Impact of the Extraction Method on the Chemical Composition and Antioxidant Potency of Rosmarinus officinalis L. Extracts.

Rosmarinus officinalis L. is a dietary source that produces polyphenols as secondary metabolites. These natural compounds with potent antioxidant abilities are increasingly recommended as a supplement to inhibit oxidative stress. In the current work, we evaluated the impact of the extraction method on the chemical composition of R. officinalis extract, especially on the content of carnosic (CA) and rosmarinic (RA) acids using UPLC-MS-DAD as well as on their antioxidant potency. Four extracts of Tunisian rosemary were obtained from non-conventional extraction techniques:ultrasound-assisted extraction (UAE),supercritical extraction (SFE) and UAE and SFE combined ((UAE-SFE(I), UAE-SFE(II)). The UAE exhibited the best total phenolic compounds (i.e., 85.27 mg GAEg-1), the highest content of CAand RA and the strongest antioxidant abilities (i.e., IC50 = 0.13 mg/mL and EC50 = 0.93 mg/mL for DPPH scavenging test and iron reducing power ability assay). The evaluation of antioxidant activity of UAE inhuman skin fibroblast (HS-68) cell line was carried out after the induction of oxidative stress. The results determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed a strong protective effect against H2O2oxidative stress induction in cells pretreated with UAE. The obtained results allow us to give new insight about the effect of the extraction method on the chemical composition and biological activities of the extract and the importance of the choice of the most appropriate processing technique to prepare rosemary extract with a high antioxidant potency and protective effect against oxidative stress.

Antioxidant Bioactivity of Extracts from Beach Cast Leaves of Posidonia oceanica (L.) Delile.

The marine environment is a generous source of biologically active compounds useful for human health. In 50 years, about 25,000 bioactive marine compounds have been identified, with an increase of 5% per year. Peculiar feature of algae and plants is the production of secondary metabolites, such as polyphenols, synthesized as a form of adaptation to environmental stress. Posidonia oceanica is a Mediterranean endemic and dominant seagrass and represents a biologically, ecologically and geologically important marine ecosystem. Within this study, methanolic and ethanolic extracts were generated from fresh and dried Posidonia oceanica leaves, with the aim to employ and valorize the beach cast leaves. The best yield and antioxidant activity (polyphenols content equal to 19.712 ± 0.496 mg GAE/g and DPPH IC50 of 0.090 µg/µL.) were recorded in 70% ethanol extracts (Gd-E4) obtained from leaves dried for two days at 60 °C and ground four times. HPLC analyses revealed the presence of polyphenols compounds (the most abundant of which was chicoric acid) with antioxidant and beneficial properties. Bioactive properties of the Gd-E4 extracts were evaluated in vitro using fibroblast cells line (HS-68), subjected to UV induced oxidative stress. Pre-treatment of cells with Gd-E4 extracts led to significant protection against oxidative stress and mortality associated with UV exposure, thus highlighting the beneficial properties of antioxidants compounds produced by these marine plants against photo damage, free radicals and associated negative cellular effects. Beach cast leaves selection, processing and extraction procedures, and the in vitro assay results suggested the potentiality of a sustainable approach for the biotechnological exploitation of this resource and could serve a model for other marine resources.

Green biotechnology for valorisation of residual biomasses in nutraceutic sector: Characterization and extraction of bioactive compounds from grape pomace and evaluation of the protective effects in vitro.

The grape pomace (GP) is an important by-product of winemaking, accounting for about 13-25% of the grapes processed. The aim of this work was to investigate the nutritional and antioxidant composition of GP obtained from Nero d'Avola (NA) grape, one of the most important indigenous varieties in Italy, in order to verify application in nutraceutics. Seeds and skin of the GP were studied for their nutritional and antioxidants composition, fatty acid and polyphenols profile, bioactives properties in vitro, by gravimetric, spectrophotometric and chromatographic techniques. The results showed that NAGP seeds are rich in the beneficial polyunsaturated fatty acids and that the polyphenols extracted from the GP skin present a strong antiradical and antiproliferative activity, attested also in vitro, in human skin fibroblast (HS-68) and in hepatoma cell line (Hep-G2). Obtained results underline the possibility to employ this residual biomass for nutraceuticals application, contributing also to increase the sustainability.

Dietary administration effects of exopolysaccharide from potential probiotic strains on immune and antioxidant status and nutritional value of European sea bass (Dicentrarchus labrax L.).

The use of biological immunostimulants is considered a valuable practice to improve culture conditions in aquaculture sector that may help to increase production and maintain healthy environment. We undertook this study in order to evaluate the potential effect of the administration of two exopolysaccharides (EPS) "EPLB" and "EPB" derived from potential probiotic strains on immune and antioxidant status of European sea bass (Dicentrarchus labrax L.) larvae. In order to find out if the EPSs have an effect on the biochemical composition during the trial period, the nutritional value has been evaluated. The results revealed that expression levels of immune-relevant genes (infg, Il1b, Il8, Il6 and tcr-ß) in the gut and head kidney and the scavenging enzymes (cat, sod, gr) genes in the liver were modulated. In fact, the dietary supplementation with the tested EPSs, significantly enhances the expression of immune-associated genes in the head-kidney, particularly infg and tcrß, as well as catalase gene in liver. During the period of study, EPSs administration did not affect the fatty acid profiles of larvae, which is balanced. This is confirmed by the Docosahexaenoic acid / Eicosapentaenoic acid ratio and demonstrates that EPLB and EPB can be administrated without any negative effect on biochemical composition of European sea bass. The present findings provided evidence that the tested EPSs with antibacterial and antioxidant activities can enhance immune response without negative effect on the biochemical composition. The used EPSs can be considered as a good source of natural functional aquafeed ingredients for European sea bass.

The antioxidant power of horseradish, Armoracia rusticana, underlies antimicrobial and antiradical effects, exerted in vitro.

Armoracia rusticana (AR) was tested for antimicrobial and antioxidants power. The compound demonstrated to inhibit fish pathogens such as Vibrio anguillarum, V. harvey, V. alginolyticus, Aeromonas hydrophila, A. salmonicida, Photobacterium damselae subspecie piscicida, Tenacibaculum marinum and Pseudomonas anguilliseptica,. The total phenolic content and the reducing power resulted higher in the water extract of AR, respect to the hydroalcoolic. In vitro test demonstrated that AR significantly protect cells against death, induced by oxidative stress.