Enriching Eggs with Bioactive Compounds through the Inclusion of Grape Pomace in Laying Hens Diet: Effect on Internal and External Egg Quality Parameters.

(1) Background: Grapes and their associated by-products (such as grape pomace, GP) stand out for their polyphenol content, which makes them a source of bioactive compounds with antioxidant capacity. The aim of this research was to determine if the inclusion of 50 g/kg of GP in the diet of hens could enrich eggs with antioxidants and to study its effect on internal and external egg quality parameters. (2) Methods: A trial was conducted with two genetic lines of hens, which were fed either a control diet or a diet containing 50 g/kg of GP. Performance, internal and external egg quality, and egg yolk content of vitamins E and A and gallic acid were determined. (3) Results: In eggs laid by hens fed a GP diet, Haugh units and yolk color scores were enhanced, and eggshells became thinner, but without affecting the breaking strength. No dietary effect was observed on the vitamin contents of the yolk. A higher gallic acid content was observed in the yolks of eggs laid by hens fed the GP diet, suggesting that some dietary phenolic compounds could be transferred to the eggs. Hen genetics influenced egg weight, albumen Haugh units, shell thickness, and α- and γ-tocopherol concentration in yolks. (4) Conclusions: Dietary inclusion of GP improved the internal quality of eggs, enriching yolks with a phenolic compound but reducing shell thickness.

Use of grape by-products in aquaculture: New frontiers for a circular economy application.

Grape by-products have already been used in cosmetics, food industries, but also animal feed industry, especially monogastrics and in aquaculture. Grape by-products have been studied for a long time and their principal activities are antimicrobial and antioxidant. Concerning aquaculture, the great demand and necessity to replace animal sources with vegetable ones, has placed grape by-products as possible new phytonutrients with beneficial properties. The purpose of this review is to describe the use of grape by-products in aquaculture, during the last decade, concerning their effects on: 1) gut health and welfare status; 2) growth performances; 3) quality of fillets and flesh during the rearing cycle and shelf-life products. Although other studies highlighted that the high supplementation of grape by-products could negatively affect fish health and growth, due to antinutritional factors (tannins), grape by-products are proven to be valuable phytonutrients that can be incorporated into fish feed to enhance growth and health during rearing conditions. Even in fish products, their utilization has proven to elongate the properties and shelf-life of fillets and minces. Further studies to evaluate the possible integrations or replacements with grape by-products in fish feed in order to evaluate their effectiveness in aquaculture from a sustainable circular economy perspective will be desirable to enhance the use of these products.

Green Extraction Strategy Using Bio-Based Aqueous Biphasic Systems for Polyphenol Valorization from Grape By-Product.

Polyphenols are natural compounds with enhanced antioxidant properties. They are present in relatively high concentrations in fruit/vegetable by-products. Therefore, there is a need for the development of efficient and cost-effective methods for the separation and purification of these valuable compounds. Traditional extraction with organic solvents needs to be switched to novel methods that are more efficient, with reduced extraction times and low consumption of organic solvents. Aiming at developing sustainable processes for the separation and purification of phenolic compounds, we used three model compounds, namely resveratrol, quercetin, and gallic acid, to investigate ionic liquid-based aqueous biphasic systems (IL-ABSs) formed by cholinium-based IL in combination with polypropylene glycol with a molecular mass of 400 g/mol (PPG400). The ABS composition in the two-phase region was selected according to a previously determined phase diagram. Extraction studies indicated the preferential partition of resveratrol and quercetin toward the hydrophobic PPG-rich phase that is mainly dominated by its hydrophobic nature and the strong salting-out effect of ILs. On the other hand, due to its considerably hydrophilic nature, gallic acid preferentially migrates toward the IL phase. The achieved results from grape stem extract demonstrated high extraction efficiencies of cholinium dihydrogen phosphate (~99% for resveratrol for the PPG phase and 78% for gallic acid for the IL phase), with considerable selectivity, demonstrating promising outcomes for potential applications.

Pulsed electric fields-assisted extraction of valuable compounds from red grape pomace: Process optimization using response surface methodology.

