The increasing demand for natural products and biotechnological activities from bees facilitate their widespread use in food preservation and beneficial effects on humans. This study aimed to prepare and characterize the nano-capsules of Qaluiobia (PQG) governorates propolis extracted with water, ethanol and supercritical fluid-carbon dioxide at 50 °C with co-solvent. Propolis bioavailability was analyzed and introduced to prepare crackers to extend their shelf life. Nano-encapsulation was examined using transmission electron microscopy (TEM), differential scanning calorimetry (DSC) and antioxidant activity. Ethanol and supercritical fluid-carbon dioxide (SCF-CO2) at 50 °C with ethanol as co-solvent recorded higher yield, antioxidant activities, total phenolics and total flavonoids. SCF-CO2 extracts had a higher flavonoid concentration. It was revealed that propolis nano-capsules had high-temperature stability and cytotoxic effects against the three tested human cancer cell lines (i.e. PC3, MCF7 and HePG2). The higher overall acceptability of crackers fortified with PQG was achieved with SCF-CO2 at 50 °C and ethanol extract nano-capsules, i.e. 86.57% and 86.29% respectively. The higher ability to retain antioxidant activity reduces the increase of peroxide value (PV), preventing rancidity and increasing the shelf life of crackers during the storage period. Practical application: This study can provide a suitable method for extracting bioactive compounds from propolis, and improve the biological properties and activities by nano-encapsulation, also reveals the extent of its use as a natural antioxidant and anticancer and its application in bakery products as a functional food.

Green microalgae are single-celled eukaryotic organisms that, in recent years, are becoming increasingly important in the nutraceutical, cosmetic, and pharmaceutical fields because of their high content of bioactive compounds. In this study, a particular green microalga was isolated from freshwater highland lakes of Ecuador and morphologically and molecularly identified as Chlamydomonas agloeformis (ChA), and it was studied for nutritional and nutraceutical properties. The phenolic composition and the fatty acids profile of lyophilized cells were determined. The methanolic extract was analyzed for the phenolic compounds profile and the antioxidant capacity by means of in vitro tests. Finally, Human Microvascular Endothelial Cells (HMEC-1) were exploited to explore the capacity of ChA to reduce the endothelial damage induced by oxidized LDL-mediated oxidative stress. The extract showed a good antioxidant ability thanks to the high content in polyphenolic compounds. The observed decrease in HMEC-1 cells endothelial damage also was probably due to the antioxidant compounds present in the extract. Based on the outcomes of our in vitro assays, ChA demonstrated to be a promising source of bioactive compounds possessing exceptional antioxidant capacities which make it a prospective functional food.

Seeds of Vitis vinifera L. with a high content of bioactive compounds are valuable by- products from grape processing. However, little is known about the bioactivity of seeds from other Vitis species. The aim of this study has been to compare the phenolic composition, antimicrobial activ- ity, and antioxidant activity of extracts from seeds of four Vitis species (V. riparia Michx., V. californica Benth., V. amurensis Rupr., and V. vinifera L.). Antioxidant activities were assessed as ferric-reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl radical (DPPHo) scavenging activity, and oxygen radical absorbance capacity (ORAC). The antimicrobial activity was determined using the mi- crodilution method against some Gram-negative (Escherichia coli, Salmonella enterica ser. Typhimurium, and Enterobacter aerogenes) and Gram-positive (Enterococcus faecalis and Staphylococcus aureus) bac- teria. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the phenolic profile of extracts. Flavan-3-ols, procyanidins, phenolic acids, flavonols, anthocyanins, and stilbenoids were detected. (+)-Catechin and (-)-epicatechin turned out to be the most abundant in the phenolic profile of V. amurensis seed extract. Phenolic acids prevailed in the extract from V. vinifera seeds. The V. riparia and V. californica seed extracts had higher contents of most individual phenolics compared to the other Vitis species. They also showed a higher total phenolic content, DPPHo scavenging activity, ORAC, and overall antibacterial activity. Total phenolic content signifi- cantly correlated with antioxidant activity and antimicrobial activity against E. coli. The principal component analysis (PCA) showed discrimination between V. vinifera, V. amurensis, and clustered V. riparia and V. californica with respect to variables. To recapitulate, this research demonstrates that seeds of different Vitis species, especially V. riparia and V. californica, are sources of molecules with antioxidant and antimicrobial activities that can be used in different sectors, such as in the food, cosmetic, and pharmaceutical industries.

