Search

ERANET grant BBSRC BB/M028291/1, Swedish Research Council Formas 2014-01840, ARN (Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria) 291815, European Cooperation in Science and Technology (COST) CA15224, UK Research & Innovation (UKRI) Biotechnology and Biological Sciences Research Council (BBSRC) BB/P013759/1

Broiler chickens have an extreme physiology (rapid growth rates) that challenges the correct bone mineralization, being an interesting animal model for studying the development of bone pathologies. This work studies in detail how the mineralization, chemistry, and structural organization of tibiae bone in broiler chickens change with age during the first 5 wk (37 D) from hatching until acquiring the final weight for slaughter. During the early growth phase (first 2 wk), the rapid addition of bone tissue does not allow for bone organic matrix to fully mineralize and mature, and seems to be a critical period for bone development at which bone mineralization cannot keep pace with the rapid growth of bones. The low degree of bone mineralization and large porosity of cortical bone at this period might be responsible of leg deformation and/or other skeletal abnormalities commonly observed in these birds. Later, cortical bone porosity gradually decreases and the cortical bone became fully mineralized (65%) at 37 D of age. At the same time, bone mineral acquires the composition of mature bone tissue (decreased amount of carbonate, higher crystallinity, Ca/P = 1.68). However, the mineral part was still poorly organized even at 37 D. The oriented fraction was about 0.45 which means that more than half of apatite crystals within the mineral are randomly oriented. Mineral organization (crystal orientation) had an important contribution to bone-breaking strength. Nevertheless, locally determined (at tibia mid-shaft) bone properties (i.e., cortical thickness, crystal orientation) has only a moderate correlation (R2 = 0.33) with bone breaking strength probably due to large and highly heterogeneous porosity of bone that acts as structural defects. On the other hand, the total amount of mineral (a global property) measured by total ash content was the best predictor for breaking strength (R2 = 0.49). Knowledge acquired in this study could help in designing strategies to improve bone quality and reduce the incidence of skeletal problems in broiler chickens that have important welfare and economic implications.

Osteoporosis and bone fractures are a severe problem for the welfare of laying hens, with genetics and environment, such as housing system, each making substantial contributions to bone strength. In this work, we performed genetic analyses of bone strength, bone mineral density, and bone composition, as well as body weight, in 860 commercial crossbred laying hens from 2 different companies, kept in either furnished cages or floor pens. We compared bone traits between housing systems and crossbreds and performed a genome-wide association study of bone properties and body weight. As expected, the 2 housing systems produced a large difference in bone strength, with layers housed in floor pens having stronger bones. These differences were accompanied by differences in bone geometry, mineralization, and chemical composition. Genome scans either combining or independently analyzing the 2 housing systems revealed no genome-wide significant loci for bone breaking strength. We detected 3 loci for body weight that were shared between the housing systems on chromosomes 4, 6, and 27 (either genome-wide significant or suggestive) and these coincide with associations for bone length. In summary, we found substantial differences in bone strength, content, and composition between hens kept in floor pens and furnished cages that could be attributed to greater physical activity in pen housing. We found little evidence for large-effect loci for bone strength in commercial crossbred hens, consistent with a highly polygenic architecture for bone strength in the production environment. The lack of consistent genetic associations between housing systems in combination with the differences in bone phenotypes could be due to gene-by-environment interactions with housing system or a lack of power to detect shared associations for bone strength.

(1) Background: Nowadays the industry aims to improve lay persistency for extended cycles (100 weeks or longer) to make egg production more sustainable. However, intensive egg pro‐ duction challenges hen health, inducing severe osteoporosis and the incidence of bone fractures. In this study, the relationship between bone quality and egg production, and/or eggshell quality, was evaluated at the end of an extended laying cycle of 100 weeks, comparing groups of hens with dif‐ ferent production and eggshell quality parameters; (2) Methods: Quality parameters of egg (as weight, egg white height), eggshell (as thickness, weight, breaking strength, elasticity and micro‐ structure) and tibiae bone (weight, diameter, cortical thickness, ash weight, breaking strength, me‐ dullary bone) were determined; (3) Results: Hens from groups with a high egg production and good eggshell quality have poorer bone quality (lower ash weight and lesser amount of medullary bone). However, Pearson’s correlation analysis shows no clearrelationship between bone and egg/eggshell parameters. (4) Conclusions: Bone and egg production/eggshell quality are independent and can be improved separately. Medullary bone has an important contribution to bone mechanical properties, being important to accumulate enough bone medullary bone early in life to maintain skeletal integ‐ rity and eggshell quality in old hens., ITC‐20161169/Ovofortis (MINECO/AEI/FEDER, UE), CGL2015‐64683‐P, RNM‐179 group (Junta de Andalucía), UCE PP 2016.05 (Universidad de Granada)

