Extraction of Bioactive Compounds from the Fruits of Jambolan (Syzygium cumini (L.)) Using Alternative Solvents.

This work demonstrates the effectiveness of using alternative solvents to obtain jambolan extracts with a high content of bioactive compounds compared to conventional organic solvents, being the first study to evaluate the best ecological solvent alternative for Syzygium cumini (L.) Skeels. Five alternative solvents were used for extraction: water at 25 °C (W25), water at 50 °C (W50), water at 75 °C (W75), water with citric acid at 2.4% (CA2), and water with citric acid at 9.6% (CA9) in comparison with three conventional solvents: ethanol (EtOH), water with ethanol at 50% (WE), and water with methanol at 50% (WM). A protocol was then established for the extraction and concentration of samples obtained with these solvents. The highest content of total phenolic compounds (TPCs) in the extracts was obtained with the solvent W75 (1347.27 mg GAE/100 g), while in the concentrates it was the solvents EtOH (3823.03 mg GAE/100 g) and WM (4019.39 mg GAE/100 g). Total monomeric anthocyanins (TMAs) increased by 209.31% and 179.95% in extractions with CA2 and CA9, respectively, compared to pulp (35.57 mg eq c-3-g/100 g), demonstrating that they are the most efficient alternative solvents in this extraction. The levels of bioactive compounds and antioxidant activity varied according to the solvents used. Delphinidin 3,5-diglucoside, cyanidin 3,5-diglucoside, delphinidin 3-glucoside, petunidin 3,5-diglucoside, cyanidin 3-glucoside, peonidin 3,5-diglucoside, malvidin 3,5-diglucoside, petunidin 3-glucoside, and malvidin 3-glucoside were identified in most of the samples by UPLC-MS/MS. This study suggests that a simple procedure using alternative solvents can be used as an environmentally friendly strategy to achieve efficient extraction of bioactive compounds in jambolan.

The Effects of Stocking Density and Food Deprivation on Mucous Cells and Lysozyme Activity in the Skin and Gills of Silver Catfish.

This study aimed to identify the regions of the body surface of silver catfish (Rhamdia quelen) with a higher population of mucous cells in the skin. Additionally, the effects of stressful conditions, such as different stocking densities and food deprivation, on the proliferative response of mucous cells in the skin and gill epithelium and their impact on cutaneous mucous lysozyme activity were investigated. Silver catfish were divided into four experimental groups: high stocking density (32 kg/m3) and fed (HSD-F), high stocking density and fasted (HSD-FS), low stocking density (2.5 kg/m3) and fed (LSD-F), and low stocking density and fasted (LSD-FS). Fish in the fed groups received commercial feed twice a day, amounting to 1% of the tank biomass. After a 14-day experimental period, the fish were anesthetized and euthanized. Samples of cutaneous mucous and skin fragments from seven different points and the second left branchial arch were collected. Histological slides of the skin and gills were stained with PAS + Alcian Blue at pH 2.5, and the epidermal mucous lysozyme activity was assessed using the turbidimetric method. The ventral point in front of the ventral fin was found to be the optimal location for collecting cutaneous epithelia due to its higher density of mucous cells. The population of mucous cells in both the skin and gills varied based on the collection point and treatment applied. The highest lysozyme activity in the epidermal mucous was observed in fish from the HSD-F group. Overall, these findings suggest that stocking density and food deprivation create stressful conditions for silver catfish, which modulate their mucosal response to each situation.

Dietary Supplementation with R-(+)-Limonene Improves Growth, Metabolism, Stress, and Antioxidant Responses of Silver Catfish Uninfected and Infected with Aeromonas hydrophila.