Background: The application of Pulsed electric fields as a mild and easily scalable electrotechnology represents an effective approach to selectively intensify the extractability of bioactive compounds from grape pomace, one of the most abundant residues generated during the winemaking process. Objective: This study addressed the optimization of the pulsed electric fields (PEF)-assisted extraction to enhance the extraction yields of bioactive compounds from red grape pomace using response surface methodology (RSM). Methods: The cell disintegration index (Z p ) was identified as response variable to determine the optimal PEF processing conditions in terms of field strength (E = 0.5-5 kV/cm) and energy input (WT = 1-20 kJ/kg). For the solid-liquid extraction (SLE) process the effects of temperature (20-50°C), time (30-300min), and solvent concentration (0-50% ethanol in water) on total phenolic content (TPC), flavonoid content (FC), total anthocyanin content (TAC), tannin content (TC), and antioxidant activity (FRAP) of the extracts from untreated and PEF-treated plant tissues were assessed. The phenolic composition of the obtained extracts was determined via HPLC-PDA. Results: Results demonstrated that the application of PEF at the optimal processing conditions (E = 4.6 kV/cm, WT = 20 kJ/kg) significantly enhanced the permeabilization degree of cell membrane of grape pomace tissues, thus intensifying the subsequent extractability of TPC (15%), FC (60%), TAC (23%), TC (42%), and FRAP values (31%) concerning the control extraction. HPLC-PDA analyses showed that, regardless of the application of PEF, the most abundant phenolic compounds were epicatechin, p-coumaric acid, and peonidin 3-O-glucoside, and no degradation of the specific compounds occurred upon PEF application. Conclusion: The optimization of the PEF-assisted extraction process allowed to significantly enhance the extraction yields of high-value-added compounds from red grape pomace, supporting further investigations of this process at a larger scale.

Grape By-Products as Feedstuff for Pig and Poultry Production.

Grape by-products are exceptional options for replacement of conventional and unsustainable feed sources, since large amounts are generated every year from the winery industry. However, the majority is wasted with severe environmental and economic consequences. The present review aimed to evaluate the effects of grape by-products on pig and poultry growth performance. The most recent literature was reviewed using ScienceDirect and PubMed databases and the results of a total of 16 and 38 papers for pigs and poultry, respectively, were assessed. Fewer studies are documented for pig, but the incorporation of grape by-products up to 9% feed led to an improvement in growth performance with an increase in average daily gain. Conversely, lower levels (<3% feed) are needed to achieve these results in poultry. The beneficial effects of grape by-products on animal performance are mainly due to their antioxidant, antimicrobial, and gut morphology modulator properties, but their high level of cell wall lignification and content of polyphenolic compounds (e.g., tannin) limits nutrient digestion and absorption by monogastric animals. The use of exogenous enzymes or mechanical/chemical processes can provide additional nutritional value to these products by improving nutrient bioavailability. Overall, the valorization of grape by-products is imperative to use them as feed alternatives and intestinal health promoters, thereby contributing to boost circular agricultural economy.

Use of Grape By-Products to Enhance Meat Quality and Nutritional Value in Monogastrics.

Grape by-products could be used in monogastric animals' nutrition to reduce feeding costs with conventional crops (e.g., maize and soybean meal) and to improve meat quality. The main grape by-products with the largest expression worldwide, particularly in the Mediterranean region, are grape pomace, grape seed, grape seed oil and grape skins. These by-products are rich sources of bioactive polyphenols, dietary fiber and polyunsaturated fatty acids (PUFA), more specifically, the beneficial n-3 PUFA, that could be transferred to pork and poultry meat. The potential biological activities, mainly associated with antimicrobial and antioxidant properties, make them putative candidates as feed supplements and/or ingredients capable of enhancing meat quality traits, such as color, lipid oxidation and shelf life. However, grape by-products face several limitations, namely, the high level of lignified cell wall and tannin content, both antinutritional compounds that limit nutrients absorption. Therefore, it is imperative to improve grape by-products' bioavailability, taking advantage of enzyme supplementation or pretreatment processes, to use them as feed alternatives contributing to boost a circular agricultural economy. The present review summarizes the current applications and challenges of using grape by-products from the agro-industrial sector in pig and poultry diets aiming at improving meat quality and nutritional value.

The Effects of Grape Polysaccharides Extracted from Grape By-Products on the Chemical Composition and Sensory Characteristics of White Wines.

