73 %) and caryophyllene oxide (3.07-9.18 %). Chemical quali- and quantitative differences were noticed between the leaves and flowers of the same species, as well as between the studied species. The MTT assay showed that the essential oils from the leaves were more active than those from the flower ones. The essential oil having the highest impact on cell viability was that from S.×niederederi (IC50 =12.93, 6.72, and 6.82 µg/ml), followed by S. virgaurea (IC50 =13.39, 7.96, and 8.36 µg/ml) and finally S. gigantea (IC50 =18.04, 5.94, and 8.10 µg/ml) on MDA-MB 231, A375 and HCT116 cell lines, respectively. The essential oil from S. canadensis showed the lowest activity (IC50 =29.33, 12.63, and 18.03 µg/ml, respectively). Taken together, these findings highlight the Solidago species as good sources of cytotoxic compounds. Further studies are encouraged on the in vivo effects and safety of these products.

What happens in Europe stays in Europe: apparent evolution by an invader does not help at home.

Some invasive plant species rapidly evolve greater size and/or competitive ability in their nonnative ranges. However, it is not well known whether these traits transfer back to the native range, or instead represent genotype-by-environment interactions where traits are context specific to communities in the new range where the evolution occurred. Insight into transferability vs. context specificity can be tested using experiments performed with individuals from populations from the native and nonnative ranges of exotic invasive species. Using a widespread invasive plant species in Europe, Solidago gigantea, we established reciprocal common garden experiments in the native range (Montana, North America; n = 4) and the nonnative range (Hungary, Europe; n = 4) to assess differences in size, vegetative shoot number, and herbivory between populations from the native and nonnative ranges. In a greenhouse experiment, we also tested whether the inherent competitive ability of genotypes from 15 native and 15 invasive populations differed when pitted against 11 common native North American competitors. In common gardens, plants from both ranges considered together produced five times more biomass, grew four times taller, and developed five times more rhizomes in the nonnative range garden compared to the native range garden. The interaction between plant origin and the common garden location was highly significant, with plants from Hungary performing better than plants from Montana when grown in Hungary, and plants from Montana performing better than plants from Hungary when grown in Montana. In the greenhouse, there were no differences in the competitive effects and responses of S. gigantea plants from the two ranges when grown with North American natives. Our results suggest that S. gigantea might have undergone rapid evolution for greater performance abroad, but if so, this response does not translate to greater performance at home.

Anti-Haemophilus Activity of Selected Essential Oils Detected by TLC-Direct Bioautography and Biofilm Inhibition.

Essential oils (EOs) are becoming increasingly popular in medical applications because of their antimicrobial effect. Direct bioautography (DB) combined with thin layer chromatography (TLC) is a screening method for the detection of antimicrobial compounds in plant extracts, for example, in EOs. Due to their lipophilic character, the common microbiological assays (etc. disk diffusion) could not provide reliable results. The aim of this study was the evaluation of antibacterial and anti-biofilm properties of the EO of cinnamon bark, clove, peppermint, thyme, and their main components against Haemophilus influenzae and H. parainfluenzae. Oil in water (O/W) type Pickering nano-emulsions stabilized with silica nanoparticles from each oil were prepared to increase their water-solubility. Samples with Tween80 surfactant and absolute ethanol were also used. Results showed that H. influenzae was more sensitive to the EOs than H. parainfluenzae (except for cinnamon bark oil). In thin layer chromatography-direct bioautography (TLC-DB) the ethanolic solutions of thyme oil presented the best activity against H. influenzae, while cinnamon oil was the most active against H. parainfluenzae. Pickering nano-emulsion of cinnamon oil inhibited the biofilm formation of H. parainfluenzae (76.35%) more efficiently than samples with Tween80 surfactant or absolute ethanol. In conclusion, Pickering nano-emulsion of EOs could inhibit the biofilm production effectively.

Polar Constituents, Essential Oil and Antioxidant Activity of Marsh Woundwort (Stachys palustris L.).

Stachys palustris, well-known as marsh woundwort, is a perennial herb growing in wet environments of Europe. Its tubers, leaves and seeds are eaten raw or cooked. Alike other Stachys species, the plant is also used as a traditional remedy. Despite S. palustris has been consumed for centuries, little is known about its chemical constituents. In this work the main secondary metabolites of S. palustris from Hungary and France have been analysed. From the plant ethanolic extracts, ethanoid glucosides, isoscutellarein derivatives, caffeoyl-quinic acids and iridoids have been isolated and structurally characterized by NMR. The essential oils were analysed by GC/MS and showed (E)-phytol, fatty acids and carbonylic compounds as the most abundant compounds. The radical scavenging capacity of plant ethanolic extracts, as evaluated by the DPPH assay, was noteworthy, with IC50 values of 92.08 - 105.42 µg/ml.

Kainate postconditioning restores LTP in ischemic hippocampal CA1: onset-dependent second pathophysiological stress.

Postconditioning can be induced by a broad range of stimuli within minutes to days after an ischemic cerebral insult. A special form is elicited by pharmacological intervention called second pathophysiological stress. The present study aimed to evaluate the effects of low-dose (5 mg/kg) kainate postconditioning with onsets 0, 24 and 48 h after the ischemic insult on the hippocampal synaptic plasticity in a 2-vessel occlusion model in rat. The hippocampal function was tested by LTP measurements of Schaffer collateral-CA1 pyramidal cell synapses in acute slices and the changes in density of Golgi-Cox-stained apical dendritic spines. Postconditioning 0 and 24 h after ischemia was not protective, whereas 48-h-onset postconditioning resulted in the reappearance of a normal spine density (>100,000 spines) 3 days after ischemia, in parallel with the long-term restoration of the damaged LTP function. Similar, but somewhat less effects were observed after 10 days. Our data clearly demonstrate the onset dependence of postconditioning elicited by a subconvulsant dose of kainate treatment in global ischemia, with restoration of the structural plasticity and hippocampal function.

Oxaloacetate decreases the infarct size and attenuates the reduction in evoked responses after photothrombotic focal ischemia in the rat cortex.

A traumatic brain injury or a focal brain lesion is followed by acute excitotoxicity caused by the presence of abnormally high glutamate (Glu) levels in the cerebrospinal and interstitial fluids. It has recently been demonstrated that this excess Glu in the brain can be eliminated into the blood following the intravenous administration of oxaloacetate (OxAc), which, by scavenging the blood Glu, induces an enhanced and neuroprotective brain-to-blood Glu efflux. In this study, we subjected rats to a photothrombotic lesion and treated them after the illumination with a single 30-min-long administration of OxAc (1.2 mg/100 g, i.v.). Following induction of the lesion, we measured the infarct size and the amplitudes of the somatosensory evoked potentials (SEPs) as recorded from the skull surface. The photothrombotic lesion resulted in appreciably decreased amplitudes of the evoked potentials, but OxAc administration significantly attenuated this reduction, and also the infarct size assessed histologically. We suggest that the neuroprotective effects of OxAc are due to its blood Glu-scavenging activity, which, by increasing the brain-to-blood Glu efflux, reduces the excess Glu responsible for the anatomical and functional correlates of the ischemia, as evaluated by electrophysiological evoked potential (EP) measurements.