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BACKGROUND: Animal handling practices are one of the factors majorly affecting animal metabolism prior to slaughter. This phenomenon increases the occurrence of meat quality defects such as dark cutting-beef, causing high economical losses in the meat industry. Under this framework, the assessment of apoptosis onset in post mortem muscle was proposed as a novel approach to reveal biochemical characteristics in several Spanish bovine breeds (Asturiana de los Valles, Retinta and Rubia Gallega) managed under different production systems (intensive versus semi-extensive) and transport/lairage conditions (mixing versus not mixing with unfamiliar animals). To do so, the activities of initiator caspase 9 and executioner caspases 3/7 were determined in Longissimus thoracis et lumborum muscle at three early post mortem times (2, 8, and 24 h). RESULTS: Breed effect and transport/lairage conditions were the most relevant factors that influenced both caspase activities over post mortem time, showing Rubia Gallega breed a completely different behavior compared to Asturiana de los Valles and Retinta breeds. Moreover, it is postulated that apoptosis cascade is initiated via the activation of caspase 9 under hypoxic or metabolic stress followed by the activation of executioner caspases 3/7. CONCLUSIONS: Assessment of apoptosis on post mortem muscle can be a novel approach to study the influence of animal handling on muscle metabolism and post mortem cell death and its consequences on meat quality traits., Claudia Fuente-Garcia thanks the Department of Economic Development & Infrastructures of the Basque Government for the doctoral fellowship. This work was funded by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (RTA2014-00034-C04), Agencia Estatal de Investigación (RTI2018-096162-R-C22) and FEDER funds.

Seventy-eight calves from Asturiana de los Valles, Retinta, and Rubia Gallega breeds, under extensive and intensive farm systems and animal mixing and non-mixing conditions, and during the transport and lairage in slaughterhouses, were studied. This research aimed to study the effect of breed, farm system and mixing conditions on serum biomarkers (cortisol, lactate, glucose, serum amyloid A, haptoglobin, and C-reactive protein) and their relationship with pHu at slaughter time, and to evaluate the response of the serum biomarkers of calves throughout fattening period. Moreover, this study aims to evaluate the response of the biomarkers in each breed during the fattening period. At slaughter time, cortisol and lactate were affected by BreedxFarm; Retinta showed the opposite pattern to the others and revealed the highest glucose in extensive farm systems. Rubia Gallega in mixing revealed the highest Amyloid A and haptoglobin. Extensive calves in mixing conditions showed the highest glucose. There was a relationship among the variables cortisol, lactate, Amyloid A, and pHu. Slaughter time was a major stressor, and the stress response was mainly affected by breed. At slaughter, several biomarkers should be considered., This research was funded by INIA-MINECO (Spain), under RTA2014-00034-C04-00 project, and FEDER funds.

A full-randomized block design was used for the study of the FA composition and meat quality parameters, considering ageing time as a split-plot factor. Chemical and fatty acid composition of steaks (longissimus thoracis and lumborum muscle) from 15 month old semiextensively reared Hispano-Bretón horses were characterized (day 0), and the effect of vacuum ageing (0, 7, 14 and 21 days) on several meat quality parameters (pH, instrumental color and texture and cook loss) was determined. The average fat content of horse loin was 3.31%, and the n-3 polyunsaturated fatty acid content, although higher than in ruminant meats, suggested that the finishing on a high-grain diet limited muscle n-3 accumulation. Results revealed that ageing affected all meat quality measurements; color started to turn brownish at 14 days of ageing, with a decrease in redness but not in yellowness. Tenderness improved during the first two weeks, and the Warner-Bratzler shear force scores showed that meat aged for 7 days could be considered as ‘intermediate tender’. Under the present study conditions, an ageing period between 7 and 14 days is recommended for an optimum horse meat quality., This research was funded by the Basque Government (grant IT944-16) that also funded the postdoctoral grant of L.M. Department of Economic Development and Infrastructures of the Basque Government is acknowledged for the fellowship of L.R.B.