There is an increasing interest in the valorization of wine waste by-products. Grape pomace/marc can be an important source of polyphenols but also of polysaccharides (PSs). Therefore, the aim of this work was to extract PSs from grape pomace and musts and incorporate them into wines to improve their quality and valorize these residues. Two white wines were elaborated and treated with four different PS extracts obtained from white grape pomace, white must, a wine purified extract rich in RG-II, and commercial inactivated yeasts. In general, the use of grape PSs extracted from grape pomace or must improve some characteristics of wine, increasing the polysaccharide and volatile concentrations. These PS extracts can be useful to modulate some taste attributes such as an excess of acidity and bitterness and can also prevent the loss of volatile compounds associated with fruity and floral notes over time. This is the first study that shows the effects of grape polysaccharides on the chemical composition and sensory characteristics of white wines. Considering the obtained results, the grape pomace and surplus of musts can be considered valuable sources to obtain polysaccharide-rich products, opening a new opportunity to take advantage of by-products from the wine industry.

Comparison of Antibacterial and Antioxidant Properties of Red (cv. Negramaro) and White (cv. Fiano) Skin Pomace Extracts.

Wine pomace has attracted the attention of the food industry, due to its high content in bioactive compounds, and its multiple healthy activities. In this work, whole and separated skin pomaces from fermented (red) and un-fermented (white) grape by-products were characterized for their antioxidant and antimicrobial activities in order to exploit them as functional food ingredient. Antioxidant activity, measured by both ORAC and TEAC assays, was higher in whole than in skin pomace extracts. The characterization of phenolic composition in whole and skin pomace extracts confirmed the peculiarity of some compounds such as anthocyanins (107.84 + 10.3 mg/g TP) in red skin pomace and a great amount of flavanols (80.73 + 4.04 mg/g TP) in white skin pomace. Whole and skin pomace extracts displayed the same antibacterial activity at 250 µg gallic acid equivalents (GAE)/mL. Red and white skin pomace extracts showed a Minimum Inhibitory Concentration (MIC) of 31.25-62.5 GAE/mL against Staphylococcus aureus and Enterococcus faecalis. Pseudomonas spp. were more sensitive to red skin pomace extracts rather than white skin pomace extracts. Given these results, both red and white pomace extracts could be exploited for future application in food, pharmaceutical and cosmetic industry.

Anthocyanin Recovery from Grape by-Products by Combining Ohmic Heating with Food-Grade Solvents: Phenolic Composition, Antioxidant, and Antimicrobial Properties.

Usually, wine-making by-products are discarded, presenting a significant environmental impact. However, they can be used as a source of bioactive compounds. Moreover, consumers' increasing demand for naturally nutritious and healthy products requires new formulations and food product improvement, together with sustainable, environmentally friendly extraction methods. Thus, this work aimed to compare ohmic heating (OH) with conventional methodology (CONV), using food-grade solvents, mainly water, compared to standard methanol extraction of anthocyanins. No significant differences were found between the CONV and OH for total phenolic compounds, which were 2.84 ± 0.037 and 3.28 ± 0.46 mg/g DW gallic acid equivalent, respectively. The same tendency was found for antioxidant capacity, where CONV and OH presented values of 2.02 ± 0.007 g/100 g and 2.34 ± 0.066 g/100 g ascorbic acid equivalent, respectively. The major anthocyanins identified were malvidin-3-O-acetylglucoside, delphinidin-3-O-glucoside, petunidine-3-O-glucoside, cyanidin-3-O-glucoside, and peonidine-3-O-glucoside. These extracts displayed antimicrobial potential against microorganisms such as Yersinia enterocolitica, Pseudomonas aeruginosa, Salmonella enteritidis, methicillin-sensitive Staphylococcus aureus, a methicillin-resistant Staph. aureus (MRSA), and Bacillus cereus. In conclusion, OH provides similar recovery yields with reduced treatment times, less energy consumption, and no need for organic solvents (green extraction routes). Thus, OH combined with water and citric acid allows a safe anthocyanin extraction from grape by-products, thus avoiding the use of toxic solvents such as methanol, and with high biological potential, including antimicrobial and antioxidant activity.

