The First Evidence of the Beneficial Effects of Se-Supplementation on In Vitro Cultivated Olive Tree Explants.

Selenium is an essential micronutrient that provides important benefits to plants and humans. At proper concentrations, selenium increases plant growth, pollen vitality, the shelf life of fresh products, and seems to improve stress resistance; these effects can certainly be attributed to its direct and indirect antioxidant capacity. For these reasons, in the present work, the effects of selenium at different dosages on in vitro cultivated olive explants were investigated to observe possible positive effects (in terms of growth and vigor) on the proliferation phase. The work was carried out on four different olive cultivars: "San Felice", "Canino", "Frantoio", and "Moraiolo". The explants were cultured in aseptic conditions on olive medium (OM), with the addition of 4 mg·L-1 of zeatin, 30 g·L-1 of sucrose, and 7 g·L-1 of agar. The experimental scheme included a comparison between explants grown with five different concentrations of Na2SeO4 (0, 10, 20, 40, and 80 mg L-1) added to the medium during three successive subcultures. Interesting information has emerged from the results and all varieties responded to different concentrations of Selenium. The optimal Se dosages varied for each cultivar, but in general, Se concentration between 10 and 40 mg L-1 increased fresh and dry weight of the explants and shoot lengths. Se treatment induced in all cultivars and for all dosages used an increase in total Se content in proliferated explants. Furthermore, as the subcultures proceeded, the ability of the explants to absorb Se did not diminish. The Se content ranged from 8.55 to 114.21 µg kg-1 plant DW in 'Frantoio', from 9.83 to 94.85 µg kg-1 plant DW in 'Moraiolo', from 19.84 to 114.21 µg kg-1 plant DW in 'Canino', and from 20.97 to 95.54 µg kg-1 plant DW in 'San Felice'. In general, the effect of selenium tends to decrease with the progress of subcultures and this suggests a sort of "adaptation" effect of the explants to its presence. The present study highlights for the first time the possibility of using in vitro cultures as biotechnological support to study supplementation with selenium and its effects on in vitro olive plant growth.

Green and simple extraction of free seleno-amino acids from powdered and lyophilized milk samples with natural deep eutectic solvents.

Natural deep eutectic solvents (NADES) were introduced for the extraction of free seleno-amino acids from lyophilized and powdered milk samples. Different NADES were evaluated, and lactic acid:glucose (LGH) showed the highest selenium recoveries. Selenium analysis was performed by inductively coupled plasma mass spectrometry (ICP MS). Se-NADES analysis in ICP MS was optimized according to the radio frequency power and nebulization gas flow rate. Se-NADES extraction was optimized by an experimental design. LGH dilution, LGH volume, sample quantity, and ultrasound time were factors influencing the extraction. Seleno-amino acids were determined by liquid chromatography-ICP MS. After optimization, the limits of detection obtained were 7.37, 8.63, and 9.64 µg kg-1 for selenocysteine, selenomethionine, and seleno-methyl-selenocysteine, respectively. The NADES-extraction is a green procedure with 2 penalty points in the EcoScale. The method was applied to the analysis of powdered milk, lyophilized Se-fortified sheep milk, and ERM-BD151 skimmed milk powder.

Current Knowledge on Selenium Biofortification to Improve the Nutraceutical Profile of Food: A Comprehensive Review.

Selenium (Se) is an important micronutrient for living organisms, since it is involved in several physiological and metabolic processes. Se intake in humans is often low and very seldom excessive, and its bioavailability depends also on its chemical form, with organic Se as the most available after ingestion. The main dietary source of Se for humans is represented by plants, since many species are able to metabolize and accumulate organic Se in edible parts to be consumed directly (leaves, flowers, fruits, seeds, and sprouts) or after processing (oil, wine, etc.). Countless studies have recently investigated the Se biofortification of plants to produce Se-enriched foods and elicit the production of secondary metabolites, which may benefit human health when incorporated into the diet. Moreover, feeding animals Se-rich diets may provide Se-enriched meat. This work reviews the most recent literature on the nutraceutical profile of Se-enriched foods from plant and animal sources.

Effect of Feed Supplemented with Selenium-Enriched Olive Leaves on Plasma Oxidative Status, Mineral Profile, and Leukocyte DNA Damage in Growing Rabbits.

