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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.

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.

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)

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.

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.