Susana Chamorro

Effects of dietary polyphenol-rich grape products on intestinal microflora and gut morphology in broiler chicks

Grapes have high amounts of phenolic compounds, which can modulate the gut activity as well as modify the structure and function of the gastrointestinal tract. The microbiological activity of avoparcin, grape pomace concentrate, and grape seed extract was evaluated in an in vitro study. An in vivo experiment was also conducted to study the effect of the inclusion of grape pomace concentrate and grape seed extract in the diet of broiler chicks on performance, intestinal microflora (by cultured and terminal restriction fragment length polymorphism methodology), and gut morphology at 21 d of age. Dietary treatments included an antibiotic-free diet (CON), a positive control (AVP; 50 mg/kg of avoparcin), and antibiotic-free diets containing grape pomace concentrate (GPC; 60 g/kg) or grape seed extract (GSE; 7.2 g/kg). Performance was not affected by dietary treatment except in the case of birds fed the GSE diet, which showed decreased weight gain. In the ileal content, birds fed CON and GSE diets had the highest populations of Lactobacillus. Compared with the CON diet, the AVP, GPC, and GSE diets increased the populations of Enterococcus and decreased the counts of Clostridium in the ileal content. In the cecal digesta, birds fed GPC and GSE diets had higher populations of Escherichia coli, Lactobacillus, Enterococcus, and Clostridium than birds in any other treatment group. Animals fed GPC and GSE diets showed a higher biodiversity degree than those fed control diets. The frequency of detection of several potential phenol-degrading bacteria as well as unidentified and uncultured organisms was increased in animals fed GPC and GSE diets. Birds fed the CON diet had longer villi and deeper crypt depth than birds in any other treatment group. The highest villi height:crypt depth ratio corresponded to birds fed GPC and AVP diets and the lowest to those fed CON and GSE diets. In conclusion, dietary polyphenol-rich grape products modify the gut morphology and intestinal microflora and increase the biodiversity degree of intestinal bacteria in broiler chicks.

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.

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.

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.

Effects of dietary grape seed extract on growth performance, amino acid digestibility and plasma lipids and mineral content in broiler chicks.

Polyphenols are chemically and biologically active compounds. Grape seed extracts (GSEs) have been widely used as a human food supplement for health promotion and disease prevention. However, there is little information regarding its application in animal feeds. An experiment was conducted to investigate the effect of inclusion of GSE at 0.025, 0.25, 2.5 and 5.0 g/kg in a wheat soya bean control diet on growth performance, protein and amino acid (AA) digestibility and plasma lipid and mineral concentrations in broiler chickens at 21 days of age. Performance was not affected by dietary treatment except in the case of birds fed the diet with the highest GSE concentration, which showed a worsening of weight gain and feed conversion. Apparent ileal digestibility (AID) of protein was significantly reduced in the birds fed the highest concentration of GSE, which also had a reduction on the AID of arginine, histidine, phenylalanine, cystine, glutamic acid and proline compared with those fed control diet. The inclusion of graded concentration of GSE in the chicken diets caused a significant linear decrease in the concentrations of plasma copper, iron and zinc. Plasma cholesterol, triglycerides and lipoproteins (high-density lipoprotein, low-density lipoprotein and very-low-density lipoprotein) concentrations were not affected by dietary GSE. In conclusion, this study demonstrated that incorporation of GSE in chicken diets up to 2.5 g/kg had no adverse effect on growth performance or protein and AA digestibility. Feed conversion was reduced and growth rate was retarded, when chickens were fed 5 g/kg of GSE. This study also indicated that grape polyphenols reduce the free plasma minerals.

Pollination patterns and plant breeding systems in the Galápagos: 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 archipelagos 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.

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.

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

Alien species can severely disrupt the structure, function and stability of native communities. We evaluated the structure of pollination networks in the three main habitats and in the two seasons on the two most disturbed Galápagos Islands, and investigated how such structure is influenced by invasive plants. Alien plants integrate easily into the communities, but show low impact on overall network structure, except for an increase in network selectiveness. The highly invaded and low diversity humid zone showed the highest nestedness and the lowest modularity. Both pollinators and plants were more generalized during the hot season, when most plants were flowering and networks became more nested.

Addition of exogenous enzymes to diets containing grape pomace: Effects on intestinal utilization of catechins and antioxidant status of chickens

Grape pomace (GP) is a rich source of polyphenols with antioxidant capacity. An experiment was conducted to study the effect of GP phenolic compounds included at 5 and 10%, and the addition (individually or combined) of hydrolyzing enzymes (carbohydrase enzyme complex and tannase at 500ppm) on intestinal utilization of catechins and antioxidant status in broiler chickens. A diet supplemented with 200ppm of α-tocopheryl acetate was also used. Our findings demonstrate the capacity of chickens to digest the monomeric (catechin, epicatechin, gallic acid, and epicatechin-O-gallate) and dimeric (procyanidin B1 and procyanidin B2) catechins present in grape pomace. The addition of enzymes (mainly tannase) hydrolyzed the polymeric structures into smaller catechins, but also promoted a lower digestibility of the monomeric and dimeric catechins suggesting that polymeric structures might favour the intestinal utilization of these catechins. The intestinal accumulation of phenolic compounds generated with tannase and with 10% GP reversed the antimicrobial effect against Clostridium perfringens observed with 5% of GP. Grape pomace improved the antioxidant status of the bird, increasing the α-tocopherol and reducing the iron content on plasma, not affecting the plasma gluthatione. Enzymes modified the intestinal utilization of catechins but not additional protective effect was detected on any of the parameters analyzed to evaluate the antioxidant status.

Effect of added grape seed and skin on chicken thigh patties during chilled storage

The authors thank the MINECO, and CSIC for financial support of Projects AGL2012-31355/GAN and the Intramural 2014470E073. Also we are grateful the CAM and ESI Funds for the financial support the project MEDGAN-CM S2013/ABI2913). Thanks to the MIUR and UNIMOL for the PhD fellowship of Maria Nardoia.