R-(+)-limonene is a monoterpene from plants of the genus Citrus with diverse biological properties. This research evaluated the effects of dietary supplementation with R-(+)-limonene on growth, metabolic parameters in plasma and liver, and the antioxidant and stress responses in silver catfish, Rhamdia quelen, challenged or not with Aeromonas hydrophila. Fish were fed for 67 days with different doses of R-(+)-limonene in the diet (control 0.0, L0.5, L1.0, and L2.0 mL/kg of diet). On the 60th day, a challenge with A. hydrophila was performed. R-(+)-limonene in the diet potentiated the productive performance of the fish. The metabolic and antioxidant responses indicate that R-(+)-limonene did not harm the health of the animals and made them more resistant to the bacterial challenge. Histological findings showed the hepatoprotective effect of dietary R-(+)-limonene against A. hydrophila. Igf1 mRNA levels were upregulated in the liver of fish fed with an L2.0 diet but downregulated with bacterial challenge. The expression levels of crh mRNA were higher in the brains of fish fed with the L2.0 diet. However, the L2.0 diet downregulated crh and hspa12a mRNA expression in the brains of infected fish. In conclusion, the results indicated that R-(+)-limonene can be considered a good dietary supplement for silver catfish.

Chemical Composition, Functional and Anticancer Properties of Carrot.

Plants are a valuable source of drugs for cancer treatment. Daucus carota has been investigated for its health properties. In particular, Daucus carota L. subsp. Sativus, the common edible carrot root, has been found to be rich in bioactive compounds such as carotenoids and dietary fiber and contains many other functional components with significant health-promoting features, while Daucus carota L. subsp. Carrot (Apiacae), also known as wild carrot, has been usually used for gastric ulcer therapy, diabetes, and muscle pain in Lebanon. Here, we review the chemical composition of Daucus carota L. and the functional properties of both edible and wild carrot subspecies. Then, we focus on compounds with anticancer characteristics identified in both Daucus carota subspecies, and we discuss their potential use in the development of novel anticancer therapeutic strategies.

Different Extraction Procedures Revealed the Anti-Proliferation Activity from Vegetable Semi-Purified Sources on Breast Cancer Cell Lines.

Breast cancer (BC) remains the leading cause of mortality in women, despite significant advancements in diagnosis. Thus, the identification of new compounds for its treatment is critical. Phytochemicals are known to exhibit anti-cancer properties. Here, we investigated the anti-proliferation potential of extracts from carrot, Calendula officinalis flower, and Aloe vera on breast cancer vs. epithelial cell lines. Various extraction methods were used, and the proliferative effect of the resulting extracts was assessed by proliferation assay on breast cancer and epithelial cell lines. Carrot, Aloe leaf, and Calendula flower extracts were extracted by hexane and methanol methods, and their semi-purified extracts were able to specifically inhibit the proliferation of breast cancer cell lines. The extract composition was investigated by colorimetric assays, UHPLC-HRMS, and MS/MS analysis. All the extracts contained monogalactosyl-monoacylglycerol (MGMG), while digalactosyl-monoacylglycerol (DGMG) and aloe-emodin were found in Aloe, and glycerophosphocholine (GPC) derivatives were identified in Calendula, except for the isomer 2 detected in carrot, suggesting that their observed different anti-proliferative properties may be associated with the different lipid compounds. Interestingly, Calendula extract was able to strongly inhibit the triple negative breast cancer MDA-MB-231 cell line proliferation (about 20% cell survival), supporting MGMG and GPC derivatives as potential drugs for this BC subtype treatment.

Microencapsulated antioxidant stevia fraction fortifies whey protein and enhances its antidiabetic activity.

Whey protein was fortified with a microencapsulated fraction of Stevia rebaudiana, in the proportion 1:4 (w/w), with maltodextrin from the elite variety of Stevia UEM-13, rich in antioxidant compounds, and evaluated its antioxidant and antidiabetic potential in vitro. The fraction in ethyl acetate, the microencapsulated fraction, the whey protein obtained by membrane and a commercial whey protein were characterized and were also investigated solubility, microencapsulation efficiency and stability and digestion in vitro. In addition, these products and two formulations of the icroencapsulated fraction with the obtained whey protein were tested for their potential to inhibit the α-amylase and α-glucosidase enzyme (antidiabetic activity). The microencapsulated fraction (0.5%) and the supplement fortified with the 20% fraction microencapsulated showed inhibitory potential for the enzyme. As for the α-glucosidase enzyme, all products tested showed inhibition, with the formulation with 1.6% microencapsulated fraction added to whey protein being significantly higher. The microencapsulated fraction showed better solubility and stability, including in vitro digestion analysis, and showed antioxidant and antidiabetic capacity. A sensory evaluation was performed with panelists who regularly consume whey protein supplements and products with stevia and the supplement formulation with 1.6 g microencapsulated stevia per 100 g of whey protein have good sensory acceptance.

