Chlamydomonas agloeformis from the Ecuadorian Highlands: Nutrients and Bioactive Compounds Profiling and In Vitro Antioxidant Activity.

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.

In Vitro Characterization of Antioxidant, Antibacterial and Antimutagenic Activities of the Green Microalga Ettlia pseudoalveolaris.

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 Saccharomyces 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.

Verbascoside-Rich Plant Extracts in Animal Nutrition.

In recent years, the search for dietary intervention with natural products able to sustain animal health and decrease environmental impact, has raised the number of studies pertaining to the use of plants' secondary metabolites. In fact, in livestock, there is a clear relationship between the animals' antioxidant status and the onset of some diseases that negatively affect animal welfare, health, and productive performance. An interesting compound that belongs to the secondary metabolites family of plants, named phenylpropanoids, is verbascoside. The genus Verbascum, which includes more than 233 plant species, is the genus in which this compound was first identified, but it has also been found in other plant extracts. Verbascoside exhibits several properties such as antioxidant, anti-inflammatory, chemopreventive, and neuroprotective properties, that have been evaluated mainly in in vitro studies for human health. The present work reviews the literature on the dietary integration of plant extracts containing verbascoside in livestock. The effects of dietary plant extracts containing verbascoside on the productive performance, antioxidant status, blood parameters, and meat quality in several animal species were evaluated. The present data point out that dietary plant extracts containing verbascoside appear to be a favorable dietary intervention to enhance health, antioxidant status, and product quality in livestock.

Poultry Meat Quality in Antibiotic Free Production Has Improved by Natural Extract Supplement.

Modern consumers are conscious of the relationship between meat quality and animal welfare. Today, slow-growing chickens are associated with a higher broiler welfare. The present work aims to evaluate the effect of dietary natural extract supplementation with polyphenols and seaweed mixtures (PPE) on breast and thigh muscles quality parameters in Hubbard JA57 slow growth chicken in antibiotic-free production. Five hundred Hubbard female chickens (250 from control and 250 from experimental group) were housed on floor pens (10 pens/treatment, 25 birds/pen) and assigned to two experimental groups: a control group (CON) receiving a commercial diet and another group receiving the same diet supplemented with 0.3% of polyphenols and seaweed mixtures (PPE). Dietary supplementation with PPE did not affect (p > 0.05) growth performances. The breast pH tended to be lower (p = 0.062) in PPE groups. The protein content of breast muscles resulted higher in PPE samples (p < 0.05) than controls. The thigh muscles from PPE group showed a lower (p < 0.05) malondialdehyde content than CON during refrigerated storage. In conclusion, the PPE supplement improves breast muscle protein content and oxidative stability of thigh muscle. This feeding practice is suggested to enhance the nutritional and technological parameters of meat Hubbard slow growth chicken in antibiotic-free production.

The hypolipidemic, anti-inflammatory and antioxidant effect of Kavolì® aqueous extract, a mixture of Brassica oleracea leaves, in a rat model of NAFLD.

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.

Long Term Dietary Supplementation with Omega-3 Fatty Acids in Charolais Beef Cattle Reared in Italian Intensive Systems: Nutritional Profile and Fatty Acids Composition of Longissimus lumborum Muscle.

Recently, the quality of beef has received great attention, and health concerns have been focused on fatty acid composition in relation to dietary requirements. The present work aims to evaluate the effect of omega-3 fatty acids (FA) lipid supplement in beef diet on the nutritional characteristics of Longissimus lumborum (LL) muscle. One hundred and eighty Charolais beef were divided in two groups: the control group (CON) received a basal diet and the second one (TR) an isoenergetic diet containing the omega-3 supplement. Dietary treatment did not affect (p > 0.05) growth performances, carcass characteristics and LL colour indices. Cholesterol content resulted lower (p < 0.001) in LL muscle from TR group than CON. The omega-3 FA and conjugated linoleic acid content were higher (p < 0.001) in LL muscle from TR than CON. As expected, LL muscle from TR group showed an increased value of malondialdehyde than CON during refrigerated storage, anyway, remaining within the threshold value of 1 mg/kg meat. In conclusion, the lipid supplement, rich in omega-3 FA improves the fatty acid profile and decreases cholesterol content of LL muscle. This feeding practice is suggested to enhance the nutritional value of meat from beef reared in intensive condition, improving the consumer's health.

Innovative Blood Antioxidant Test in Standardbred Trotter Horses.