This study investigated the effect of a dietary combination of selenium and olive leaves on rabbit health status in order to evaluate the potential use of these combinations as functional ingredients in feed and food. Sixty weaning rabbits were fed with three diets: control feed (C), control feed + 10% normal olive leaves (OL), or olive leaves enriched in Se (2.17 mg Se/kg d.m.; SeOL). The plasma mineral profile, antioxidant status, and leukocyte DNA damage were determined. Inorganic Se was the most abundant form in the OL diet, while the organic one was higher in SeOL than C and OL. A similar trend was found in the plasma. Protein oxidation showed higher values in both supplemented groups; in addition, dietary Se led to a significant improvement (+ 40%) in ferric reducing ability of plasma (FRAP). A marked reduction in DNA damage (9-fold) was observed in the SeOL group compared to C. The combination of selenium and olive leaves in the diet of growing rabbits increased plasma SeMet and FRAP and reduced leukocyte DNA damage.

Selenium maintains cytosolic Ca2+ homeostasis and preserves germination rates of maize pollen under H2O2-induced oxidative stress.

Selenium (Se) displays antioxidant properties that can be exploited, in plants, to counteract abiotic stresses caused by overly-produced reactive oxygen species (ROS). Here, we show that fertigation of maize crops with sodium selenate effectively protects pollen against oxidative stress. Pollen isolated from Se-treated plants (Se1) and untreated controls (Se0) was incubated in vitro with H2O2 to produce oxidative challenge. Given the impact of ROS on Ca2+ homeostasis and Ca2+-dependent signaling, cytosolic Ca2+ was measured to monitor cellular perturbations. We found that H2O2 disrupted Ca2+ homeostasis in Se0 pollen only, while Se1 samples were preserved. The same trend was observed when Se0 samples were treated with sodium selenate or Se-methionine, which recapitulated in vitro the protective capacity of Se-fertigation. Furthermore, we found that germination rates were much better retained in Se1 as compared to Se0 (46% vs 8%, respectively) after exposure to 20 mM H2O2. The same was observed with Se0 pollen treated with Se-methionine, which is the organic form of Se into which most fertigated sodium selenate converts in the plant. These results, together, show a close correlation between ROS, Ca2+ homeostasis and pollen fertility, and provide strong evidence that Se-fertigation is an excellent approach to preserve or enhance agricultural productivity.

Zea mays L. Grain: Increase in Nutraceutical and Antioxidant Properties Due to Se Fortification in Low and High Water Regimes.

This work aimed to investigate the effect of selenium (Se) and irrigation on the grain yield, on the forms of Se, phenols, and carotenes, and on some antioxidant activities of maize ( Zea mays L.) grains. To reach this goal, a 2 year experiment was undertaken. Maize was fertigated with sodium selenite at the rate of 200 g of Se ha-1 and grown under two water regimes. While the irrigation did not show a clear effect on the selected parameters, Se fertigation increased the contents of inorganic and organic Se forms, xanthophyll, and salicylic acid. Furthermore, while Se fertigation decreased the hydroxycinnamic acid content, generally higher antioxidant activities were found in Se-treated grains than in the control. These findings suggest that Se fertigation increases most of the nutraceutical values of maize grains, which therefore might improve human and livestock health and could increase the maize grain shelf life and its byproducts.

Use of Selenium-enriched olive leaves in the feed of growing rabbits: Effect on oxidative status, mineral profile and Selenium speciation of Longissimus dorsi meat.

In the present study the use of Selenium-fortified olive leaves as potential dietary source of Se in rabbit nutrition was evaluated. Sixty New Zealand White rabbits (35 days of age) were randomly assigned to the following dietary treatments: standard diet (C), and C supplemented with either 10% olive leaves (OL) or 10% Selenium-fortified olive leaves (SeOL; 100 mg/L of foliar spray sodium selenate solution). At 70 days of age, 10 rabbits per group were slaughtered and the oxidative status, mineral profile and Selenium speciation of Longissimus dorsi meat was analyzed. Meat of the SeOL group exhibited better oxidative status (lower TBARS, higher GPx and α-tocopherol values) and a 5-fold higher Se content compared to that of the other treatments. The main Se form was SeMet (7-fold higher in the SeOL group), followed by SeCys2. The present trial demonstrates the possibility of using agro-industrial by-products as ingredients in rabbit feeds, thereby enriching meat bioactive compound content.

The Selenium Supplementation Influences Olive Tree Production and Oil Stability Against Oxidation and Can Alleviate the Water Deficiency Effects.

