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.

Productive Performance, Egg Quality and Yolk Lipid Oxidation in Laying Hens Fed Diets including Grape Pomace or Grape Extract.

An experiment was conducted to assess in laying hens the effect of including grape pomace (GP, at 30 or 60 g/kg) or grape extract (GE, at 0.5 or 1.0 g/kg) on egg production, feed conversion ratio, protein and polyphenol digestibility, egg weight, egg quality, yolk fatty acid profile and oxidative stability of yolk lipids. No differences were detected among diets for egg production (83.8%, on average) or egg mass (56.8 g/d, on average). However, the average egg weight was lower (p = 0.004) for dietary treatments GP 30, GP 60 and GE 0.5 (67.5 g, on average) than for control hens (68.5 g). Accordingly, in hens fed the GP diets the proportion of XL eggs was lower (p = 0.008) than in control hens, while the proportion of M eggs was higher (p < 0.001) in hens fed the diets GP 30, GP 60 and GE 0.5 than in the control group. The dietary inclusion of both GP and GE decreased daily feed intake (120.9 vs. 125.3 g/d, p < 0.001) and the feed conversion ratio (2.09 vs. 2.18, p = 0.01). Feeding GP at 60 g/kg or GE reduced excreta protein digestibility (54.7 vs. 62.8%, p < 0.001), whereas all GP and GE diets showed higher excreta polyphenol digestibility than the control treatment (57.2 vs. 41.0%, p < 0.001). While yolk colour score was increased with all grape diets (8.12 vs. 7.34, p < 0.001), the dietary inclusion of GP, either at 30 or 60 g/kg, and that of GE at 1.0 g/kg increased the Haugh units of the albumen (80.8 vs. 76.4 Haugh units, p = 0.001). Shell thickness remained unaffected by dietary treatments (365.2 µm, on average). When included in the diet at 60 g/kg, GP reduced the proportion of saturated fatty acids in the yolk (31.6 vs. 32.9%, p = 0.001) and that of monounsaturated fatty acids (39.5 vs. 41.4%, p < 0.001), while it increased the percentage of polyunsaturated fatty acids (28.9 vs. 25.7%, p < 0.001). In fresh eggs, no significant differences were found for the malondialdehyde (MDA) concentration (0.146 mg/kg, on average). In stored eggs, the MDA amount was lower in the eggs of the laying hens fed GP at 60 g/kg than in the eggs of the control hens (1.14 vs. 1.64 mg/kg, p = 0.025). In conclusion, the inclusion of grape pomace, either at 30 or 60 g/kg, and grape extract at 1.0 g/kg in the diet of laying hens improved some egg quality traits, but feeding grape pomace resulted in a lower average weight of eggs. Nevertheless, feeding laying hens with diets containing grape pomace resulted in a higher antioxidant potential in egg yolk than dietary inclusion of grape extract.

Combining Grape Byproducts to Maximise Biological Activity of Polyphenols in Chickens.