Recently, green microalgae have gained importance due to their nutritional and bioactive compounds, which makes them some of the most promising and innovative functional foods. The aim of this study was to evaluate the chemical profile and the in vitro antioxidant, antimicrobial and antimutagenic activity of an aqueous extract of the green microalga Ettlia pseudoalveolaris, obtained from the freshwater lakes of the Ecuadorian Highlands. Human microvascular endothelial cells (HMEC-1) were used to determine the ability of the microalga to reduce the endothelial damage caused by hydrogen peroxide-induced oxidative stress. Furthermore, the eukaryotic system Saccha- romyces cerevisiae was used to evaluate the possible cytotoxic, mutagenic and antimutagenic effect of E. pseudoalveolaris. The extract showed a notable antioxidant capacity and a moderate antibacterial activity mostly due to the high content in polyphenolic compounds. It is likely that the antioxidant compounds present in the extract were also responsible for the observed reduction in endothelial damage of HMEC-1 cells. An antimutagenic effect through a direct antioxidant mechanism was also found. Based on the results of in vitro assays, E. pseudoalveolaris proved to be a good source of bioactive compounds and antioxidant, antibacterial and antimutagenic capacities making it a potential functional food.

In recent years, a wide variety of minimally processed (MP) fruits and vegetables has been worked up together with the development of synthetic additive-free or natural additive-based preservation techniques. As apple is one of the worldwide most widely consumed fruit, the objective of this research was threefold: (i) to verify the effectiveness of an innovative multi-step green process to recover high-add-ed-value antioxidant extracts from organic apple peels. Two different extraction solvents, water or eth-anol at 10%, were used on apple peels;(ii) to evaluate the antimicrobial activity of extracts on MP apples; (iii) to evaluate the ability of the extracts to maintain the physicochemical attributes of fresh cut apple and assess their potentiality as alternative preservative media in comparison with traditional ones, butylat-ed hydroxytoluene (BHT) and citric acid (CA). The extract from 10% ethanol significantly maintained fresh cut apple quality parameters (i.e., browning and texture) similarly to BHT and CA, when compared to untreated fresh cut fruits. Moreover, the apple peel extracts showed an interesting antimicrobial effect to reduce the growth of most relevant pathogenic and deteriorating both positive and negative gram bacteria.

Herein we characterized the bioactive metabolites of the aqueous extract of Kavolì®, a commercial product composed of a mixture of Brassica oleracea leaves, and assessed its potential ameliorating effects in a rat model of non-alcoholic fatty liver disease (NAFLD). Kavolì® extract showed high levels of bioactive compounds and strong in vitro antioxidant activities. Chlorogenic and neochlorogenic acids were identified as the most representative polyphenols. The administration of brassica extract to steatotic rats significantly ameliorated the levels of blood lipids and transaminases, and lipid content and inflammatory markers in liver. Oxidative stress parameters were significantly improved in both liver and brain of steatotic rats. Moreover, plasma and feces levels of short chain fatty acids (SCFAs) were bring back close to control values by Kavolì® treatment, in spite of high fat diet/streptozotocin (HFD/STZ)-induced alterations. The efficacy of Kavolì® in treating hypercholesterolemia, reducing the level of inflammation and cardiovascular disease biomarkers, steatosis and oxidative stress parameters, as well as the ability in modulating SCFAs levels is probably related to the bioactive compounds of the water extract administered to the rat model of NAFLD. In particular, the ameliorating effects are largely attributable to the high content in polyphenols observed in our study.