Foliar fertilization with selenium (Se) may well be beneficial in increasing the nutritional and qualitative values of food in Se-deficient regions such as the Mediterranean Basin, and may contribute to an increase in drought resistance in plants. The present study has considered detachment force, flesh firmness, pigmentation, fresh and dry weight, and oil content of olive drupes from Se fertilized olive orchards (Olea europaea L.) under drought stress and well-watered conditions. This study has also evaluated the total Se, Se amino acid, phenol, carotenoid and chlorophyll contents of EVOO, plus its oxidative stability against oxidation. While there was no change in the ripening indexes and the production of olives generally, Se application did increase the total Se, Se methionine, phenol, and carotenoid and chlorophyll contents. The higher concentration of these (bio) chemical compounds in EVOO obtained from Se fertilized plants might well suggest enhanced antioxidant activity. Consequently, EVOO obtained from Se fertilized trees possesses a higher nutritional value and, as indicated by the greater oxidative stability against oxidation, longer shelf life. In addition, under water deficient conditions, a higher fresh olive weight corresponds to a higher level of phenol, carotenoid and chlorophyll, and the chlorophyll-to-carotenoid ratio in Se fertilized trees would appear to confirm the positive role of selenium in alleviating damage caused by drought stress conditions.

Selenium maintains Ca2+ homeostasis in sheep lymphocytes challenged by oxidative stress.

Selenium (Se) is an essential element in human and animal diets, based upon a widespread range of beneficial effects that are primarily due to its antioxidant properties. While Se can be associated to anti-cancer and anti-diabetic activities, reproductive efficiency, and enhancement of the immune system, the mechanistic details of the corresponding biological processes are still largely elusive. To avoid deficiencies and increase bioavailability, Se it is generally supplied to livestock through Se-supplemented feeds or forage plants fertilized with inorganic Se. While the relationship between Ca2+ and ROS (reactive oxygen species) is well known, only a few studies have addressed the possible involvement of Se in the control of cytosolic Ca2+ in oxidative stress. The results on Ca2+ homeostasis were obtained adding exogenous Se in the form of SeO42- to sheep lymphomonocytes cultured in vitro. In particular, Se strongly attenuated 1mM H2O2-induced alteration of intracellular [Ca2+]C as well as the entry of extracellular Ca2+ into the cells with comparable EC50 values for sodium selenate accounting to 1.72 and 2.28 mM, respectively. In an ex vivo trial, it was observed that Ca2+ homeostasis can effectively be rescued in sheep lymphomonocytes exposed in vivo to a Se concentration of approximately 1.9 mM, that was achieved by feeding sheep with olive leaves previously sprayed with 500 mg/plant Na-selenate. Thus the results obtained suggest that the mode of action of selenium markedly influenced Ca2+-related signaling events. Furthermore, results clearly reveal that the protective effect of Se on Ca2+ homeostasis under oxidative challenge can be clearly and effectively achieved through an appropriate dietary regimen obtained also in a circular economy logic using pruning of olive trees treated to reduce tree drought stress.

Use of olive leaves (whether or not fortified with sodium selenate) in rabbit feeding: Effect on performance, carcass and meat characteristics, and estimated indexes of fatty acid metabolism.

Sixty New Zealand White weaned rabbits were divided into three groups and subjected to different dietary treatments: a standard diet for the control (C), a standard feed supplemented with 10% of plain olive leaves (OL) and a standard feed supplemented with 10% of selenium-fortified olive leaves (100 mg/L of foliar spray sodium selenate solution; SeOL). The productive performance was recorded at the time of slaughter (after 35 days); the carcass and meat traits were determined and estimated indexes of fatty acid metabolism were calculated. No significant differences were found on the rabbit productive performance and the physical-chemical characteristics of the meat. Both group of rabbits on the enriched diet showed leaner and thinner carcasses and a higher meat concentration of oleic acid. The estimated index of Δ5 + Δ6-desaturase, starting from n-6 fatty acids, was lower in both groups supplemented with leaves. The use of selenium-fortified olive leaves, positively affected the lipid oxidative stability of rabbit meat.

Selenium Biofortification in Rice ( Oryza sativa L.) Sprouting: Effects on Se Yield and Nutritional Traits with Focus on Phenolic Acid Profile.

The contents of total Se and of inorganic and organic Se species, as well as the contents of proteins, chlorophylls, carotenoids, and phenolic acids, were measured in 10-day old sprouts of rice ( Oryza sativa L.) obtained with increasing levels (15, 45, 135, and 405 mg Se L-1) of sodium selenite and sodium selenate and with distilled water as control. Increasing Se levels increased organic and inorganic Se contents of sprouts, as well as the content of phenolic acids, especially in their soluble conjugated forms. Moderate levels of sodium selenite (i.e., not higher that 45 mg L-1) appeared the best compromise to obtain high Se and phenolic acid yields together with high proportion of organic Se while limiting residual Se in the germination substrate waste. Se biofortification of rice sprouts appears a feasible and efficient way to promote Se and phenolic acid intake in human diet, with well-known health benefits.