Recovery of bioactive compounds from an agro-industrial waste: extraction, microencapsulation, and characterization of jaboticaba(Myrciaria cauliflora Berg) pomace as a source of antioxidant.

This study aimed to evaluate the extraction of bioactive compounds from jaboticaba pomace, produce microcapsules by spray dryer technique, and characterize antioxidant compounds. A factorial experimental design was used in the extraction step. Maltodextrin (DE 10) was used as an encapsulating agent, in a ratio of 1: 1 (w/w), in the microencapsulation process. It was observed the increase of all bioactive compounds analyses comparing jaboticaba pomace with the extract. ATR-FTIR spectroscopy showed a vibrational stretching aromatic ring (1718 - 1731 cm-1) typical for anthocyanins. The Gaussian deconvolution presented extract peak area 7.56% higher than pomace. The encapsulating agent protected anthocyanins during the drying process. Microencapsulation of bioactive compounds from jaboticaba pomace can be useful for food applications whereas they are a rich source of antioxidant compounds. Moreover, the use of agro-industrial waste is promising linked to the use of clean technology as water as an antioxidant extractor.

Molecular insight on the binding of stevia glycosides to bovine serum albumin.

The interaction of the steviol and its glycosides (SG), steviolbioside, and rebaudioside A, with bovine serum albumin (BSA) was studied by absorption and fluorescence spectroscopy techniques alongside molecular docking. The stevia derivatives quenched the fluorescence of BSA by a dynamic quenching mechanism, indicating the interaction between the stevia derivatives and BSA. The binding constant (Kb) of steviol was 100-1000-fold higher than those of SG. The stevia derivative/BSA binding reaction was spontaneous and involved the formation of hydrogen bonds and van der Waals interactions between steviol and steviolbioside with BSA, and water reorganization around the rebaudioside A/BSA complex. Molecular docking pointed out the FA1 and FA9 binding sites of BSA as the probable binding sites of steviol and SG, respectively. In conclusion, steviol enhanced hydrophobicity and small size compared to SG may favor its binding to BSA. As steviol and its glycosides share binding sites on BSA with free fatty acids and drugs, they may be competitively displaced from plasma albumin under various physiological states or disease conditions. These findings are clinically relevant and provide an insight into the pharmacokinetics and pharmacodynamics of the stevia glycosides.

Whey protein enriched with Stevia rebaudiana fraction restores the pancreatic function of streptozotocin induced diabetic rats.

The study aims to analyse the treatment of whey protein enriched with Stevia rebaudiana fraction in insulin secretion and its role mitigating streptozotocin-induced hyperglycemia in rats. Thus, diabetic animals were treated with whey protein enriched with S. rebaudiana fraction or with only the protein isolate or only the Stevia fraction. Insulin level in plasma was measured by radioimmunoassay and the viability of ß cells was detected by immunohistochemistry. The results showed that diabetic animals treated with whey protein enriched with S. rebaudiana fraction had a greater recovery from insulinemia, with plasma levels similar to non-diabetic animals (~ 0.13 ng/mL). In addition, the same group showed a higher number of insulin-positive pancreatic B cells (~ 66%) in immunohistochemistry analysis, while the diabetic groups treated with only the fraction of stevia or whey protein showed 38 and 59% of positive cells, respectively. These results show that the treatment may have restored the viability of streptozotocin-injured pancreatic B cells, and consequently increased insulin secretion, suggesting whey protein enriched with S. rebaudiana fraction can be used an adjunct/supplement in diabetic treatment.

Spray drying encapsulation of stevia extract with maltodextrin and evaluation of the physicochemical and functional properties of produced powders.