The aim of this study has been to evaluate the impact of the fermentation process of the spelt from Garfagnana on its chemical composition and short-chain carbohydrates (SCCs) levels, and on the in vitro microbial growth and metabolism. The fermentation process of spelt significantly increases its protein and mannitol content, and decreases its dietary fiber content and fructose, glucose, sucrose, maltose, and raffinose concentration. Fermented spelt modulates the in vitro intestinal microbiota, promoting a stimulation of Lactobacillus and Bifidobacterium spp. growth accompanied by a high production of lactate, acetate, and propionate, both in human gut microbiota from normal weight and obese subjects. The multivariate approach (PCA) combining viable counts and metabolite concentration values has suggested that spelt fermentation could beneficially modulate the gut microbiota from normal weight and obese individuals, stimulating bacteria eliciting anti-inflammatory responses. Further, in vivo studies are recommended for the impact that fermented spelt could have in human nutrition in health and disease.

Many subclinical diseases associated with inflammation occur in sheep during the periparturient period. Clinical symptoms are usually detected at an advanced stage of the disease; therefore, there are considerable risks of permanent health disorders in fetuses or dams. Determination of acute phase proteins (APPs) as markers of inflammation may allow for an earlier diagnosis and effective treatment. Furthermore, multi-fetus pregnancies are often associated with hematological disturbances. The study objective was to compare plasma concentrations of serum amyloid A (SAA), haptoglobin (Hp), fibrinogen (Fb), and cortisol in ewes bearing one and two fetuses in the period from 2 weeks before to 2 weeks after parturition as well as to determine hematological parameters in peripheral blood. There was an important effect of fetal number on APP and cortisol concentrations in periparturient ewes. There was a greater concentration of SAA, Hp, Fb, and cortisol in ewes bearing two fetuses compared with those bearing one fetus. Profiles for APP and cortisol concentrations and hematological parameters were similar for ewes bearing one and two fetuses, and trends were within normal reference ranges for the periparturient period. Furthermore, there were no differences in values for hematological variables between ewes bearing one and two fetuses. with there being no ewes with anemia. In summary, separate determination of the previously undefined physiological ranges of APPs and cortisol for ewes bearing one and two fetuses may facilitate diagnosis of subclinical disorders and enable comparison of laboratory test results with different reference values for ewes bearing different numbers of fetuses.

To preserve quality and extend the shelf life of tomato, a bioactive edible coating was prepared using gelatin wastes from pharmaceutical industry and lyophilised blueberry juice (BJ). The effectiveness of gelatin-coating (G) and G enriched with blueberry juice (GB) was tested, monitoring carotenoids, polyphenols and flavonoids content, the antioxidant activity and the antimicrobial efficiency of coating against the native microflora. After 7 d of storage, coated fruit showed higher phenolic and flavonoids content and increased antioxidant activity, while carotenoids were unaffected by the treatments. The growth of mesophilic bacteria of GB, and the growth of coliform bacteria of G and GB were significantly reduced during the entire period. The results indicate that GB preserved the nutritional quality of tomatoes and that BJ was able to increase the antimicrobial activity of the coating. This paves the way for a possible use of this biodegradable waste polymer as an eco-friendly coating material.

Polyphenol-rich foods could have a pivotal function in the prevention of oxidative stress-based pathologies and antibacterial action. The purpose of this study was to investigate the in vitro antimicrobial activity as well as the in vitro and In Vivo antioxidant capacities of wild Prunus spinosa L. fruit (PSF) from the southeast regions of Italy. The total phenolic content (TPC) was quantified, and the single polyphenols were analyzed by HPLC-DAD, showing high rutin and 4-hydroxybenzoic acid levels, followed by gallic and trans-sinapic acids. PSF extract demonstrated antimicrobial activity against some potentially pathogenic Gram-negative and Gram-positive bacteria. Besides, we investigated the cellular antioxidant activity (CAA) and the hemolysis inhibition of PSF extract on human erythrocytes, evidencing both a good antioxidant power and a marked hemolysis inhibition. Furthermore, an In Vivo experiment with oxidative stress-induced rats treated with a high-fat diet (HFD) and a low dose of streptozotocin (STZ) demonstrated that PSF has a dose-dependent antioxidant capacity both in liver and in brain. In conclusion, the wild Italian Prunus spinosa L. fruit could be considered a potentially useful material for both nutraceutical and food industries because of its antioxidant and antimicrobial effects.