International audience; Ahmed Ouali was born on October 4, 1948 in Tigzirt, Tizi-Ouzou, Algeria. In 1952, he moved with his parents to Montluçon, France. In 1974, he was trained and graduated with a bachelor's degree in Biochemistry at the University of Lyon. He then, in 1976, earned a joint Ph.D. in Animal Science at the University of Blaise Pascal (Clermont-Ferrand) where he studied at the National Institute of Agricultural Research (INRA, Theix). The title of his doctorate thesis was “The role of muscle proteases on meat tenderization”. Subsequently, he was employed in a private laboratory for medical analysis from 1976 to 1978 and thereafter at the Meat Research Laboratory group at INRA, Theix as a permanent researcher. In 1990, he was appointed as a research director and led the “Biochemistry and Functions of Muscle Proteins” unit for 8 years. The Meat Research Station focused their research on many topics including colour and protein oxidation; enzymology and tenderness; and muscle protein functionalities. During his entire scientific career at INRA, but before his retirement on October 2013, Ahmed was living in Clermont-Ferrand, the city of the famous volcanic chain of the Puy-de-Dôme, with his wife Anne-Marie with whom he had two lovely children: Armelle (41 years) and Gäel (38 years). In 2019, they moved to their new house in Montpellier in the South of France.

Estimates of quantitative proteomic distance between populations have not been reported to date. Here, quantitative proteomic distances between three Spanish bovine breeds (Asturiana de los Valles, AV; Retinta, RE; and Rubia Gallega, RG) were estimated from two-dimensional electrophoresis profiles of meat samples of longissimus thoracis muscle at 2 h post-mortem. Statistically significant distances were detected between AV/RG and the most genetically different RE breed, using the novel QD measure of quantitative proteomic distance. In total, 18 differentially abundant myofibrillar and sarcoplasmic proteins/isoforms contributing to proteomic distances between breeds were confidently identified by tandem mass spectrometry. The fast skeletal myosin regulatory light chain 2 followed by other five interacting proteins exhibited the most pronounced relative change between breeds. In addition, most differentially represented proteins could be associated with variations in meat tenderness. Therefore, they could be candidate biomarkers for molecular breeding programs and authentication of the three Spanish beef breeds., This work was supported by the Instituto Nacional de Investigación y Tecnología Agraria (INIA, RTA 2014-00034-C04), Spain; and by a predoctoral fellowship of the Xunta de Galicia (Spain) and the European Union (ESF) to R. Rodríguez-Vázquez.

Different bovine breeds and production systems are used worldwide, giving rise to differences in intrinsic and extrinsic characteristics of beef. In order to meet the consumer requirements, new approaches are currently being developed to guarantee tenderness, taste, and juiciness of beef. However, the final consumer perception is complex, and it is also affected by several interrelated variables. This study aimed to evaluate the physicochemical parameters and sensory profile of three Spanish cattle breeds under different livestock production systems (extensive and intensive) and pre-slaughter handling conditions (mixing and not mixing with unfamiliar individuals at pre-mortem time). Meat samples from each group were also studied at different ageing times (7 and 14 days). Regarding sensory attributes, twelve panelists assessed meat samples and an exhaustive statistical analysis was carried out. The most evident and strongest effect was the breed type, allowing a great differentiation among them using principal components and discriminant analysis. The livestock production system was the second most important parameter, significantly affecting odor, flavor, and textural profile (fibrousness). It can be concluded that there were marked differences in the traits of these beef that could be modified by other factors in order to fulfill consumer tastes., Authors are grateful to RTA 2014-00034-C04-00 (INIA-MINECO) and FEDER funds for the financial support. Special thanks for financial support from the Xunta de Galicia and the European Union (ESF) for supporting Raquel Rodríguez-Vázquez pre-doctoral scholarship. Daniel Franco and José M. Lorenzo are members of the HealthyMeat network, funded by CYTED (Ref. 119RT0568).

The aim of the present study was to evaluate the differences in salivary proteome at rest and in response to an acute exercise in men and women. For this, unstimulated whole saliva samples in rest and after a bout resistance exercise leading to failure (ELF) of both men and women (n = 5 for each sex) were subjected to isobaric Tandem Mass Tags (TMT) labelling followed by LC-MS/MS. A total of 274 proteins were identified and met the inclusion criteria. 16 proteins were modulated for the interaction sex*exercise, 6 were modulated because of the exercise, and 65 were differentially expressed between men and women at rest. In conclusion, these results indicate sex-related differences in the salivary proteome at rest and after an acute exercise, pointing out possible candidate biomarkers for sports performance and allowing further knowledge of the physiological processes occurring during ELF., LFM was granted with a predoctoral contract ‘FPU’ from the University of Murcia, Spain. AT has a post-doctoral fellowship “Ramón y Cajal” supported by the “Ministerio de Economía y Competitividad”, Spain. This work was supported by a grant from the Program for Research Groups of Excellence of the Seneca Foundation, Murcia, Spain (grant 19894/GERM/15) and European Commission ERA chair FP7 grant (VetMedZg #621394).