In athletic horses, prolonged and intense training gives rise to an imbalance between the production of free radicals and antioxidant molecules, leading to oxidative stress. Considering the relation between exercise and oxidative stress in horses, the present work aims to validate the Kit Radicaux Libres (KRL) test as a tool to verify the influence of taming, training and racing on the total blood antioxidant activity and some haematochemical parameters. Five Italian Standardbred racehorses (two males and three females, aged 12 ± 1 months) from the same training center were selected and monitored upon arrival and during the following year until the racing season. Blood samples were obtained at different timepoints, corresponding to different steps of training. The data showed that KRL values were higher (p < 0.001) before the beginning of the taming period and at 60 days of taming, compared with the training and racing periods; additionally, the total protein value was affected by the training program, whereas no effects of training on muscle enzymes were detected. These results confirm that exercise plays a role in the production of free radicals and show that the KRL test may represent a valid method to determine oxidative stress in athletic horses.

Effects of Lippia citriodora Leaf Extract on Lipid and Oxidative Blood Profile of Volunteers with Hypercholesterolemia: A Preliminary Study.

Lippia citriodora is a plant traditionally used for its anti-inflammatory, antioxidant and antispasmodic effects, as well as for additional biological activities proven in cell culture, animal studies and a small number of human clinical trials. The plant has also shown a marked improvement in blood lipid profile in some animal species. In the present preliminary study, we investigated the effect of a leaf extract on lipid and oxidative blood profile of hypercholesterolemic volunteers. Twelve adults received Lippia citriodora extract caps, containing 23% phenylpropanoids, (100 mg, once a day) for 16 weeks. Selected blood lipids and plasma oxidative markers were measured at baseline and after 4, 8 and 16 weeks of treatment. Compared with baseline, total cholesterol levels significantly decreased and high-density lipoprotein cholesterol increased, while low-density lipoprotein cholesterol and triglycerides showed only a downward trend. Oxidative status was improved due to a decrease in the concentration of total oxidant status, reactive oxygen metabolites and malondialdehyde, and a significant increase in ferric reducing ability of plasma, vitamin A and vitamin E. These preliminary results suggest that dietary supplementation with Lippia citriodora extract can improve the lipid profile, enhance blood antioxidant power, and could be a valuable natural compound for the management of human hypercholesterolemia.

Corneal UV Protective Effects of a Topical Antioxidant Formulation: A Pilot Study on In Vivo Rabbits.

This study aimed to evaluate the protective effect of a topical antioxidant and ultraviolet (UV) shielding action formulation containing riboflavin and D-α-tocopherol polyethylene glycol succinate (TPGS) vitamin E against corneal UV-induced damage in vivo rabbit eyes. In vivo experiments were performed using male albino rabbits, which were divided into four groups. The control group (CG) did not receive any UV irradiation; the first group (IG) was irradiated with a UV-B-UV-A lamp for 30 min; the second (G30) and third (G60) groups received UV irradiation for 30 and 60 min, respectively, and were topically treated with one drop of the antioxidant and shielding formulation every 15 min, starting one hour before irradiation, until the end of UV exposure. The cornea of the IG group showed irregular thickening, detachment of residual fragments of the Descemet membrane, stromal fluid swelling with consequent collagen fiber disorganization and disruption, and inflammation. The cornea of the G30 group showed edema, a mild thickening of the Descemet membrane without fibrillar collagen disruption and focal discoloration, or inflammation. In the G60 group, the cornea showed a more severe thickening, a more abundant fluid accumulation underneath the Descemet membrane with focal detachment, and no signs of severe tissue alterations, as were recorded in the IG group. Our results demonstrate that topical application of eye drops containing riboflavin and TPGS vitamin E counteracts UV corneal injury in exposed rabbits.

Effects of Long-Term Supplementation with Brown Seaweeds and Polyphenols in Rabbit on Meat Quality Parameters.

The objective of the present study was to evaluate the effects of dam and offspring dietary supplementation with a natural feed additive on the growth performance and meat quality parameters of growing rabbits. The growing rabbits are selected from lactating does receiving a control diet (C) or diets supplemented with 0.3% (SP1) and 0.6% (SP2) of feed additive containing brown seaweeds (Laminaria spp.) and plant extracts. In the postweaning phase, the growing rabbits remained in the treatment group defined by their does and the trial lasted 42 days. The average daily feed intake and feed conversion ratio were improved in the rabbit fed 0.6% of the natural feed additive. The cholesterol content tended to be lower in Longissimus lumborum (LL) muscle and decrease in Semimembranosus (SM) muscle (in SP2 -41.36% than controls). The α tocopherol and retinol content were enhanced in both muscles of rabbit fed the natural mixture (SP1 and SP2 groups). An improvement of sensory attributes of texture was observed in both muscles from rabbit fed natural mixture. In conclusion, long term supplementation of both lactating does and offspring with the high dosage of brown seaweed and plant polyphenols improves growth performance and enhances meat nutritional and sensory parameters.