This work aimed to formulate and perform physicochemical and functional characterization of maltodextrin microcapsules containing ethanolic extract of stevia, rich in antioxidant compounds, encapsulated by a spray-drying process with two maltodextrins (DE10 and DE19). The powders were named M10 and M19, respectively. We analyzed the physicochemical parameters, antidiabetic activity, cytotoxicity, bioaccessibility of the compounds by in vitro digestion, as well as the structure of the microcapsules by scanning electron microscopy. Microcapsules showed higher solubility (∼35%), lower moisture content (∼29%), and the maltodextrin DE10 had higher efficiency as an encapsulating agent (87%) when compared to DE19 (76%) and showed well-defined spherical structures. The microencapsulation preserved the content of phenolic compounds and antioxidant activity present in the extract (7.2% and 87.5%, respectively). The bioaccessibility of these microencapsulated compounds and antioxidant activity were higher under different conditions of in vitro digestion (mouth, gastric, and intestinal conditions) and showed no cytotoxic effects. We identified 41 compounds (by UHPLC-MS/MS-Qtof) related to the nutritional benefits offered by stevia and the microencapsulation technique can be recommended to preserve bioactive compounds. PRACTICAL APPLICATION: Ethanol extract from stevia leaves contains antioxidant phytochemicals related to the nutritional benefits of stevia. However, this extract presents low solubility and consequently low bioaccessibility under in vitro digestion. The microencapsulation process protects the bioactive compounds of the different pH from digestion and improves the physical-chemical parameters of the extract, increasing its applicability as a possible food additive.

Diphenyl diselenide dietary supplementation protects against fumonisin B1-induced oxidative stress in brains of the silver catfish Rhamdia quelen.

The trend toward using plant-based ingredients in aquafeeds has raised important concerns for aquaculture owing to the negative impacts of mycotoxins on fish health; with emphasis for contamination by fumonisin B1 (FB1). The brain is an important target of FB1; however, study of the pathways linked to brain damage is limited to an analysis of histopathological alterations. Reports have demonstrated the protective effects of dietary supplementation with diphenyl diselenide (Ph2Se2) in the brains of fish subjected to several environmental insults; nevertheless, its neuroprotective effects in fish fed with diets contaminated with FB1 remain unknown. Therefore, the aim of this study was to evaluate whether oxidative damage may be a pathway associated with FB1-induced neurotoxicity, as well as to evaluate whether dietary supplementation with Ph2Se2 prevents or reduces FB1-mediated brain oxidative damage in silver catfish. Brain reactive oxygen species (ROS), lipid peroxidation (LOOH) and protein carbonylation increased on day 30 post-feeding in animals that received FB1-contaminated diets compared to the control group, while brain antioxidant capacity against peroxyl radicals (ACAP) levels and catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) activities were lower. Diphenyl diselenide dietary supplementation avoid increases in brain ROS levels, as well minimizing the augmentation of LOOH levels. Furthermore, Ph2Se2 prevented impairment of brain ACAP levels, as well as GPx and GST activities elicited by FB1-contaminated diets. These data suggest that dietary supplementation with 3 mg/kg Ph2Se2 prevented FB1-induced brain damage in silver catfish, and this protective effect occurred through avoided of excessive ROS production, as well as via prevention of brain lipid damage. Furthermore, Ph2Se2 exerted its neuroprotective effects via ameliorative effects on the enzymatic and non-enzymatic antioxidant defense systems, and may be an approach to prevent FB1-induced brain oxidative stress; however, is not an alternative to prevent the impairment on performance caused by FB1.

Camu-camu bioactive compounds extraction by ecofriendly sequential processes (ultrasound assisted extraction and reverse osmosis).

Technical feasibility of an ecofriendly sequential process (ultrasound assisted extraction and reverse osmosis, or UAE and RO) was evaluated in order to obtain a functional Camu-camu (Myrciaria dubia) product with high vitamin C content. Water was used in the assisted extraction by probe ultrasound (UAE) in an experimental design to evaluate different times, amplitudes and temperatures. The best region for total phenolic (TP) and vitamin C (VC) extraction was 5 min, 60 °C and 30% amplitude. Following extraction, the sample was concentrated by reverse osmosis (R25a, 500 Da, polyamide, and 5 bar area 3 ft2), obtaining a relatively low fouling of 19%. At the end of the sequential process (by HPLC-DAD/UV vis), was obtained a concentrated camu-camu (CC) with high Vitamin C (52.01 ± 0.889 mg/g) and cyanidin-3-glucoside, being respectively 7.0 and 4.5 times higher; also the concentration of phenolic compounds was increased by 3.2 times (25.798 mg GAE/g), and anthocyanins in 6.5 times (66.169 mg of cyanidin-3-glucoside/100 g) as well as high antioxidant activity by all three methods evaluated (increased 3.0, 4.6 and 2.38 times for ABTS, DPPH, FRAP, respectively) by comparing the CC with the initial extract (CS). Twenty compounds were identified by UHPLC-QTOF-MS/MS, highlighting quercetin, gallic acid, p- coumaric, ellagic acid and cyanidin-3-glucoside, and at the first time alnusiin was detected in camu-camu. Therefore, the combination of ultrasound assisted extraction and reverse osmosis can be a promising profitable alternative in order to apply bioactive compounds in food, nutraceuticals and cosmetic matrices, bringing their benefits to consumers.