Although Peptidomics is a discipline complementary to proteomics, since nowadays both mainly rely on analytical strategies based on mass spectrometry, there are fundamental differences. In this chapter, we discuss these differences along with the application of these technologies for the study of the different stages of meat production, from storage to processing to unravel mechanisms that will allow reaching high quality and safer meat products. The use of Peptidomics and the related high throughput technologies, now relying on mass spectrometry but once also on N-terminal sequencing, are discussed. Clear examples are provided dealing with relevant studies on meat proteolysis and peptide generation occurring during ageing, as well as those produced during ripening of meat products by endogenous and microbial enzymes. Also the involvement of this phenomenon in the development of taste active compounds is addressed. Finally, the application of novel Omics technologies on bacterial identification in food for diagnosis and safety purposes is presented, putting emphasis on their potential advantages and future perspectives. Fil: Fadda, Silvina G.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Sentandreu, Enrique. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; España Fil: Sentandreu Vicente, Miguel Angel. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; España

Epub 2018 May 9; International audience; Protein biomarkers of meat tenderness are known to be of primary importance for the prediction of meat quality, and hence, industry profitability. Proteome analysis was performed on meat from 8 Main Anjou beef cattle, previously classified as tender or tough meats by Warner Bratzler shear force measurements. Myofibrillar fraction of Longissimus thoracis muscle was separated by a novel fractionation approach based on liquid isoelectric focusing (OFFGEL) and further analyzed by SDS-PAGE and liquid chromatography coupled to tandem mass spectrometry. Obtained OFFGEL fraction profiles were reproducible allowing the comparison of both meat qualities and revealing 7 protein bands capable to discriminate between tender and tough samples. The proteins present in these bands were troponin T, Heat Shock protein beta-1, creatine kinase, actin, troponin C, myosins 1 and 2 and myozenin-1. The latter protein has not been previously reported as a marker of meat tenderness. This study introduces an innovative proteomic approach for the study of muscle proteome. The fact of obtaining fractions in liquid state after OFFGEL fractionation allows for a faster analysis of proteins by mass spectrometry, being an interesting alternative to more classical proteomic approaches based on two dimensional gel electrophoresis (2-DE).

The hydrolysis of myofibrillar proteins during fermentation of sausage models by an autochthonous starter culture was investigated. In order to provide a whole map of the generated products, proteomic and peptidomic were used andcomplemented with the amino acid profile. Beaker sausages (BS) were used as models which were inoculated or not with Lactobacillus curvatus CRL705 and Staphylococcus vitulinus GV318 as starter cultures. The hydrolysis of actin, myosin light chain 1/3 (MLC 1/3), myosin regulatory light chain-2 (MRLC-2) and myosin heavy chain (MHC) were evidenced by two-dimensional gel electrophoresis (2-DE). In addition, a total of 33 peptides arisen from troponin T, MRLC-2 and particularly from actin were identified by LC-MS/MS. These results showed that the starter culture significantly enhanced the proteolysis of the proteins named above, even when the endogenous enzymes induced a clear breakdown. L. curvatus CRL705 highly enriched both peptide pattern and amino acid concentrations. When the autochthonous starter culture was inoculated, although proteolysis was remarkably reinforced, a reduction in peptide and amino acid composition was observed. Regarding actin primary structure, three regions of this proteinwere highly susceptible to degradation by the starter culture. Additionally, the essential role of exopeptidases- from meat and bacteria- in diversity of actin peptides during fermentation was shown. This study improved the knowledge of the proteolysis of myofibrillar proteins and the involved enzymes, as well as, completed the previously reported degradation of sarcoplasmic proteins by the same autochthonous starter culture. The singular peptides and amino acids pattern generated might contribute to the uniqueness of produced fermented sausages while they may be used as quality markers. Fil: López, Constanza María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Sentandreu Vicente, Miguel Angel. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; España Fil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Fadda, Silvina G.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina

Postmortem (PM) meat tenderization is a complex biochemical process that involves multiple endogenous proteolytic enzymes and that is not completely understood. Cell death by apoptosis is recently proposed as a novel mechanism in this process. The signification of the caspases implication in muscle proteins degradation is that the cells are dying through apoptosis, a major cell death programme. Our purpose was therefore to verify this statement using different well known hallmarks of the apoptotic process including cell shrinkage, phosphatidylserine (PS) externalization. Cells start to shrink few minutes after animal death and reach a maximum shrinkage about 24 h postmortem while they showed the typical rounding appearance of apoptotic cells. Externalization of PS is detectable within 1 h postmortem and increased gradually with time. The present work is therefore the first to provide direct evidence supporting the onset of apoptosis in post-mortem muscle. The present findings should lead us to reconsider as a whole, the mechanisms contributing of the development of meat qualities (tenderness, flavor, juiciness …) by a detailed analysis of the apoptosis associated biochemical and physiological modifications taking place within the cells.