Effects of dietary levels of brown seaweeds and plant polyphenols on growth and meat quality parameters in growing rabbit.

Growth performances, carcass characteristics and meat quality parameters from growing rabbit fed with two levels of dietary brown seaweed (Laminaria spp) and plant polyphenols were investigated. One hundred and forty-four New Zealand White rabbits were allotted into three dietary treatments containing 0 (C), 0.3% (T1), and 0.6% (T2) of brown seaweed and plant polyphenols mixture for 42 days. Growth performances and carcass weight were improved in T1 group. Vitamin A and E content in Longissimus thoracis and lumborum (LTL) and Semimembranosus (SM) muscle were enhanced in the treated groups. In the SM muscle, the oxidative stability was improved in rabbit fed with both dosages of dietary supplement, and the cholesterol content tended to be lower in T1 than in T2 and C groups. The LTL and SM muscle sensory characteristics were improved. In conclusion, dietary integration with a low dosage of brown seaweed and plant polyphenols is a valid strategy for enhance growth performance and produce healthier rabbit meat.

Wild Italian Prunus spinosa L. Fruit Exerts In Vitro Antimicrobial Activity and Protects Against In Vitro and In Vivo Oxidative Stress.

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.

Polyphenolic characterisation of plant mixture (Lisosan® Reduction) and its hypocholesterolaemic effect in high fat diet-fed mice.

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.

Rabbit metabolic response and selected meat quality traits: Evaluation of dietary PLX® 23 and LycoBeads® feed supplement.

Effects of dietary supplementation of verbascoside and lycopene, alone and in combination, on some blood parameters, plasma oxidative status and meat quality traits in intensively reared rabbits were investigated. The test lasted 60 days and was conducted on 200 weaned-rabbits, divided into four groups of 50 animals each (5 animals each cage × 10 repetitions). The control group (CON) received a fattening feed without any feed supplements, whereas the experimental groups received the following: the first, an integration in 22 g of PLX® 23 (Lippia citriodora extract; VB group) per ton of feed; the second, an integration in 100 g of LycoBeads® (Solanum lycopersicum extract; LIC group) per ton of feed; and the third, an integration of both feed supplements in combination at the same doses (LIC+VB group). The dietary supplementation with Lippia citriodora and Solanum lycopersicum resulted in an improvement of the blood lipid profile, oxidative plasma markers, and hepatic and renal activity of treated growing rabbits. Feed additives also produced meat with a lower content in SFA and an increased PUFA content, and inhibited the lipid oxidation improving the oxidative stability of rabbit meat. The dietary supplementation, even reporting no effects on productive parameters and carcass yield, showed an improvement on several blood parameters, as indicator of animal welfare, and on quality and healthy meat markers.

Dietary effect of lemon verbena extract on selected blood parameters and on plasma oxidative profile in Avelignese horses.

The effect of Lippia citriodora extract on selected blood parameters and on plasma oxidative markers in Avelignese horses was evaluated. Twenty-four horses were divided into three groups, consisting of eight animals each. Results of two experimental groups, 0.5 mg of verbascoside per kg of metabolic body weight (bw0.75 ) in the low-dose group (LVB) and 1.0 mg of verbascoside per kg of metabolic body weight (bw0.75 ) in the high-dose group (HVB), were compared to the control group (CON). Groups fed L. citriodora extract (HVB and LVB) showed a significant decrease in triglycerides, total cholesterol and LDL cholesterol (p < .01), bilirubin, and transaminases (p < .05), and an increase in HDL cholesterol (p < .01) compared to the CON group. Oxidative status was improved due to significant decrease in plasma concentration of ROMs and TBARS (p < .01) and increase in levels of vitamin A and vitamin E (p < .01). Based on obtained results, it is assumed that dietary supplementation with L. citriodora extract might find a useful application in horse feeding, with positive impact observed in blood parameters and plasma oxidative markers, with beneficial effects on the physiological welfare of livestock animals.

Cytogenetic tests reveal no toxicity in lymphocytes of rabbit (Oryctolagus cuniculus, 2n=44) feed in presence of verbascoside and/or lycopene.