Diphenyl diselenide modulates splenic purinergic signaling in silver catfish fed diets contaminated with fumonisin B1: An attempt to improve immune and hemostatic responses.

The aim of this study was to determine whether purinergic signaling is a pathway associated with fumonisin B1 (FB1)-induced impairment of immune and hemostatic responses. We also determined whether dietary supplementation with diphenyl diselenide (Ph2Se2) prevents or reduces these effects. Splenic nucleoside triphosphate diphosphohydrolase (NTPDase) activity for adenosine triphosphate (ATP) and adenosine diphosphate (ADP) as substrates and total blood thrombocytes counts were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet, while splenic adenosine deaminase (ADA) activity and metabolites of nitric oxide (NOx) levels were significant higher. Also, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significant lower in silver catfish fed with FB1-contaminated diets than in fish fed with a basal diet. Dietary supplementation with 3 mg Ph2Se2/kg of feed effectively modulated splenic NTPDase (ATP as substrate), ADA, GPx and SOD activities, as well as NOx levels, and was partially effective in the modulation of spleen NTPDase activity (ADP as substrate) and total blood thrombocytes count. These data suggest that splenic purinergic signaling of silver catfish fed with FB1-contaminated diets generates a pro-inflammatory profile that contributes to impairment of immune and inflammatory responses, via reduction of splenic ATP hydrolysis followed possible ATP accumulation in the extracellular environment. Reduction of ADP hydrolysis associated with possible accumulation in the extracellular environment can be a pathophysiological response that restricts the hemorrhagic process elicited by FB1 intoxication. Supplementation with Ph2Se2 effectively modulated splenic enzymes associated with control of extracellular nucleotides (except ADP; that was partially modulated) and nucleosides, thereby limiting inflammatory and hemorrhagic processes.

Whey protein sweetened with Stevia rebaudiana Bertoni (Bert.) increases mitochondrial biogenesis markers in the skeletal muscle of resistance-trained rats.

BACKGROUND: A combination of resistance training and whey protein supplementation is a common practice among athletes and recreational exercisers to enhance muscle growth and strength. Although their safety as food additives is controversial, artificial sweeteners are present in whey protein supplements. Thus, natural sweeteners extracted from the leaves of Stevia rebaudiana are a potential alternative, due to their safety and health benefits. Here, we investigated the effects of whey protein sweetened with S. rebaudiana on physical performance and mitochondrial biogenesis markers in the skeletal muscle of resistance-trained rats. METHODS: Forty male Wistar rats were distributed into four groups: sedentary rats, trained rats, trained rats receiving whey protein and trained rats receiving whey protein sweetened with S. rebaudiana leaf extracts. Resistance training was performed by climbing a ladder 5 days per week, during 8-weeks. The training sessions consisted of four climbs carrying a load of 50, 75, 90, and 100% of the maximum load-carrying capacity which we determined before by performing a maximum load-carrying test for each animal. After this period, we collected plasma and tissues samples to evaluate biochemical, histological and molecular (western blot) parameters in these rats. RESULTS: Dietary supplementation with whey protein sweetened with S. rebaudiana significantly enhanced the maximum load-carrying capacity of resistance-trained rats, compared with non-sweetened whey protein supplementation. This enhanced physical performance was accompanied by an increase in the weight of the gastrocnemius and soleus muscle pads. Although the muscle pad of the biceps brachii was not altered, we observed a significant increase in PGC-1α expression, which was followed by a similar pattern in TFAM protein expression, two important mitochondrial biogenesis markers. In addition, a higher level of AMPK phosphorylation was observed in these resistance-trained rats. Finally, supplementation with whey protein sweetened with S. rebaudiana also induced a significant decrease in retroperitoneal adipocyte diameter and an increase in the weight of brown adipose tissue pads in resistance-trained rats. CONCLUSION: The addition of Stevia rebaudiana leaf extracts to whey protein appears to be a potential strategy for those who want to increase muscular mass and strength and also improve mitochondrial function. This strategy may be useful for both athletes and patients with metabolic disorders, such as obesity and type 2 diabetes.