A number of studies have shown the importance of halophyte species as sources of natural antioxidants. Crithmum maritimum is considered an edible and medicinal plant, though in vivo studies of this plant still are necessary to elucidate its potential benefits in alleviating toxicity. The present work was undertaken to investigate the major components and the antioxidant profile of C. maritimum leaf hydro-methanolic extract (CME) and the protective effects of C. maritimum (CM) against CCl4-induced toxicity in rats. Using LC-ESI-MS analysis, 17 phenolic compounds were identified in plant extract, with chlorogenic acid being the major component. The levels of total phenolic acids, total flavonoids and flavonols were ca. 26 mg GAE/g DW, 15.6 mg CE/g DW and 12 mg QE/g DW, respectively. Accordingly, CME showed important in vitro antioxidant activities (DPPH and ORAC). Moreover, CM supplementation to CCl4-intoxicated rats partially restored the impaired hepatic markers (ALT and AST activities and creatinine level), and reduced the CCl4-induced oxidative stress as shown by lipid peroxidation, GSH and protein carbonyl levels. The drug metabolizing system was also evaluated in liver by the measurement of some cytochrome P450-dependent activities such as ECOD, EROD and pNPH and the antioxidant DT-diaphorase, CAT and heme oxygenase activities. Thus, CM treatment reduced the enzymatic perturbations induced by CCl4 and hepatic damages observed from histopathological examination. The obtained results highlight the potential interest of C. maritimum as a source of bioactive compounds with relevant hepatoprotective effects.

Preterm and low birth weight infants require specific nutrition to overcome the accumulated growth deficit, and to prevent morbidities related to postnatal growth failure. In order to guarantee an adequate nutrient-intake, mother's milk, when available, or donated human milk, are usually fortified with additional nutrients, in particular proteins. Fortification with processed ingredients may result in additional intake in oxidative compounds, deriving from extensive heat treatments that are applied during processing. This study aims to compare the in vitro antioxidant activity and oxidative compound content conveyed by different preterm infant foods and fortifiers, namely raw and pasteurized human milk, two different preterm infant formulas, two bovine milk-based fortifiers and two experimental donkey milk-based fortifiers. The antioxidant activity of the infant foods and human milk fortifiers considered in the present research were evaluated as Trolox equivalent antioxidant capacity (TEAC), radical scavenging activity on the radical DPPH and in vitro antioxidant activity using red blood cells as indicators (CAA-RBC). As far as the oxidative content is concerned, the formation of malondialdehydes and protein carbonyls were assessed by HPLC and UV-spectrophotometric method respectively. Univariate and multivariate statistical analyses revealed significant differences between the different products. In particular, the choice of the protein source (hydrolysed vs. whole proteins) may greatly impact the resulting total antioxidant capacity of the preterm meal. The addition of fortifiers to human milk may result in higher antioxidant capacity; among bovine milk proteins, hydrolysed whey proteins resulted to provide higher antioxidant capacity than whole proteins. In general, the use of human milk minimizes the amount of oxidative compound in the diet in comparison to infant formulas, irrespectively of pasteurization or fortification, especially when malondialdehyde content is taken into account. The characterization of macro and micronutrients, bioactive compounds, as well as contaminants, deriving from food alteration, is of primary importance in order to guarantee high quality food products and adequate protein intake, especially when intended for infants, as formulas and fortifiers.

