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.

Aqueous Extract of Cocoa Phenolic Compounds Protects Differentiated Neuroblastoma SH-SY5Y Cells from Oxidative Stress.

Cocoa is a rich source of polyphenols, especially flavanols and procyanidin oligomers, with antioxidant properties, providing protection against oxidation and nitration. Cocoa phenolic compounds are usually extracted with methanol/ethanol solvents in order to obtain most of their bioactive compounds; however, aqueous extraction seems more representative of the physiological conditions. In this study, an aqueous extract of cocoa powder has been prepared and chemically characterized, and its potential protective effect against chemically-induced oxidative stress has been tested in differentiated human neuroblastoma SH-SY5Y cells. Neuronal-like cultured cells were pretreated with realistic concentrations of cocoa extract and its major monomeric flavanol component, epicatechin, and then submitted to oxidative stress induced by a potent pro-oxidant. After one hour, production of reactive oxygen species was evaluated by two different methods, flow cytometry and in situ fluorescence by a microplate reader. Simultaneously, reduced glutathione and antioxidant defense enzymes glutathione peroxidase and glutathione reductase were determined and the results used for a comparative analysis of both ROS (reactive oxygen species) methods and to test the chemo-protective effect of the bioactive products on neuronal-like cells. The results of this approach, never tested before, validate both analysis of ROS and indicate that concentrations of an aqueous extract of cocoa phenolics and epicatechin within a physiological range confer a significant protection against oxidative insult to neuronal-like cells in culture.

A Red-Berry Mixture as a Nutraceutical: Detailed Composition and Neuronal Protective Effect.

Recommendations towards increased consumption of fresh fruit and vegetables are well supported by epidemiological and clinical trials. However, in some specific cases, it is difficult to follow these recommendations and the use of nutraceuticals or, in the present work, a freeze-dried fruits mixture can be recommended in order to afford the optimal consumption of dietary polyphenols naturally present in fruits and vegetables. In this work we have carefully characterized a red-berry mixture in terms of polyphenol composition, encountering mainly anthocyanins, which account for a total of 2.8 mg/g as cyanidin-3-glucoside equivalents. Additionally, we have assayed the red-berry blend in a cell model of neurological damage by differentiating the cells and measuring the effect of red-berry polyphenols on cell viability and redox state by flow cytometry. The berry-fruit extract showed an inhibitory effect on differentiated SH-SY5Y ROS formation at a concentration as low as 250 µg/mL (33% inhibition). The results show the potential of this berry-fruit blend for its nutraceutical use in the prevention of the neurodegeneration associated with age or environmental agents.

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.

Sitios anatómicos para la inserción de microtornillos interradiculares como dispositivos de anclaje temporal en tratamientos de ortodoncia / Anatomical sites for the insertion of interradicular miniscrews as temporary anchoring devices in orthodontic treatments

Los microtornillos interradiculares (MTI) son dispositivos de uso temporal diseñados para ser insertados en el hueso alveolar con la finalidad de mejorar el anclaje. El objetivo de este estudio fue presentar una guía para odontólogos que permita identificar los sitios anatómicos más apropiados para la inserción de estos dispositivos, logrando así, una mayor tasa de éxito clínico en los tratamientos. PUBMED, Google Académico y SciELO fueron las bases de datos utilizadas para la búsqueda de la literatura, se consideraron artículos publicados entre los años 2000 y 2020, en inglés y en español. Nuestra revisión sugiere según la bibliografía actual, realizar una planificación al comienzo del tratamiento de ortodoncia, utilizando guías generales para la inserción de MTI a través de mapas visuales de "zonas seguras" que existen entre los espacios interradiculares. Para estos, se consideró que la mínima cantidad de hueso alveolar debía ser de 3 mm.