We are grateful for the support of participating volunteers, the personal of Virgen de las Nieves and San Cecilio general hospitals of Granada: Laura Campana Martin, Ma Victoria Martin Laguna and Ma Cruz Rico Prados who carried out technical work and sample preparation and Llenalia Garcia Fernandez who supervised the statistical analysis. We acknowledge the University of Granada (Spain) and the University of Guadalajara (Mexico) for the predoctoral scholarship of Laura Alejandra Vazquez Aguilar. The "NUTRAOLEUM Study" has been supported by the grant ITC-20131031 from the I + D FEDER-INTERCONNECTA (CDTI) and Junta de Andalucia, Spain". We thank ACER CAMPESTRES S.L., SAN FRANCISCO DE ASIS Coop and AGROINSUR S.L., for the funding financing provided for the development conduct of the clinical trial., Scope. The postprandial effects of virgin olive oils (VOOs) enriched with phenolic compounds and triterpenes from the olive fruit on plasma glucose and insulin (primary outcomes), and gastrointestinal hormones responses were evaluated in healthy adults. Methods and results: Single doses (30 mL) of three oils were evaluated: optimized polyphenols-rich VOO (OVOO); functional olive oil (FOO): OVOO enriched with triterpene acids; and VOO with low content of polyphenols. Postprandial plasma insulin release was lower after the intake of the FOO compared to VOO, while plasma glucose levels were lower after the intake of the VOO compared to OVOO. Matsuda's index of insulin sensitivity improved after the intake of FOO and OVOO, while the insulinogenic index and gastric inhibitory polypeptide (GIP) tended to improve after the intake of OVOO. Conclusion: The enrichment of VOOs with bioactive compounds from the olive fruit increases its benefits, improving postprandial insulin release and peripheral tissue sensitivity., University of Granada (Spain), University of Guadalajara (Mexico), I + D FEDER-INTERCONNECTA (CDTI) ITC-20131031, Junta de Andalucia European Commission, ACER CAMPESTRES S.L., SAN FRANCISCO DE ASIS Coop, AGROINSUR S.L.

All sources of funding are acknowledged in the manuscript, and authors have declared any direct financial benefits or conflict of interest that could result from publication. All appropriate ethics and other approvals were obtained for the research., We have studied in detail the changes that occur in eggshell structure and composition during a production cycle in order to better understand the deterioration of eggshell quality with hen age (at 33, 45, and 67 wk). To study changes in eggshell ultrastructure and microstructure characteristics (mammillary density, palisade layer thickness, size, and orientation of calcite crystals) and the cuticle composition, we used complementary analytical techniques such optical and electron microscopy, X-ray diffraction, and infrared spectrometry. The marked decrease in eggshell breaking strength from 5.8 Kg at 33 wk to 4.4 Kg at 67 wk (25% reduction) could not be solely explained by the modest reduction in eggshell thickness (6−10% reduction) and seems to be associate to abrupt changes in eggshell ultra- and microstructure characteristics (i.e., decreased mammillary density; increased size of crystal units), occurring in older hens. Particularly, the decrease in mammillary density reduces the attachment points of the eggshell mineral to the membranes and therefore should negatively impact eggshell mechanical properties. Also, the observed increase in the calcite crystal size making the shell could also reduce the cohesion of crystals and eggshell resistance against impacts. Additionally, there was a decrease in the amount of cuticle and internal egg quality parameters (egg albumen height) with hen age that could have a negative impact in egg safety and quality.