Preterm and low birth weight infants require specific nutrition to overcome the accumulated growth deficit, and to prevent morbidities related to postnatal growth failure. In order to guarantee an adequate nutrient-intake, mother's own milk, when available, or donor human milk, are usually fortified with additional nutrients, in particular proteins. Fortification with processed ingredients may result in additional intake in oxidative compounds, deriving from extensive heat treatments, that are applied during processing. The aim of the present work was to compare the in vitro antioxidant activity and oxidative compound content conveyed by different preterm infant foods and fortifiers, namely raw and pasteurized human milk, two different preterm infant formulas, three bovine milk-based fortifiers and two experimental donkey milk-based fortifiers. Univariate and multivariate statistical analyses revealed significant differences between the different products. The use of human milk minimizes the intake of dietary oxidative compound in comparison to infant formulas, irrespective of pasteurization or fortification, especially as far as malondialdehyde content is concerned. The addition of fortifiers to human milk increases its antioxidant capacity, and the choice of the protein source (hydrolysed vs. whole proteins) differently impacted the resulting total antioxidant capacity of the diet.

Lisosan® Reduction is a plant mixture produced from powder of fermented Triticum aestivum (Lisosan® G), Desmodium adscendens, Malus domestica, Picrorhiza kurroa and Hordeum vulgare. The aim of this study was to characterise the phenolic profile of Lisosan® Reduction and to evaluate the effects of aqueous extract on mice fed a high fat diet (HFD). Syringic acid, trans sinapic acid and neochlorogenic acid were identified by HPLC-DAD to be the dominant polyphenols of Lisosan® Reduction, followed by vitexin, trans p-coumeric acid and trans ferulic acid. Mice treated with aqueous extract of Lisosan® Reduction (60 mg/kg b.w.) showed a significant decrease of serum cholesterol, glucose and triglycerides level and a significant increase of CYP7A1 gene expression, compared to HFD group.

Glucocorticoids (GCs) are currently used for the therapeutic management of cholestatic diseases, but their use and molecular mechanism remain controversial. The aims of this study were 1) to assess the therapeutic effect of a 2-week treatment with the GC dexamethasone on hepatic damage in bile duct-ligated rats; 2) to investigate its effect on the activation of the nuclear receptors (NRs) pregnane X receptor (PXR), constitutive androstane receptor (CAR) and GC receptor (GR), and NF-kappa B, as well as on oxidative stress and bile acid (BA) hepatic composition. Cholestasis was induced by ligation of bile duct (BDL animals) in 16 male Wistar-Kyoto rats, and eight of them were daily treated by oral gavage with 0.125 mg/ml/kg DEX for 14 days. Eight Sham-operated rats were used as controls. Severity of cholestasis was assessed histologically and on plasma biochemical parameters. The nuclear expression of NF-kappa B (p65), GR, PXR and CAR was measured in hepatic tissue by Western Blot. Oxidative stress was evaluated by measuring malondialdehyde, carbonylated proteins, GHS and ROS content in rat livers. LC-MS was used to measure the plasma and liver concentration of 7 BAs. Histological findings and a significant drop in several markers of inflammation (p65 nuclear translocation, mRNA expressions of TNF-alpha, IL-1 beta, IL-6) showed that DEX treatment reversed cholestasis-induced inflammation, and similar results have been obtained with oxidative stress markers. The nuclear expression of p65 and CAR were inversely correlated, with the latter increasing significantly after DEX treatment (p<0.01 vs vehicle). Hepatic BA levels tended to drop in the untreated cholestatic rats, whereas they were similar to those of healthy rats in DEX-treated animals. Plasma BAs decreased significantly in DEX-treated animals with respect to untreated cholestatic rats. In conclusion, DEX reduces inflammation and oxidative stress in BDL rats, and probably CAR is responsible for this effect. Therefore, this NR represents a promising pharmacological target for managing cholestatic and inflammatory liver diseases.