Interradicular miniscrews (MTI) are temporary devices designed to be inserted into the alveolar bone in order to improve anchorage. The aim of this study was to present a guide for dentists that allows identifying the most appropriate anatomical sites for the insertion of these devices, thus achieving a higher clinical success rate in treatments. PUBMED, Google Scholar and SciELO were the databases used to search the literature; articles published between 2000 and 2020, in English and Spanish, were considered. Our review suggests, according to the current bibliography, planning at the beginning of orthodontic treatment, using general guidelines for the insertion of MTI through visual maps of "safe areas" that exist between the inter-root spaces. For these, it was considered that the minimum amount of alveolar bone should be 3 mm.

Effect of spray drying on the polyphenolic compounds present in purple sweet potato roots: Identification of new cinnamoylquinic acids.

The effect of spray drying on the different polyphenolic compounds present in the root of a purple-fleshed sweet potato variety of Ipomoea batatas native from Peru was performed by HPLC-QTOF-MSMS. Nine anthocyanins, including four peonidin, three cyanidin and two pelargonidin derivatives glycosylated with sophorose and/or glucose and acylated with caffeic, ferulic and p-hydroxybenzoic acid were identified. Twenty nine cinnamoylquinic acids (CiQA), including eight mono-CiQA, fourteen di-CiQA, and five tri-CiQA, were identified on the base of their MS fragmentation profile. Relevant amounts of feruloylquinic acid derivatives were identified. Among them, some di and tri-CiQAs containing feruloyl and caffeoyl moieties in their structures, and di-feruloylquinic acids were reported here, for the first time, in Ipomoea. Spray drying process negatively affected the different phenolic groups, with polyphenol losses representing around 90% of the initial amounts. Mono-CiQAs presenting feruloyl moieties and mono acylated peonidin derivatives with p-hydroxybenzoic acid were the less affected compounds.

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.

Response of epilithic diatom communities to environmental gradients along an Ecuadorian Andean River.

Epilithic diatom communities are particularly suitable for the evaluation of freshwater quality. In Ecuador, however, no water quality index includes this biotic parameter. This work is the first attempt in the country to determine the composition of epilithic diatom communities associated with different degrees of eutrophication. This was accomplished by measuring physical, chemical and microbiological variables at five sampling sites along the Pita River, Ecuador, from August to December 2016. The results indicate a clear gradient of eutrophication from sampling sites P1 (good water) in the upper reaches to P5 (bad water) located in the lower reaches. Concerning diatom analyses, the results indicated a high diversity for tropical areas in terms of species richness, varying from S=34 in headwaters to S=42 downstream. Moreover, the results obtained suggest a lack of concordance with the trophic values given to some of the epilithic diatoms in the literature. There were also species that seem to be sensitive to downstream nutrient increases that were not considered as bioindicators in previous studies. We concluded that the trophic values of diatom species available in the scientific literature are not directly applicable to their sites in the Pita River. Hence, it is necessary to establish a trophic diatom index for the Andean region of Ecuador.

Space, time and aliens: charting the dynamic structure of Galápagos pollination networks.

Oceanic archipelagos are threatened by the introduction of alien species which can severely disrupt the structure, function and stability of native communities. Here we investigated the pollination interactions in the two most disturbed Galápagos Islands, comparing the three main habitats and the two seasons, and assessing the impacts of alien plant invasions on network structure. We found that the pollination network structure was rather consistent between the two islands, but differed across habitats and seasons. Overall, the arid zone had the largest networks and highest species generalization levels whereas either the transition between habitats or the humid habitat showed lower values. Our data suggest that alien plants integrate easily into the communities, but with low impact on overall network structure, except for an increase in network selectiveness. The humid zone showed the highest nestedness and the lowest modularity, which might be explained by the low species diversity and the higher incidence of alien plants in this habitat. Both pollinators and plants were also more generalized in the hot season, when networks showed to be more nested. Alien species (both plants and pollinators) represented a high fraction (∼56 %) of the total number of interactions in the networks. It is thus likely that, in spite of the overall weak effect we found of alien plant invasion on pollination network structure, these introduced species influence the reproductive success of native ones, and by doing so, they affect the functioning of the community. This certainly deserves further investigation.