Results: Several single nucleotide polymorphisms on chromosome 1 between 104 and 110 Mb (galGal6) had highly significant associations with tibial breaking strength. The alternative genotypes of markers of large effect that flanked the region had tibial breaking strengths of 200.4 vs. 218.1 Newton (P < 0.002) and, in a subsequent founder generation, the higher breaking strength genotype was again associated with higher breaking strength. In a subsequent generation, cortical bone density and volume were increased in individuals with the better bone genotype but with significantly reduced medullary bone quality. The effects on cortical bone density were confirmed in a further generation and was accompanied by increased mineral maturity of the cortical bone as measured by infrared spectrometry and there was evidence of better collagen cross-linking in the cortical bone. Comparing the transcriptome of the tibia from individuals with good or poor bone quality genotypes indicated four differentially-expressed genes at the locus, one gene, cystathionine beta synthase (CBS), having a nine-fold higher expression in the genotype for low bone quality. The mechanism was cis-acting and although there was an amino-acid difference in the CBS protein between the genotypes, there was no difference in the activity of the enzyme. Plasma homocysteine concentration, the substrate of CBS, was higher in the poor bone quality genotype. Conclusions: Validated markers that predict bone strength have been defined for selective breeding and a gene was identified that may suggest alternative ways to improve bone health in addition to genetic selection. The identification of how genetic variants affect different aspects of bone turnover shows potential for translational medicine., The work was supported through an ERANET by the Biotechnology and Biological Sciences Research Council, UK grant ‘Better Bones’ BB/M028291/1, FORMAS 2014-01840, Investigación y Tecnología Agraria y Alimentaria 291815. The Roslin Institute is funded with a BBSRC Institute strategic programme grant BB/J004316/1.

A regular consumption of virgin olive oil (VOO) is associated with a reduced risk of cardiovascular disease. We aimed to assess whether the raw intake of an optimized VOO (OVOO, 490 ppm of phenolic compounds and 86 ppm of triterpenes), and a functional olive oil (FOO, 487 ppm of phenolic compounds and enriched with 389 ppm of triterpenes) supplementation (30 mL per day) during three weeks would provide additional health benefits to those produced by a standard VOO (124 ppm of phenolic compounds and 86 ppm of triterpenes) on oxidative and inflammatory biomarkers. Fifty-one healthy adults participated in a randomized, crossover, and controlled study. Urinary 8-hidroxy-2′-deoxyguanosine, plasma interleukin-8 (IL-8), and tumor necrosis factor α (TNF- α) concentrations were lower after the intervention with the FOO than after the OVOO (p = 0.033, p = 0.011 and p = 0.020, respectively). In addition, IL-8 was lower after the intervention with FOO than after VOO intervention (p = 0.002). This study provides a first level of evidence on the in vivo health benefits of olive oil triterpenes (oleanolic and maslinic acids) in healthy humans, decreasing DNA oxidation and plasma inflammatory biomarkers. The trial was registered in ClinicalTrials.gov ID: NCT02520739., The “NUTRAOLEUM Study” has been supported by the grant ITC-20131031 from the I+D FEDER-INTERCONNECTA (CDTI) and Junta de Andalucía, Spain. We thank ACER CAMPESTRES S.L., SAN FRANCISCO DE ASIS Coop. And., and AGROINSUR S.L., for the funding provided. We appreciate the support received from the workers of the clinical trials unit of the “Virgen de las Nieves” and “San Cecilio” hospitals by finding subjects for the study and for their involvement in the intervention; we thank Laura Campaña Martín, Maria Victoria Martín Laguna and Mª Cruz Rico Prados who carried out technical work, sample preparation and biomarker analyses, and Llenalia Garcia Fernández, who supervised the statistical analysis, We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)

Julio Plaza-Díaz is part of the “UGR Plan Propio de Investigación 2016” and the “Excellence actions: Unit of Excellence on Exercise and Health (UCEES), University of Granada”. We are grateful to Belen Vazquez-Gonzalez for assistance with the illustration service., The importance of gut microbiota in health and disease is being highlighted by numerous research groups worldwide. Atherosclerosis, the leading cause of heart disease and stroke, is responsible for about 50% of all cardiovascular deaths. Recently, gut dysbiosis has been identified as a remarkable factor to be considered in the pathogenesis of cardiovascular diseases (CVDs). In this review, we briefly discuss how external factors such as dietary and physical activity habits influence host-microbiota and atherogenesis, the potential mechanisms of the influence of gut microbiota in host blood pressure and the alterations in the prevalence of those bacterial genera affecting vascular tone and the development of hypertension. We will also be examining the microbiota as a therapeutic target in the prevention of CVDs and the beneficial mechanisms of probiotic administration related to cardiovascular risks. All these new insights might lead to novel analysis and CVD therapeutics based on the microbiota.