Grape seeds (GS) and grape skins (GK) are natural sources of polyphenols with antioxidant capacity. An experiment was conducted to investigate in chickens the effect of including GS and GK (40 g/kg), individually or combined in different proportions (20 g/kg GS-20 g/kg GK; 30 g/kg GS-10 g/kg GK; 10 g/kg GS-30 g/kg GK), in a corn-soybean diet on growth performance, ileal and excreta contents of total extractable polyphenols (TEP) and tannins, ileal digestibility of protein, plasma and meat α-tocopherol concentration and lipid oxidation (assessed by measuring the thiobarbituric acid reactive substances, TBARS) of stored thigh meat. Neither growth performance parameters nor ileal digestibility of protein were affected by dietary treatments. As compared with control birds, chickens fed the grape byproduct diets showed higher ileal (p < 0.001) and excreta (p < 0.001) TEP and tannins contents. Dietary inclusion of grape byproducts increased α-tocopherol concentration both in plasma (p < 0.001) and in thigh meat (p < 0.01 at 1 d; p < 0.001 at 7 d), as compared with the control group. The highest plasma α-tocopherol concentrations were reached with the 30 g/kg GS-10 g/kg GK and 20 g/kg GS-20 g/kg GK combinations. On day 1 of meat storage, no differences on meat α-tocopherol concentration were found among the grape byproducts treatments but on day 7 of storage, the 20 g/kg GS-20 g/kg GK and 10 g/kg GS-30 g/kg GK combinations led to the highest α-tocopherol concentrations in chicken thigh meat. After seven days of refrigerated storage of meat, the TBARS value was lower in chickens fed the grape byproducts diets than in control birds (1.27 vs. 2.49 mg MDA/kg, p < 0.001). Moreover, among the different grape byproduct treatments, the lowest MDA values were reached with the diets containing GK at rates from 20 to 40 g/kg. In conclusion, dietary incorporation of 40 g/kg of GS and GK added separately or combined increased the plasma and meat α-tocopherol content. Furthermore, the combinations of GS and GK with a proportion of GK of at least 50% optimised α-tocopherol concentration both in plasma and in thigh meat and mitigated lipid oxidation in 7-day stored meat.

Short-Term Spirulina (Spirulina platensis) Supplementation and Laying Hen Strain Effects on Eggs' Lipid Profile and Stability.

The aim of the study was to investigate the effect of short-term dietary spirulina supplementation (1% and 3%) and the strain of laying hens (White Leghorn: WL and Rhode Island Red: RIR) on color, nutritional value, and stability of yolk. Egg weight was not affected by any of the studied effects. Yolks from 3%-spirulina supplemented hens had higher retinol and lower α-tocopherol content (p = 0.0001) when compared to control. The supplementation with 1%-spirulina markedly decreased luminosity and increased redness (p = 0.0001) and yellowness (p = 0.0103). Short-term spirulina supplementation slightly modified the fatty acid composition of yolk. The C16-desaturase index increased with the algae, whereas other egg quality indices (hypocholesterolemic, thrombombogenic, n-6/n-3) were not modified. Hen strain mainly affected to the lipid profile. The RIR hens accumulated greater yolk retinol with supplementation doses of 3% (p < 0.05), while the WL hardly suffered changes in the accumulation. Also, yolks from RIR hens had lower C16:0 (p = 0.0001), C18:0 (p = 0.0001), saturated (SAT) (p = 0.0001), and thrombogenic index (p = 0.0001), C20:3n-6 (p = 0.0001), n-6/n-3 ratio (p = 0.003), Δ-6+5-desaturase (p = 0.0005), total elongase indices (p = 0.0001) when compared to WL. Moreover, RIR had higher monounsaturated (MUFA), Δ-9-desaturase and hypocholesterolemic indices (p < 0.05) than WL. A different response to spirulina supplementation was observed for C18:1n-9, MUFA, Δ-9-desaturase and thiesterase indices (p < 0.05) according to hen strain. Yolks from RIR had higher MUFA and Δ-9-desaturase indices than WL at 1%-spirulina supplementation, whereas these parameters were less affected in RIR supplemented with 3%. SAT and Δ-9-desaturase were significantly correlated (r = -0.38 and 0.47, respectively) with retinol content according to a linear adjustment (p < 0.05). Lipid oxidation of yolk was slightly modified by the dietary treatment or hen strain. It was detected a relationship between TBARS and α-tocopherol, C22:5n-3 or C22:6n-3 (p < 0.05). L* and a* were also inversely or positively related with yolk retinol content according to a linear response (p < 0.05). The administration of 1% of spirulina in diets of red hens would be an interesting alternative to get healthier eggs from the nutritional point of view, obtaining an adequate color and without modifications in other yolk quality traits.

Feeding Broiler Chickens with Grape Seed and Skin Meals to Enhance α- and γ-Tocopherol Content and Meat Oxidative Stability.

