Aerial parts of Achillea moschata Wulfen (Asteraceae) growing wild in the Italian Rhaetian Alps were investigated to describe, for the first time, their phenolic content, as well as to characterize the essential oil. Inspection of the metabolic profile combining HPLC-DAD and ESI-MS/MS data showed that the methanol extract contained glycosylated flavonoids with luteolin and apigenin as the main aglycones. Among them, the major compound was 7-O-glucosyl apigenin. Caffeoyl derivates were other phenolics identified. The essential oil obtained by steam distillation and investigated by GC/FID and GC/MS showed camphor, 1,8-cineole, and bornylacetate as the main constituents. The antioxidant capacity of three different extracts with increasing polarity and of the essential oil was evaluated by employing ABTS¨+ and DPPH¨radical scavenging assays. The methanolic extract was the only significantly effective sample against both synthetic radicals. All samples were also tested against Gram-positive (Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa) bacterial species using the disk diffusion assay. The non-polar extracts (dichloromethane and petroleum ether) and the essential oil possessed a broad spectrum of antimicrobial activity expressed according to inhibition zone diameter (8-24 mm).
Effects of environmental growth conditions on the antioxidant capacity, total phenolic content and composition of Achillea collina Becker ex Rchb. were investigated. Methanol extracts and infusions obtained from leaves and inflorescences of plants cultivated in the Italian Alps at two different altitudes (600 and 1050 m a.s.l.) were evaluated. Infusions exhibited the highest antioxidant capacity (1/IC(50) values from 4.35 ± 0.72 to 8.90 ± 0.74), total phenolic content (from 31.39 ± 4.92 to 49.36 ± 5.70 mg gallic acid equivalents (GAE) g(-1) DW), chlorogenic acid (from 9.21 ± 1.52 to 31.27 ± 6.88 mg g(-1) DW), 3,5-di-O-caffeoylquinic acid (from 12.28 ± 3.25 to 25.13 ± 1.99 mg g(-1) DW) and 4,5-di-O-caffeoylquinic acid (from 7.38 ± 1.01 to 12.78 ± 2.61 mg g(-1) DW) content. Climate (as influenced by altitude) was shown to be the main environmental factor influencing yarrow composition and properties. Leaf extracts from the higher experimental site showed a 2-4-fold increase of chlorogenic acid level. Achillea collina can be considered as a very good source of bioactive phenolic compounds, and growing it at high altitude may constitute an effective way to significantly enhance its quality for both medicinal and nutritional uses.
Achillea moschata Wulfen, which grows in the Alps, is extensively used by local people for its medicinal properties. Two studied samples were collected, at the flowering stage, in Val Mustair (Switzerland) and Valchiavenna (Italy), respectively. The aerial parts were defatted with petroleum ether (PET) and successively extracted with dichloromethane (DCM) and methanol (MeOH). High-performance liquid chromatography and electrospray ionization-tandem mass spectrometry analyses of the methanolic extracts evidenced that flavonoids were the predominant compounds compared to phenolic acids in both samples (89.5 vs. 33.0 μg/mg DW in A. moschata Valchiavenna and 82.5 vs. 40.0 μg/mg DW in A. moschata Val Mustair). Among flavonoid derivatives, luteolin and apigenin were the predominant aglycones, free and glycosilated. The A. moschata Valchiavenna extract was characterized by apigenin as the main compound (60.4 μg/mg DW), while A. moschata Val Mustair was characterized by its derivative apigenin 7-O-glucoside (44.7 μg/mg DW). The antioxidant activity of all the obtained extracts was tested by the DPPH (2,2-diphenyl-picryl hydrazyl) and ABTS (2,21-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) methods, which showed their increasing scavenger capacity in relation to extract polarity (PET extract < DCM extract < MeOH extract). The extracts were also investigated against three Gram-positive (Bacillus cereus, Enterococcus faecalis and Staphylococcus aureus) and three Gram-negative (Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa) bacterial species using the disc diffusion assay. DMC and PET were the most active extracts (inhibition diameter: ≥12 mm).
Understanding the molecular basis of acid tolerance in the food-borne pathogen Listeria monocytogenes is important as this property contributes to survival in the food-chain and enhances survival within infected hosts. The aim of this study was to identify genes contributing to acid tolerance in L. monocytogenes using transposon mutagenesis and subsequently to elucidate the physiological role of these genes in acid tolerance. One mutant harboring a Tn917 insertion in the thiT gene (formerly lmo1429), which encodes a thiamine (vitamin B1) uptake system, was found to be highly sensitive to acid. The acid-sensitive phenotype associated with loss of this gene was confirmed with an independently isolated mutant, from which the thiT gene was deleted (∆thiT). Cells of both wild-type and ∆thiT mutant that were thiamine depleted were found to be significantly more acid sensitive than control cultures. Thiamine-depleted cultures failed to produce significant concentrations of acetoin, consistent with the known thiamine dependence of acetolactate synthase, an enzyme required for acetoin synthesis from pyruvate. As acetoin synthesis is a proton-consuming process, we suggest that the acid sensitivity observed in thiamine-depleted cultures may be owing to an inability to produce acetoin.
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