To date, pharmacokinetics of maslinic (MA) and oleanolic (OA) acids, at normal dietary intakes in humans, have not been evaluated, and data concerning their bioactive effects are scarce. We assessed MA and OA pharmacokinetics after ingestion of olive oils (OOs) with high and low triterpenic acid contents, and specifically the effect of triterpenes on endothelial function. We performed a double-blind, dose-response, randomized, cross-over nutritional intervention in healthy adults, and observed that MA and OA increased in biological fluids in a dose-dependent manner. MA bioavailability was greater than that of OA, and consumption of pentacyclic triterpenes was associated with improved endothelial function. To the best of our knowledge, this is the first time MA pharmacokinetics, and effects on endothelial function in vivo, have been reported in humans.

The present study was carried out to determine the glycemic index (GI), glycemic load (GL), insulinemic index (InI), appetite ratings and postprandial plasma concentrations of gastrointestinal hormones related to the control of food intake after the ingestion of the five most common breads consumed in Spain with different compositions and manufacturing processes. Twenty-two healthy adults participated in a randomized crossover study. The breads tested were Ordinary, Precooked-Frozen, Candeal-flour, Alfacar whites and Wholemeal. All breads portions were calculated to supply 50 g of available carbohydrates. In addition, 50 g of glucose was used as a reference. A linear mixed-effects model was used to compare data calculated for all breads with glucose load. The GI value varied from 61 for the Wholemeal, to Alfacar 68, Ordinary 76, and 78 and 86 for the Precooked-Frozen and Candeal-flour breads, respectively. Wholemeal and Alfacar had lower GI than glucose. All tested breads had a lower GL (ranged 9 to 18) compared with glucose. Wholemeal GL was similar to Alfacar, but lower than the other white breads. InI were significantly lower for all breads (ranged 68 to 73) compared with glucose, and similar among them. The intake of the Wholemeal bread led to a higher release of gastric inhibitory polypeptide compared with the Ordinary and Precooked breads and to a higher release of pancreatic polypeptide compared with the Precooked-Frozen bread. All breads affected appetite ratings similarly. In conclusion, based on GL, the Wholemeal bread would be expected to exert a favorable glycemic response.

Background Bread can contribute to the regulation of appetite. Objective The objective of this study was to investigate the appetite ratings and postprandial glucose, insulin, and gastrointestinal hormone responses related to hunger and satiety after the intake of a cereal-based bread. Methods A randomized, controlled crossover trial was conducted in 30 healthy adults (17 men and 13 women) aged 19-32 y with body mass index of 19.2-28.5. Each volunteer consumed the cereal-based bread and the control bread 2 times, with a 1-wk wash-out period, over a total of 4 sessions. The cereal-based bread contained a variety of cereal flours (wheat, oat, and spelt) and consisted of 22% dried fruits (figs, apricots, raisins, and prunes). It was also enriched with both fiber (7% from wheat cross-linked maltodextrins and pea) and protein (10-11% from wheat gluten and hydrolyzed wheat proteins). The control bread consisted of white bread with margarine and jam to control for energy density, fat, and sugar content. We measured appetite ratings using standardized visual analogue scales and glucose, insulin, and gastrointestinal hormone responses over a postprandial time of 4 h after the ingestion of each bread. Linear mixed-effects models were used to compare the areas under the curve (AUCs) for different variables. Results Consuming the cereal-based bread decreased prospective consumption more than consumption of the control bread (-5.3 ± 0.6 m · min and -4.4 ± 0.6 m · min, respectively; P = 0.02) and increased satiety more (6.2 ± 0.7 m · min and 5.2 ± 0.6 m · min, respectively; P = 0.04), although subsequent ad libitum energy intake 4 h later did not differ. Postprandial blood glucose, insulin, ghrelin, glucagon-like peptide 1 and gastric inhibitory polypeptide AUCs were lower after the ingestion of the cereal-based bread, whereas the pancreatic polypeptide AUC was higher than with the control bread (P Conclusions Consumption of the cereal-based bread contributed to appetite control by reducing hunger and enhancing satiety. In addition, consumption of this bread improved glycemic, insulinemic, and gastrointestinal hormone responses in healthy adults. This trial was registered at clinicaltrials.gov as NCT02090049.