Grape seeds (GS) and grape skins (GK) are natural sources of polyphenols with featured antioxidant capacity. An experiment was conducted to investigate the effect of these polyphenol sources in diets formulated to contain the same total extractable grape polyphenol content on growth performance, protein and extractable polyphenol digestibility, plasma and meat α- and γ-tocopherol and thigh meat oxidation in broiler chickens. Five experimental diets were formulated: control, control + vitamin E (200 mg/kg), 30 g/kg GS diet, 110 g/kg GK diet, GS + GK diet (a mixture of 24.4 g/kg GS and 13.1 g/kg GK designed to simulate a reconstituted grape pomace). Feeding chickens with 110 g/kg GK reduced (p < 0.001) daily weight gain, worsened (p < 0.001) feed conversion ratio, increased (p < 0.001) non-extractable polyphenol content in the ileum and in the excreta and decreased (p < 0.05) ileal protein digestibility. Regardless of the grape polyphenol source used, the inclusion of grape byproducts in the diets led to an increase of total extractable polyphenol contents in the ileum (p < 0.01) and the excreta (p < 0.001), which resulted (p < 0.001) in a decrease of extractable polyphenol digestibilities. Alpha- and gamma-tocopherol concentrations increased (p < 0.001) in plasma and in seven-day stored meat in birds fed the diet combining GS and GK with respect to the control group. As it happened with the vitamin E supplementation, feeding the combination of GS and GK also reduced (p < 0.001) the concentration of the lipid peroxidation marker (malondialdehyde) in the stored meat of chickens.

Phenolic Metabolites in Plasma and Thigh Meat of Chickens Supplemented with Grape Byproducts.

Grape byproducts are rich sources of polyphenols with powerful antioxidant and health-promoting effects. The impact of supplementing chicken diets with grape byproducts on plasma and thigh meat concentrations of phenolic metabolites was evaluated by analyzing samples by high-performance liquid chromatography quadrupole time of flight mass spectrometry. Chickens were fed three experimental diets: Control diet, Control+8% grape pomace, and Control+0.1% grape seed extract. In plasma, 32 phenolic metabolites were identified, some of which were conjugated catechin/epicatechin metabolites exclusively identified in chickens fed diets enriched in grape byproducts. Also, these chickens showed significantly higher plasmatic concentrations of 21 phenolic metabolites. In thigh meat, 14 phenolic metabolites were identified, but no differences were found between diets. Higher plasmatic tocopherol was found when supplementing diets with grape byproducts, while no changes were observed in meat. Thus, supplementing chicken diets with grape byproducts leads to a significant increase in the circulation of phenolic metabolites and tocopherol.

Impact of a sustained consumption of grape extract on digestion, gut microbial metabolism and intestinal barrier in broiler chickens.

The effect of dietary supplementation with grape extract (GE) at 2.5 and 5.0 g kg-1 of feed on intestinal utilization of polyphenols and gut health of broiler chickens was determined. The ileal digestibility of grape polyphenols was higher for flavan-3-ol monomers [(+)-catechin and (-)-epicatechin] than for dimers (Procyanidins B1 and B2) and galloylated compounds [(-)-epicatechingallate] and no differences among 2.5 and 5.0 g GE per kg dietary treatments were observed. The excreta concentration of benzoic, phenylacetic, phenylpropionic, and cinnamic acids and phenyl-γ-valerolactone phenolic metabolites was higher in birds fed GE, confirming hence the microbial metabolism of grape polyphenols to a relevant extent. Gut morphology and the total ileal mucin content were not modified by the dietary inclusion of GE, but a lower sialic acid concentration was observed in those birds fed a higher concentration of GE. Escherichia coli and lactic-acid bacteria ileal counts were reduced in birds fed GE. Overall, these results prove the extensive intestinal utilization and microbial metabolism of grape polyphenols in broiler chickens. Some antimicrobial and mucin-modulation effects were also observed after a sustained consumption of grape polyphenols during 21 days.