Dietary addition of rutin impairs inflammatory response and protects muscle of silver catfish (Rhamdia quelen) from apoptosis and oxidative stress in Aeromonas hydrophila-induced infection.

This research aimed to assess the influence of dietary addition of rutin on inflammation, apoptosis and antioxidative responses in muscle of silver catfish (Rhamdia quelen) challenged with Aeromonas hydrophila (A. hydrophila). Fish were split into four groups as follows: control, 0.15% rutin, A. hydrophila, 0.15% rutin + A. hydrophila. After 2 weeks of feeding with standard or rutin diets, fish were challenged or not with A. hydrophila for 1 week. Rutin-added diet abrogates A. hydrophila induced-hemorrhage and inflammatory infiltration. It decreases A. hydrophila induced-apoptosis through decreasing the ratio of Bax to Bcl-2 and increasing phospho-Akt to Akt ratio. It diminishes the A. hydrophila induced-rise in nitric oxide and superoxide anion levels and reestablishes superoxide dismutase activity as well. Although such diet is unable to recover the levels of reduced glutathione (GSH), cysteine and glutamate cysteine ligase, which are depleted as a result of A. hydrophila infection, it diminishes the oxidized glutathione (GSSG) content, thus decreasing GSSG to GSH ratio. It increases the levels of cysteine residues of proteins and diminishes those of thiol-protein mixed disulfides, which were changed after A. hydrophila challenge. Finally, it reduces A. hydrophila induced-lipid peroxidation, markedly elevates ascorbic acid and thus reestablishes total antioxidant capacity, whose levels were decreased after A. hydrophila challenge. In conclusion, the dietary addition of rutin at 0.15% impairs A. hydrophila-induced inflammatory response, inhibits A. hydrophila-induced apoptosis and promotes cell survival. It also reduces the A. hydrophila-induced oxidative stress and stimulates the antioxidative responses in muscle of A. hydrophila-infected silver catfish.

Stevia Nonsweetener Fraction Displays an Insulinotropic Effect Involving Neurotransmission in Pancreatic Islets.

Stevia rebaudiana (Bert.) Bertoni besides being a source of noncaloric sweeteners is also an important source of bioactive molecules. Many plant extracts, mostly obtained with ethyl acetate solvent, are rich in polyphenol compounds that present insulinotropic effects. To investigate whether the nonsweetener fraction, which is rich in phenolic compounds isolated from Stevia rebaudiana with the solvent ethyl acetate (EAF), has an insulinotropic effect, including interference at the terminals of the autonomic nervous system of the pancreatic islets of rats. Pancreatic islets were isolated from Wistar rats and incubated with EAF and inhibitory or stimulatory substances of insulin secretion, including cholinergic and adrenergic agonists and antagonists. EAF potentiates glucose-stimulated insulin secretion (GSIS) only in the presence of high glucose and calcium-dependent concentrations. EAF increased muscarinic insulinotropic effects in pancreatic islets, interfering with the muscarinic receptor subfamily M3. Adrenergic inhibitory effects on GSIS were attenuated in the presence of EAF, which interfered with the adrenergic α2 receptor. Results suggest that EAF isolated from stevia leaves is a potential therapy for treating type 2 diabetes mellitus by stimulating insulin secretion only in high glucose concentrations, enhancing parasympathetic signal transduction and inhibiting sympathetic signal transduction in beta cells.

Effects of dietary microencapsulated Cymbopogon flexuosus essential oil on reproductive-related parameters in male Rhamdia quelen.