Safety assessment of chronic oral exposure to iron oxide nanoparticles.

Iron oxide nanoparticles with engineered physical and biochemical properties are finding a rapidly increasing number of biomedical applications. However, a wide variety of safety concerns, especially those related to oral exposure, still need to be addressed for iron oxide nanoparticles in order to reach clinical practice. Here, we report on the effects of chronic oral exposure to low doses of γ-Fe2O3 nanoparticles in growing chickens. Animal observation, weight, and diet intake reveal no adverse signs, symptoms, or mortality. No nanoparticle accumulation was observed in liver, spleen, and duodenum, with feces as the main excretion route. Liver iron level and duodenal villi morphology reflect the bioavailability of the iron released from the partial transformation of γ-Fe2O3 nanoparticles in the acid gastric environment. Duodenal gene expression studies related to the absorption of iron from γ-Fe2O3 nanoparticles indicate the enhancement of a ferric over ferrous pathway supporting the role of mucins. Our findings reveal that oral administration of iron oxide nanoparticles is a safe route for drug delivery at low nanoparticle doses.

Invaders of pollination networks in the Galapagos Islands: emergence of novel communities.

The unique biodiversity of most oceanic archipelagos is currently threatened by the introduction of alien species that can displace native biota, disrupt native ecological interactions, and profoundly affect community structure and stability. We investigated the threat of aliens on pollination networks in the species-rich lowlands of five Galápagos Islands. Twenty per cent of all species (60 plants and 220 pollinators) in the pooled network were aliens, being involved in 38 per cent of the interactions. Most aliens were insects, especially dipterans (36%), hymenopterans (30%) and lepidopterans (14%). These alien insects had more links than either endemic pollinators or non-endemic natives, some even acting as island hubs. Aliens linked mostly to generalized species, increasing nestedness and thus network stability. Moreover, they infiltrated all seven connected modules (determined by geographical and phylogenetic constraints) of the overall network, representing around 30 per cent of species in two of them. An astonishingly high proportion (38%) of connectors, which enhance network cohesiveness, was also alien. Results indicate that the structure of these emergent novel communities might become more resistant to certain type of disturbances (e.g. species loss), while being more vulnerable to others (e.g. spread of a disease). Such notable changes in network structure as invasions progress are expected to have important consequences for native biodiversity maintenance.

Pollination patterns and plant breeding systems in the Galapagos: a review.

BACKGROUND: Despite the importance of the Galápagos Islands for the development of central concepts in ecology and evolution, the understanding of many ecological processes in this archipelago is still very basic. One such process is pollination, which provides an important service to both plants and their pollinators. The rather modest level of knowledge on this subject has so far limited our predictive power on the consequences of the increasing threat of introduced plants and pollinators to this unique archipelago. SCOPE: As a first step toward building a unified view of the state of pollination in the Galápagos, a thorough literature search was conducted on the breeding systems of the archipelago's flora and compiled all documented flower-visitor interactions. Based on 38 studies from the last 100 years, we retrieved 329 unique interactions between 123 flowering plant species (50 endemics, 39 non-endemic natives, 26 introduced and eight of unknown origin) from 41 families and 120 animal species from 13 orders. We discuss the emergent patterns and identify promising research avenues in the field. CONCLUSIONS: Although breeding systems are known for <20 % of the flora, most species in our database were self-compatible. Moreover, the incidence of autogamy among endemics, non-endemic natives and alien species did not differ significantly, being high in all groups, which suggests that a poor pollinator fauna does not represent a constraint to the integration of new plant species into the native communities. Most interactions detected (approx. 90 %) come from a single island (most of them from Santa Cruz). Hymenopterans (mainly the endemic carpenter bee Xylocopa darwinii and ants), followed by lepidopterans, were the most important flower visitors. Dipterans were much more important flower visitors in the humid zone than in the dry zone. Bird and lizard pollination has been occasionally reported in the dry zone. Strong biases were detected in the sampling effort dedicated to different islands, time of day, focal plants and functional groups of visitors. Thus, the existing patterns need to be confronted with new and less biased data. The implementation of a community-level approach could greatly increase our understanding of pollination on the islands and our ability to predict the consequences of plant invasions for the natural ecosystems of the Galápagos.