Enzyme-Assisted Extraction to Obtain Phenolic-Enriched Wine Lees with Enhanced Bioactivity in Hypertensive Rats.

The antihypertensive effect of the soluble fraction of wine lees (WL) from Cabernet variety grapes was recently reported by our group. This blood pressure (BP)-lowering effect was attributed to the presence of flavanols and anthocyanins. In this context, phenolic-enriched wine lees (PWL) could potentially exhibit a stronger bioactivity. Therefore, the aim of this study was to obtain a soluble fraction of WL with increased phenolic content and evaluate its functionality. The PWL were obtained using an enzyme-assisted extraction based on the hydrolysis of WL proteins with Flavourzyme®. They contained 57.20% more total phenolic compounds than WL, with anthocyanins and flavanols being the largest families present. In addition, PWL also showed greater angiotensin-converting enzyme inhibitory and antioxidant activities. Finally, the antihypertensive activity of the PWL was evaluated in spontaneously hypertensive rats. A single dose of 5 mL/kg body weight of PWL showed a greater BP-lowering effect than the one shown by WL. Moreover, this antihypertensive effect was more prolonged than the one produced by the antihypertensive drug Captopril. These results demonstrate that enzymatic protein hydrolysis is a useful method to maximize the extraction of phenolic compounds from WL and to obtain extracts with enhanced functionalities.

Valorization of Winemaking By-Products as a Novel Source of Antibacterial Properties: New Strategies to Fight Antibiotic Resistance.

The emergence of antibiotic-resistance in bacteria has limited the ability to treat bacterial infections, besides increasing their morbidity and mortality at the global scale. The need for alternative solutions to deal with this problem is urgent and has brought about a renewed interest in natural products as sources of potential antimicrobials. The wine industry is responsible for the production of vast amounts of waste and by-products, with associated environmental problems. These residues are rich in bioactive secondary metabolites, especially phenolic compounds. Some phenolics are bacteriostatic/bactericidal against several pathogenic bacteria and may have a synergistic action towards antibiotics, mitigating or reverting bacterial resistance to these drugs. Complex phenolic mixtures, such as those present in winemaking residues (pomace, skins, stalks, leaves, and especially seeds), are even more effective as antimicrobials and could be used in combined therapy, thereby contributing to management of the antibiotic resistance crisis. This review focuses on the potentialities of winemaking by-products, their extracts, and constituents as chemotherapeutic antibacterial agents.

Extraction and purification of high added value compounds from by-products of the winemaking chain using alternative/nonconventional processes/technologies.

Grape byproducts are today considered as a cheap source of valuable compounds since existent technologies allow the recovery of target compounds and their recycling. The goal of the current article is to explore the different recovery stages used by both conventional and alternative techniques and processes. Alternative pre-treatments techniques reviewed are: ultrasounds, pulsed electric fields and high voltage discharges. In addition, nonconventional solvent extraction under high pressure, specifically, supercritical fluid extraction and subcritical water extraction are discussed. Finally alternative purification technologies, for example membrane processing were also examined. The intent is to describe the mechanisms involved by these alternative technologies and to summarize the work done on the improvement of the extraction process of phenolic compounds from winery by-products. With a focus on the developmental stage of each technology, highlighting the research need and challenges to be overcome for an industrial implementation of these unitary operations in the overall extraction process. A critical comparison of conventional and alternative techniques will be reviewed for ethe pre-treatment of raw material, the diffusion of polyphenols and the purification of these high added value compounds. This review intends to give the reader some key answers (costs, advantages, drawbacks) to help in the choice of alternative technologies for extraction purposes.

Natural compounds from grape by-products enhance nutritive value and reduce formation of CML in model muffins.

This study had the objective of determining the effects of the addition of different ingredients and grape by-products (GP) to muffins on CML content. It was found that ingredients, such as salt, baking powder and protein-rich components, reduced CML from 50% to 86%. The use of all ingredients simultaneously caused the highest reduction in CML, suggesting synergistic effects in the muffin formula. Raw cane sugar produced higher amounts of CML than refined sucrose, probably due to metal-ion mediated degradation of fructoselysine. The CML content was correlated with the level of oleic acid at -0.829 and with the level of linoleic acid at 0.913. Muffins enriched with appropriate levels of GP (20%) showed a lowering of the CML level and no significant changes in the sensory profile. GP added to the model system with protein-rich ingredients resulted in the weakest inhibitory effects, probably due to the polyphenol-protein binding mechanism.