17 Páginas; 3 Figuras; 5 Tablas, The aim of this study was to evaluate the effect of virgin olive oils (VOOs) enriched with phenolic compounds and triterpenes on metabolic syndrome and endothelial function biomarkers in healthy adults. The trial was a three-week randomized, crossover, controlled, double-blind, intervention study involving 58 subjects supplemented with a daily dose (30 mL) of three oils: (1) a VOO (124 ppm of phenolic compounds and 86 ppm of triterpenes); (2) an optimized VOO (OVOO) (490 ppm of phenolic compounds and 86 ppm of triterpenes); and (3) a functional olive oil (FOO) high in phenolic compounds (487 ppm) and enriched with triterpenes (389 ppm). Metabolic syndrome and endothelial function biomarkers were determined in vivo and ex vivo. Plasma high density lipoprotein cholesterol (HDLc) increased after the OVOO intake. Plasma endothelin-1 levels decreased after the intake of the three olive oils, and in blood cell cultures challenged. Daily intake of VOO enriched in phenolic compounds improved plasma HDLc, although no differences were found at the end of the three interventions, while VOO with at least 124 ppm of phenolic compounds, regardless of the triterpenes content improved the systemic endothelin-1 levels in vivo and ex vivo. No effect of triterpenes was observed after three weeks of interventions. Results need to be confirmed in subjects with metabolic syndrome and impaired endothelial function (Clinical Trials number NCT02520739)., The “NUTRAOLEUM Study” has been supported by the grant ITC-20131031 from the I+D FEDER-INTERCONNECTA (CDTI) and Junta de Andalucía, Spain”. We thank ACER CAMPESTRES S.L., SAN FRANCISCO DE ASIS Coop and AGROINSUR S.L., for the funding provided. The authors thank Pilar Jiménez, Alberto Guarnido, María Molina and Elizabeth García for administering the questionnaires to the subjects; Isabel Mérida, Isabel Hinojosa, Agustín Martín García, and María Luz Abarca for collecting the biological samples; and Victoria Martín Laguna and Laura Campaña Martín for aliquoting the samples. The authors also thank María Cruz Rico Prados for sample analysis and Llenalia M. García Fernández for her contributions to the statistical analysis.

Overweight and obesity are considered a world pandemic of the 21st century. Obesity is a complex multifactorial disease influenced by lifestyle, behavioral, environmental, and genetic factors. Metabolic syndrome (MetS) is a cluster of associated metabolic disturbances which tend to coexist, commonly represented by the combination of obesity (especially abdominal obesity), hyperglycemia, dyslipidemia, and hypertension. MetS is associated with an increased risk of cardiovascular diseases (CVDs), characterized by abnormal cytokine production that leads to endothelial dysfunction. Oxidative stress is associated with CVDs and their related risk factors, such as obesity and type 2 diabetes, and with other chronic degenerative processes such as cancer and other aging-related diseases. Epidemiological evidence indicates that polyphenol-rich diets, such as the Mediterranean diet, are associated with antioxidant, antiinflammatory, and antiobesity effects and with the reduction of risk factors for MetS and CVDs. Polyphenols can be found in nature, mainly in plants and vegetables. Probably the best known beneficial effect of polyphenols is their antioxidant capacity, which is capable of reducing reactive oxygen species and various organic substrates and minerals. In this chapter we explain the nature of polyphenols and their classification, and elucidate their potential mechanism of action and the current evidence in regard to obesity and MetS.