In aquaculture, nutrition and supplemented diets have been shown to affect broodstock reproductive performance. In this study, we investigated the effects of dietary supplementation with Cymbopogon flexuosus essential oil (CFEO) microcapsules on reproductive-related parameters in silver catfish (Rhamdia quelen) male broodfish. Adult male broodstocks were separated into three groups according to the concentrations of supplemented CFEO (0.0 = control; 1.0 or 3.0 mL per kg of diet). After 20 days under experimental conditions, the animals were euthanized and the gonads were harvested for gonadosomatic index, sperm analysis, oxidative stress, and histopathology; testosterone levels were measured in the plasma; gene expression of prl, smtl, pomca, and pomcb was assessed in the pituitary gland by real-time PCR. The results showed no alterations on reproductive parameters in R. quelen males treated with Cymbopogon flexuosus essential oil compared to the control-diet animals. In conclusion, CFEO microcapsules supplied for 20 days in the concentrations of 1.00 or 3.00 mL per kilogram of diet did not affect the reproduction criteria evaluated in this study in male silver catfish.

Pretreatment with ethanol as an alternative to improve steviol glycosides extraction and purification from a new variety of stevia.

Leaves of a new variety of Stevia rebaudiana with a high content of rebaudioside A were pretreated with ethanol. The ethanolic extract showed high antioxidant potential and 39 compounds were identified, by UPLC/HRMS, among them one not yet mentioned in the literature for stevia leaves. From the in natura leaves and pretreated leaves, the conditions of aqueous extraction of steviol glycosides were investigated using response surface methodology. The aqueous extracts obtained were purified by ion exchange chromatography techniques and membrane separation methods. The recuperation of steviol glycosides was 4.02g for pretreated leaves and 2.20g for in natura leaves. The level of purity was, respectively, 87% and 84.8%. The results obtained demonstrate that pretreatment increases the yield and purity level of stevia sweeteners by the use of environmentally friendly methodologies and the final product presented acceptable sensory characteristics.

New seminal variety of Stevia rebaudiana: Obtaining fractions with high antioxidant potential of leaves.

The aim of this study was to determine the composition and antioxidant potential of leaves of a new variety of Stevia rebaudiana (Stevia UEM-13). Stevia leaves of UEM-13 contain rebaudioside A as the main glycoside, while most wild Stevia plants contain stevioside. Furthermore can be multiplied by seed, which reduces the cost of plant culture techniques as other clonal varieties are multiplied by buds, requiring sophisticated and expensive seedling production systems. Ethanol and methanol were used in the extraction to determine the bioactive compounds. The methanolic extract was fractionated sequentially with hexane, chloroform, ethyl acetate and isobutanol, and the highest concentration of phenolic compounds and flavonoids was obtained in the ethyl acetate fraction (524.20 mg galic acid equivalent/g; 380.62 µg quercetin equivalent/g). The glycoside content varied greatly among the fractions (0.5% - 65.3%). Higher antioxidant potential was found in the methanol extract and the ethyl acetate fraction with 93.5% and 97.32%, respectively. In addition to being an excellent source for obtaining of extracts rich in glycoside, this new variety can also be used as raw material for the production of extracts or fractions with a significant amount of antioxidant activity and potential to be used as additives in food.

New seminal variety of Stevia rebaudiana: Obtaining fractions with high antioxidant potential of leaves

ABSTRACT The aim of this study was to determine the composition and antioxidant potential of leaves of a new variety of Stevia rebaudiana (Stevia UEM-13). Stevia leaves of UEM-13 contain rebaudioside A as the main glycoside, while most wild Stevia plants contain stevioside. Furthermore can be multiplied by seed, which reduces the cost of plant culture techniques as other clonal varieties are multiplied by buds, requiring sophisticated and expensive seedling production systems. Ethanol and methanol were used in the extraction to determine the bioactive compounds. The methanolic extract was fractionated sequentially with hexane, chloroform, ethyl acetate and isobutanol, and the highest concentration of phenolic compounds and flavonoids was obtained in the ethyl acetate fraction (524.20 mg galic acid equivalent/g; 380.62 µg quercetin equivalent/g). The glycoside content varied greatly among the fractions (0.5% - 65.3%). Higher antioxidant potential was found in the methanol extract and the ethyl acetate fraction with 93.5% and 97.32%, respectively. In addition to being an excellent source for obtaining of extracts rich in glycoside, this new variety can also be used as raw material for the production of extracts or fractions with a significant amount of antioxidant activity and potential to be used as additives in food.