Phenylpropanoid glycosides (PPG), like other phenolic compounds, are a powerful antioxidants and the Verbascoside (VB) is one of the most active of them. A previous study, by using in vitro exposure of blood human lymphocytes to Verbascoside, reported a significant increasings of chromosome fragility compared to control. In the present study, four homogeneous groups of rabbits were used to test in vivo the VB and/or Lycopene (LP) by feeding the animals without VB and LP (control), in presence of VB or/and LP for 80 days. Lymphocyte cell cultures were performed in three different times: 0, 40 and 80 days of the experiment and the cytogenetic tests that we used [CA-test (Chromosome Abnormalities in terms of chromosome and chromatid breaks) and Sister Chromatid Exchange (SCE-test)] have revealed no mutagenic effects on chromosomes. Indeed, mean values/cell of CA and SCE decreased during the experiment with some difference among and within groups, with significant decreasing value only for some group. The study shows clear evidence that diets rich in Verbascoside (and/or Lycopene) do not originate any mutagenic activity, resulting no cytotoxic for the animals and, suggesting a possible their use in both animal and human diets.

The effects of fermented wheat powder (Lisosan G) on the blood lipids and oxidative status of healthy rabbits.

A number of studies have shown that whole grain products have strong antioxidant effects. Lisosan G, a powder produced from the fermentation of Triticum aestivum, has shown hepatoprotective effects. The aim of this paper was to evaluate the effects of Lisosan G on the serum lipid profile and oxidative status of healthy rabbits. Forty rabbits were fed either a normal diet or a diet containing 10 g, 20 g or 30 g Lisosan G/kg of feed for 60 days. The supplementation of Lisosan G did not cause any alteration of the aspartate aminotransferase (AST), alanine aminotransferase (ALT), bilirubin or creatinine concentrations, but caused a reduction in triglycerides, in low density lipoprotein (LDL) cholesterol, and an increase in high density lipoprotein (HDL) cholesterol. The antioxidant activity of Lisosan G has been confirmed on the basis of the reduction in the blood reactive oxygen metabolites (ROMs) and in the level of the thiobarbituric acid reactive substances (TBARS) together with increased vitamin A and E concentrations in the blood. Moreover, Lisosan G also caused the induction of antioxidant enzymes in the liver and kidney of the treated rabbits. This study has shown the ability of Lisosan G to improve the lipid profile and antioxidant status of the blood in healthy rabbits.

Effect of dietary extruded linseed, verbascoside and vitamin E supplements on yield and quality of milk in Lacaune ewes.

Milk yield and milk qualitative parameters were evaluated in Lacaune ewes on a diet supplemented with extruded linseed, verbascoside and vitamin E. A 98 d-trial was conducted on 44 ewes and started 40±2 d post partum. The animals were divided into four homogeneous groups of eleven animals each; one control group (CON) without extruded linseed and dietary supplements, and the diet of the other three experimental groups was enhanced with extruded linseed (L group), extruded linseed-verbascoside (LVB group), and extruded linseed-verbascoside-vitamin E (LVBE group). All animals individually received an isoenergetic diet, consisting of 700 g concentrated feed and meadow hay ad libitum. Body weight, body condition score, milk yield and milk qualitative parameters were assessed. LVB and LVBE groups resulted in a significant improvement (P<0·05) in milk yield due to the verbascoside supplementation. The extruded linseed supplementation L, LVB and LVBE groups produced a milk fat increase and a better milk fatty acid profile in terms of a higher monounsaturated fatty acid (MUFA), polyunsaturated fatty acid (PUFA) and conjugated linoleic acid (CLA) content and a reduced saturated fatty acid (SFA) content, a lower n-6/n-3 ratio and atherogenic and thrombogenic index. The dietary verbascoside supplementation in the LVB and LVBE group resulted in a better milk quality due to the low cholesterol level and higher vitamin A and E contents, in addition to an increased oxidative stability highlighted by the lower thiobarbituric acid reactive substances (TBARS) level. Thus, the addition of extruded linseed and verbascoside supplements improved milk yield and quality both from a chemical and nutritive point of view.

Corneal epithelial wound healing promoted by verbascoside-based liposomal eyedrops.

Different liposomal formulations were prepared to identify those capable of forming eyedrops for corneal diseases. Liposomes with neutral or slightly positive surface charge interact very well with the cornea. Then these formulations were loaded with verbascoside to heal a burn of corneal epithelium induced by alkali. The cornea surface affected involved in wound was monitored as a function of time. Experimental results were modeled by balance equation between the rate of healing, due to the flow of phenylpropanoid, and growth of the wound. The results indicate a latency time of only three hours and furthermore the corneal epithelium heals in 48 hours. Thus, the topical administration of verbascoside appears to reduce the action time of cells, as verified by histochemical and immunofluorescence assays.