International audience; In living cells, after activation, protein inhibitors constitute the last step of proteases activity regulation. This review intends to provide original information about a group of bovine muscle serine proteases inhibitors belonging to the Serpin superfamily and characterized at the gene and protein level. This report is the only one and the first to provide much information on this group of proteases inhibitors of the serpin type and their potential biological functions. Amongst the eight genes identified in bovine, three serpins were purified from the muscle tissue and characterized. These are two members of the bovSERPINA3 family, i.e., bovSERPINA3-1 and A3-3, and the last one is antithrombin III (AT-III or BovSERPINC1). BovSERPINA3 family comprises at least eight protein members encoded by different genes mapped on chromosome 7q23–q26 cluster. BovSERPINA3-1 and A3-3 were shown to locate within muscle cells and are cross-class inhibitors strongly active against trypsin as well as against human initiator and effector caspases 8 and 3. They constitute a key apoptosis control in mammals. They were thus expressed in proliferating and confluent myoblasts phases where cells must be alive but not in myotubes. Antithrombin III inhibits trypsin and, in a heparin dependent manner, thrombin. AT-III and its mRNA were expressed in muscle cells and in differentiating primary myoblasts in culture.

The present report overviews a new family of bovine serpins able to inhibit pseudo-irreversibly initiator and effector caspases, a group of cysteine proteases in charge of cell dismantling during apoptosis, a finely regulated cell death process. The 8 members identified at the gene level showed a high homology with human SERPINA3 and were therefore designed bovSERPINA3-1 to A3-8. At least six of them are able to inhibit caspases. Two of them (bovSERPINA3-1 and A3-3) have been purified from bovine muscle and extensively investigated during these last years. After a general presentation of the serpin superfamily, the kinetic aspects of their interaction with human cas-pases 3 and 8 were studied and findings obtained suggest that caspases could be their target enzymes in living cells. In muscle and primary myoblast in culture, they showed an intracellular localization and because of their high level in blood, they can be exported. Two biological functions (potential regulator of apoptosis and expression during myoblast differentiation) were investigated and it was concluded that they are very likely a efficient regulator of apoptosis, a proposal supported by their high expression in proliferating myoblast (cell survival is essential during this differentiation phase) but not in myotubes.

Serpins are a superfamily of structurally conserved proteins. Inhibitory serpins use a suicide substrate-like mechanism. Some are able to inhibit cysteine proteases in cross-class inhibition. Here, we demonstrate for the first time the strong inhibition of initiator and effector caspases 3 and 8 by two purified bovine SERPINA3s. SERPINA 3-1 (uniprotkb: Q9TTE1 ) binds tighly to human CASP3 (uniprotkb: P42574 ) and CASP8 (uniprotkb: Q14790 ) with kass of 4.2 × 105 and 1.4 × 106 M−1 s−1, respectively. A wholly similar inhibition of human CASP3 and CASP8 by SERPINA3-3 (uniprotkb: Q3ZEJ6 ) was also observed with kass of 1.5 × 105 and 2.7 × 106 M−1 s−1, respectively and form SDS-stable complexes with both caspases. By site-directed mutagenesis of bovSERPINA3-3, we identified Asp371 as the potential P1 residue for caspases. The ability of other members of this family to inhibit trypsin and caspases was analysed and discussed. Structured summary MINT- 7234656 : CASP8 (uniprotkb: Q14790 ) and SERPINA3-1 (uniprotkb: Q9TTE1 ) bind (MI: 0407 ) by biochemical (MI: 0401 ) MINT- 7234634 : SERPINA3-3 (uniprotkb: Q3ZEJ6 ) and CASP3 (uniprotkb: P42574 ) bind (MI: 0407 ) by biochemical (MI: 0401 ) MINT- 7234663 : CASP8 (uniprotkb: Q14790 ) and SERPINA3-3 (uniprotkb: Q3ZEJ6 ) bind (MI: 0407 ) by biochemical (MI: 0401 ) MINT- 7234625 : SERPINA3-1 (uniprotkb: Q9TTE1 ) and CASP3 (uniprotkb: P42574 ) bind (MI: 0407 ) by biochemical (MI: 0401 )

In the present work, a new endopin-like serpin designed mEndopin 1B was purified from bovine muscle. Biochemical characterizations (amino acid sequencing and Maldi-Tof mass spectrometry peptide mapping) demonstrated that the purified protein is different from the previously described Endopin 1, renamed mEndopin 1A. The genes and cDNA of both endopins were characterized. The cDNA sequence of mEndopin 1B encodes a predicted protein of 411 amino-acids with a molecular mass of 43808Da. The mEndopin 1B gene comprised four coding exons and an additional 5' untranslated exon. The reactive site sequence of mEndopin 1B is somewhat different from that of mEndopin 1A. Nevertheless, both serpins have a similar peptidase inhibitory pattern against examined proteases (elastase, trypsin, plasmin and chymotrypsin). The high expression of both mEndopin 1A and 1B in bovine serum and tissues and their high efficiency to inhibit elastase (k(ass) approximately 10(6)-10(7) M(-1) s(-1)) suggested that these serpins might play a major role in inflammatory processes.