BACKGROUND: Donkey milk is considered as a functional food for sensitive consumers, such as children who are allergic to cow milk.No information is available regarding the effect of farming systems on the quality of donkeymilk. The present study aimed to evaluate the effect of the farming system and lactation stage on donkey milk with respect to gross composition, as well as fat-soluble vitamins and fatty acids (FA). RESULTS: Individual milk samples were collected from lactating jennies (n=53) on the six of the largest farms located in North West Italy. The performance of lactating jennies, herd characteristics and feeding strategies were recorded at each milk sampling. The gross composition of themilk, along with the fat-soluble vitamin content, differed in accordance with the farming system. The lactation stage had limited effects on milk quality. A higher milk fat content corresponded to a higher amount of fresh herbage proportion in the diet, with the highest polyunsaturated fatty acid (PUFA), C18:1c9, C18:3n-3, n-3 FA, retinol and ?-tocopherol content and the lowest concentrations of the FA that are less favorable for human health in the milk of animals fed on only forage diets. CONCLUSION: Extensive farming of dairy donkeys increased the fat content and fat-soluble vitamin concentration of milk and also altered the FA composition to amore favorable profile for human nutrition.

This study evaluated the effects of Tenebrio molitor (TM) larvae meal inclusion in diets for broilers. A total of 160 male broiler chicks (Ross 708) at one-day of age were randomly allotted to four dietary treatments: a control (C) group and three TM groups, in which TM meal was included at 50 (TM5), 100 (TM10), and 150 (TM15) g/kg, respectively. The experimental diets were isonitrogenous and isoenergetic. Each group consisted of five pens as replicates (8 chicks/pen). After the evaluation of growth performance and haematochemical parameters, the animals were slaughtered at 53 days and carcass traits were recorded. Morphometric investigations were performed on duodenum, jejunum, and ileum and histopathological alterations were assessed for liver, spleen, thymus, bursa of Fabricius, kidney, and heart. The live weight (LW) showed a linear (12 and 25 days, P < 0.001 and P < 0.05, maximum with TM15 and TM10) and quadratic (53 days, P < 0.05, maximum with TM5) response to dietary TM meal inclusion. A linear (1 to 12 and 12 to 25 days, P < 0.001, maximum with TM15) and quadratic (12 to 25 days, P = 0.001, maximum with TM15) effect was also observed for the daily feed intake (DFI). The feed conversion ratio (FCR) showed a linear response (25 to 53 and 1 to 53 days, P = 0.001 and P < 0.05, maximum with TM15). Haematological and serum biochemical traits, carcass traits and histopathological findings were not affected by dietary TM meal inclusion (P > 0.05). TM15 birds showed lower villus height (P < 0.05), higher crypt depth (P < 0.05), and lower villus height to crypt depth ratio (P = 0.001) compared with C and TM5. In conclusion, increasing levels of dietary TM meal inclusion in male broiler chickens may improve body weight and feed intake, but negatively affect feed efficiency and intestinal morphology, thus suggesting that low levels may be more suitable. However, no effect on haematochemical parameters, carcass traits, and histological findings were observed in relation to TM meal utilization.

Objective: The present experiment has tested the effect of dietary alfalfa flavonoids (AAF) supplementation on the productive performances, carcass characteristics, meat quality and lipid oxidation of growing rabbits. Methods: One hundred and sixty crossbred rabbits (42 days old) were divided into four groups of forty animals each and were fed either a control diet (AAF0) or an AAF0 diet supplemented with 400, 800, or 1,200 mg of AAF/kg per diet (AAF4, AAF8, and AAF12, respectively) from weaning to slaughtering (102 days old). Performance data were recorded over a period of 60 days. At the end of the trial, 12 rabbits were slaughtered per group, and the carcass characteristics were recorded. Moreover, the plasma, liver and dorsal muscles were sampled from 12 rabbits/group, and were analyzed for lipid oxidation. Results: No significant differences were recorded for the performance, carcass characteristics and meat quality traits except for lightness parameter that was lower in the control group. Dietary AAF supplementation significantly (p<0.01) affected the malondialdehyde (MDA) levels of the frozen meat in a dose-related manner, with the lowest value (0.24 mg MDA/kg fresh meat) recorded in the AAF12 group samples. Conclusion: These findings indicated that the dietary inclusion of AAF in rabbit diets improved muscle oxidation stability with no adverse effects on the growth performance of the animals even if a slight impact on meat lightness color parameter was recorded.