Protective effect of grape by-product-fortified breads against cholesterol/cholic acid diet-induced hypercholesterolaemia in rats.

BACKGROUND: New breads fortified with two different forms of grape by-products, namely dried powdered skins (PGP) and freeze-dried extract therefrom (EGP), were characterised and their protective effect against hypercholesterolaemia in rats was studied. RESULTS: The phenolic compound profiles of supplemented breads were dominated by epicatechin and catechin together with appreciable amounts of dimeric procyanidins. Sensory evaluation of enhanced breads revealed that a maximum of 6% PGP or 1.4% EGP could be incorporated to prepare acceptable products. Intake of high-cholesterol/cholic acid diet containing 6% PGP- or 1.4% EGP-fortified bread increased fresh stool weight and significantly reduced protein and fat digestion but did not negatively affect animal growth. PGP- and EGP-fortified breads diminished the negative impact of high-cholesterol/cholic acid diet, lowering total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), lipid peroxidation, glucose and leptin levels, preventing visceral fat accumulation and increasing high-density lipoprotein cholesterol and plasma ferric-reducing antioxidant power levels. Since control bread feeding significantly lowered TC, LDL-C and lipid peroxidation compared with high-fat diet, it may suggested that not only grape by-products but also another components in bread were related to lipid metabolism. CONCLUSION: These results demonstrate that intake of both PGP- and EGP-fortified sourdough mixed rye breads might contribute to a reduction of cardiovascular risk.

Changes in polyphenol and polysaccharide content of grape seed extract and grape pomace after enzymatic treatment.

Grape seed extract and grape pomace are rich sources of polyphenols. The aim of this study was to evaluate the release of polyphenols, the solubilisation of carbohydrate, and the antioxidant capacity of these grape by-products after enzymatic reaction with carbohydrases (cellulolytic and pectinolytic activities) and tannase for 24h. The use of tannase in these by-products, and pectinase in grape pomace changed the galloylated form of catechin to its free form, releasing gallic acid and increasing the antioxidant activity. In grape pomace, cellulase treatment was not efficient for phenolic release and antioxidant activity improvement. The addition of carbohydrases to grape pomace, either alone or in combination, degraded the cell wall polysaccharides, increasing the content of monosaccharides. These results provide relevant data about the potential of pectinase, tannase and combinations of enzymes on the release of polyphenols and monosaccharides from grape by-products, improving the antioxidant capacity and the nutritional value.

Atividade antibacteriana, antioxidante e tanante de subprodutos da uva / Antibacterial, antioxidant and tanning activity of grape by-product

O objetivo neste trabalho foi determinar-se in vitro a atividade antibacteriana, antioxidante e tanante de subprodutos da uva (Vitis vinifera) para seu aproveitamento como ingredientes alternativos na indústria avícola e na conservação de alimentos. Compostos polares foram extraídos da semente de uva desengordurada, em solução contendo acetona, água e ácido acético, resultando em 10,3 por cento de rendimento de extrato de semente de uva desengordurada (ESUD). O ESUD apresentou alta atividade antibacteriana in vitro contra cepas de S. aureus e E. coli, mas não contra cepas de Salmonella sp. A atividade antioxidante do ESUD foi comparável ao ácido ascórbico. A atividade tanante foi considerada baixa para a semente de uva e alta para o ESUD.

The aim of this research was to determine in vitro the antibacterial, antioxidant and tanning activity from grape by-products (Vitis vinifera), for its use as alternative ingredients in the poultry industry and food conservation. Polar compound had been extracted of the defatted grape seed (SUD) in a solution with acetone, water and acetic acid, in the 90: 9.5: 0.5 ratio respectively, resulting in a yield of 10.3 percent of defatted grape seed extract (ESUD). ESUD has high antibacterial activity against strains of S. aureus and E. coli, but not against Salmonella sp. The antioxidant activities of the ESUD had comparable to the ascorbic acid. The capacity to bind proteins was considered low for the seed and high for the ESUD.