Determination of changes in the concentration and distribution of elements within olive drupes (cv. Leccino) from Se biofortified plants, using laser ablation inductively coupled plasma mass spectrometry.

BACKGROUND: Biofortification of food crops has been used to increase the intake of Se in the human diet, even though this may change the concentration of other elements and modify the nutritional properties of the enriched food. Selenium biofortification programs should include routine assessment of the overall mineral composition of enriched plants. RESULTS: Laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS) was used for the assessment of mineral composition of table olives. Olive trees were fertilized with sodium selenate before flowering. At harvest, the edible parts of drupes proved to be significantly enriched in Se, delivering 6.1 µg g-1 (39% of the RDA for five olives). Such enrichment was followed by significant changes in the concentrations of B, Mg, K, Cr, Mn, Fe and Cu in edible parts, which are discussed for their impact on food quality. CONCLUSION: The biofortification of olive plants has allowed the enrichment of fruits with selenium. Enrichment with selenium has caused an increase in the concentration of other elements, which can change the nutritional quality of the drupes. The analytical technique used well as a valuable tool for routinely determining the chemical composition of all fruit parts. © 2018 Society of Chemical Industry.

Biofortification (Se): Does it increase the content of phenolic compounds in virgin olive oil (VOO)?

Extra-Virgin Olive Oil (EVOO) is a fundamental component of the Mediterranean diet and it may contain several anti-oxidant substances, such as phenols. Previous research has shown that this food may be enriched in phenols by spraying a sodium-selenate solution (100 mg L-1 Se) onto the crop canopy before flowering. The aim of this research was to evaluate the effect of this Se-fertilization before flowering (cv. Leccino) on the phenolic profile of EVOOs, and test to what extent such effects depend on the weather pattern, as observed in two contrasting experimental seasons (2013 and 2014). Results showed that Se-fertilisation enriched EVOOs both in selenium (up to 120 µg kg-1) and in phenols (up to 401 mg kg-1). This latter enrichment was related to an increase in PAL (L-Phenylalanine Ammonia-Lyase) activities and it was largely independent on the climatic pattern. Considering the phenolic profile, oleacein, ligustroside, aglycone and oleocanthal were the most affected compounds and were increased by 57, 50 and 32%, respectively. All these compounds, especially oleacein, have been shown to exert a relevant anti-oxidant activity, contributing both to the shelf-life of EVOOs and to positive effects on human health. It is suggested that Se-fertilisation of olive trees before flowering may be an interesting practice, particularly with poor cultivars and cold and rainy weather patterns, which would normally lead to the production of EVOOs with unfavourable phenolic profile.

Selenium speciation profiles in biofortified sangiovese wine.

Biofortification is an agronomic-based strategy, utilized by farmers, to produce selenium (Se)-enriched food products that may help reduce dietary deficiencies of Se occurring throughout susceptible regions of the world. The foliar exposure route application ensures a high efficiency of Se assimilation by the plant since it does not depend on root-to-shoot translocation. In this study we treated grapevines of Sangiovese variety in the pre-flowering period with sodium selenate (100mg Se L-1). Se content was measured in leaves, fruit at harvest time and in wine respectively in treated and not treated samples with ICP-MS. At harvest, a higher amount of Se in the treated leaves compared to untreated ones was found, 16.0±3.1mgkg-1 dry weight (dw) against 0.17±0.006mgkg-1 dw in the untreated ones. The treated grapes had a content of Se of 0.800±0.08mgkg-1 dw, while that untreated one 0.065±0.025mgkg-1 dw. Immediately after the malolactic fermentation, the wine obtained from treated and untreated vines had a Se content of 0.620±0.09mg Se L-1 and 0.024±0.010mg Se L-1 respectively. In our case the percentage of inorganic Se is 26% of the total Se in the untreated wine, while in Se enriched wine this percentage increase to 47.5% of the total Se. The Se(VI) was the inorganic chemical form more present in enriched wine, probably due to foliar application with selenate. Distributions of Se species suggested being careful to the choice of the enrichment solutions to promote a balanced distribution of different chemical forms, perhaps favouring the accumulation of organic forms.