Food safety of table eggs is vital since many pathogens can contaminate the unfertilized egg, leading to increased risk of foodborne illness for consumers. The eggshell cuticle is the first line of defense to restrict the entry of egg-associated pathogens, such as Salmonella Enteritidis. The thickness and completeness of coverage of the cuticle layer are heritable traits that are strongly associated with egg resistance to bacterial penetration. The present study characterizes the chemical composition of the eggshell cuticle and structure of pore plugs from table eggs. Eggs collected from both brown and white egg laying Lohmann flocks (early, mid, and late lay) were either unwashed or washed. Pore plugs were characterized using scanning electron microscopy (SEM), and elemental composition was determined using energy-dispersive x-ray spectroscopy (EDS). SEM observations confirmed that the plug formed by the cuticle layer within the eggshell pore remains firmly lodged throughout the commercial washing process. The eggshell thickness and cuticle pore length visualized in brown eggs was significantly higher than in white eggs in hens of all ages. EDS analysis revealed that the pore inner surface was enriched in phosphorus and chemically different from the surrounding bulk eggshell mineral. Detailed assessment of the cuticle chemical composition was performed by Fourier transform infrared spectroscopy (FTIR). Washing of eggs removed cuticle from the eggshell surface. There was a trend of lower cuticle coverage with increasing hen age for white eggs. A significant reduction in the amount of proteins and phosphates and polysaccharides was observed in the cuticle of brown unwashed eggs with hen age. In white unwashed eggs, amides and lipids decreased with hen age; by contrast, the amount of sulfate was highest at mid-lay. The results from our research will assist selective breeding programs that target cuticle integrity and pore plug stability to enhance egg resistance to pathogen penetration and improve food safety.

Olive oil is rich in several minor components like maslinic (MA) and oleanolic (OA) acids which have cardioprotective, antitumor, and anti-inflammatory properties. In order to assess the health benefits in humans provided by the olive oil triterpenes (MA and OA), suitable analytical methods able to quantify the low concentrations expected in human fluids are required. In this study, the LC-MS/MS quantification of both OA and MA in plasma and urine has been evaluated. The plasmatic method is based on the direct determination of the analytes. The urinary detection requires more sensitivity which was reached by derivatization with 2-picolylamine. Additionally, the urinary species present after MA and OA ingestion were evaluated by the direct detection of several phase II metabolites previously synthesized. Our results showed that OA is metabolized as both sulfate and glucuronide conjugates whereas MA is mainly excreted as glucuronide. Based on this information, the method for the urinary detection of MA and OA involved an enzymatic hydrolysis. Both plasmatic and urinary methods were validated with suitable precision and accuracy at all tested levels. Required sensitivity was achieved in both matrices. Up to our knowledge, this is the first method able to quantify the low concentration levels of triterpenes present in urine. Samples from two healthy volunteers who received virgin olive oils with different triterpenes content were analyzed. Some preliminary clues on the metabolic disposition of OA and MA after olive oil intake are provided.

9 Páginas; 3 Tablas; 1 Figura, Background Virgin olive oil, a recognized healthy food, cannot be consumed in great quantities. We aim to assess in humans whether an optimized virgin olive oil with high phenolic content (OVOO, 429 mg/Kg) and a functional one (FOO), both rich in phenolic compounds (429 mg/Kg) and triterpenic acids (389 mg/kg), could provide health benefits additional to those supplied a by a standard virgin olive oil (VOO). Methods/design A randomized, double-blind, crossover, controlled study will be conducted. Healthy volunteers (aged 20 to 50) will be randomized into one of three groups of daily raw olive oil consumption: VOO, OVOO, and FOO (30 mL/d). Olive oils will be administered over 3-week periods preceded by 2-week washout ones. The main outcomes will be markers of lipid and DNA oxidation, inflammation, and vascular damage. A bioavailability and dose-response study will be nested within this sustained- consumption one. It will be made up of 18 volunteers and be performed at two stages after a single dose of each olive oil. Endothelial function and nitric oxide will be assessed at baseline and at 4 h and 6 h after olive oil single dose ingestion. Discussion For the first time the NUTRAOLEUM Study will provide first level evidence on the health benefits in vivo in humans of olive oil triterpenes (oleanolic and maslinic acid) in addition to their bioavailability and disposition., The NUTRAOLEOUM Study has been supported by the grant ITC-20131031 from the FEDER-INTERCONNECTA (CDTI) and Junta de Andalucía, Spain. We thank ACER CAMPESTRES, San Francisco de Asis Coop. And., and AGROINSUR SL, for providing the materials and processes for obtaining the olive oils used in the Project. These sources do not have any rol or authority in the in study design; collection, management, analyses, and interpretation of data; writing or submission of the report.