Lactase persistence, NOD2 status and Mycobacterium avium subsp. paratuberculosis infection associations to Inflammatory Bowel Disease.

BACKGROUND: Inflammatory Bowel Disease (IBD), which includes both Crohn's disease (CD) and ulcerative colitis (UC), is caused by a complex interplay involving genetic predisposition, environmental factors and an infectious agent. Mycobacterium avium subsp. paratuberculosis (MAP) is a promising pathogen candidate since it produces a chronic intestinal inflammatory disease in ruminants that resembles CD in humans. MAP is a ubiquitous microorganism, although its presence in the food chain, especially in milk from infected animals, is what made us think that there could be an association between lactase persistence (LP) and IBD. The LCT mutation has brought adaptation to dairy farming which in turn would have increased exposure of the population to infection by MAP. NOD2 gene mutations are highly associated to CD. METHODS: In our study, CD and UC patients and controls from the North of Spain were genotyped for the lactase gene (LCT) and for three NOD-2 variants, R702W, G908R and Cins1007fs. MAP PCR was carried out in order to assess MAP infection status and these results were correlated with LCT and NOD2 genotypes. RESULTS: As for LP, no association was found with IBD, although UC patients were less likely to present the T/T-13910 variant compared to controls, showing a higher C-allele frequency and a tendency to lactase non-persistence (LNP). NOD2 mutations were associated to CD being the per-allele risk higher for the Cins1007fs variant. MAP infection was more extended among the healthy controls (45.2%) compared to CD patients (21.38%) and UC patients (19.04%) and this was attributed to therapy. The Asturian CD cohort presented higher levels of MAP prevalence (38.6%) compared to the Basque CD cohort (15.5%), differences also attributed to therapy. No interaction was found between MAP infection and LCT or NOD2 status. CONCLUSIONS: We conclude that LP is not significantly associated with IBD, but that MAP infection and NOD2 do show not mutually interacting associations with IBD.

An extensive alien plant inventory from the inhabited areas of galapagos.

BACKGROUND: Plant invasions are causing habitat degradation in Galapagos. Problems are concentrated on the four inhabited islands. Plants introduced to rural areas in the humid highlands and urban areas on the arid coast act as foci for invasion of the surrounding Galapagos National Park. METHODOLOGY/PRINCIPAL FINDINGS: Here we present results of the most comprehensive inventory to date of alien vascular plants in the inhabited areas of Galapagos. The survey was conducted between 2002 and 2007, in 6031 properties (97% of the total) on Floreana, Isabela, San Cristobal and Santa Cruz Islands. In total 754 alien vascular plant taxa were recorded, representing 468 genera in 123 families. Dicotyledons represented 554 taxa, monocotyledons 183, there were 7 gymnosperms and 10 pteridophytes. Almost half (363) of the taxa were herbaceous. The most represented families were Fabaceae (sensu lato), Asteraceae and Poaceae. The three most recorded species in the humid rural areas were Psidium guajava, Passiflora edulis and Bryophyllum pinnatum, and in the dry urban areas, Aloe vera, Portulaca oleracea and Carica papaya. In total, 264 (35%) taxa were recorded as naturalized. The most common use for taxa was ornamental (52%). CONCLUSIONS/SIGNIFICANCE: This extensive survey has increased the known alien vascular flora of Galapagos by 257 species, giving a ratio of alien to native taxa of 1.57:1. It provides a crucial baseline for plant invasion management in the archipelago and contributes data for meta analyses of invasion processes worldwide. A repeat of the survey in the future would act as an effective early detection tool to help avoid further invasion of the Galapagos National Park.