In the present work, an endopin-like elastase inhibitor was purified for the first time from bovine muscle. A three-step chromatography procedure was developed including successively SP-Sepharose, Q-Sepharose and EMD-DEAE 650. This procedure provides about 300 mu g of highly pure inhibitor from 500 g of bovine diaphragm muscle. The N-terminal sequence of the muscle elastase inhibitor, together with the sequence of a trypsin-generated peptide, showed 100% similarity with the cDNA deduced sequence of chromaffin cell endopin 1. Hence, the muscle inhibitor was designated muscle endopin 1 (mEndopin 1). mEndopin 1 had a molecular mass of 70 kDa, as assessed by both gel filtration and SDS/PAGE. According to the association rates determined, mEndopin 1 is a potent inhibitor of elastase (k(ass) = 2.41 x 10(7) M-1 . s(-1)) and trypsin (k(ass) = 3.92 x 10(6) M-1 . s(-1)), whereas plasmin (k(ass) 1.78 x 10(3) M-1 . s(-1)) and chymotrypsin (k(ass) = 1.0 x 10(2) M-1 . s(-1)) were only moderately inhibited. By contrast, no inhibition was detected against several other selected serine proteinases, as well as against cysteine proteinases of the papain family. The cellular location of mEndopin in muscle tissue and its tissue distribution were investigated using a highly specific rabbit antiserum. The results obtained demonstrate an intracellular location and a wide distribution in bovine tissues.

International audience; For consumers, tenderness is the most important sensory attribute of beef meat and, though to a lesser extent, of pork. Tenderness is therefore by far the most common cause of its unacceptability. The major challenge for the beef industry is to evaluate the toughness of the meat as soon as possible after death. In this context, the aim of the present work was to develop an equation to predict the myofibrillar ultimate resistance of raw meat. The study was done on the Longissimus muscle from twenty three 19 months-old Charolais bulls grown in the same INRA farm. Muscles excised within 1 h post-mortem were vacuum packed and stored at 15 degrees C during 24 h and then transferred to 4 degrees C until used. The activities of lactate dehydrogenase, citrate synthase and myofibrillar Mg-Ca dependent ATPase, the levels of lactate dehydrogenase enzyme, myoglobin, myosin types 1, IIa and IIb, cysteine and serine peptidase inhibitors, the pH, the osmolarity, the expressible juice, mu-calpain, in-calpain and calpastatin and meat toughness were measured. According to the physical method used here, the force measured on raw meat represents the resistance of the myofibillar structure. Stepwise linear regression was used to determine the best equation (p < 0.05) for predicting toughness at 6 days post-mortem. A 6-variables predictive equation including serine peptidase inhibitors (partial R-2 = 0.4), the rate (partial R-2 = 0.25), and the extent of pH decline (partial R-2 = 0.03), the at death LDH activity (partial R-2 = 0.24), the extent of increase in osmotic pressure (partial R-2 = 0.13), and the rate of mu-calpain activity loss (partial R-2 = 0.09), explained 70% of the variability in meat toughness at 6 days post-mortem. This equation was developed from 20 animals and the other 3 animals, chosen randomly, were used to validate it. The absolute need for a predictive model of meat toughness and the nature of the serine peptidase inhibitors together with their potential target enzymes are discussed.

Angiotensin I-converting enzyme (ACE) inhibitory activity of peptides derived from the hydrolysis of sarcoplasmic and myofibrillar porcine proteins by the action of Lactobacillus sakei CRL1862 and Lactobacillus curvatus CRL705 (whole cells + cell free extracts) was investigated at 30 °C for 36 h. The protein hydrolysates were subjected to RP-HPLC in order to fractionate the extracts for further evaluate the ACE inhibitory activity of the collected peptide fractions. Bioactive fractions were only found from the hydrolysis of sarcoplasmic proteins by both assayed lactic acid bacteria. Identification of peptides contained in these bioactive fractions was carried out by tandem mass spectrometry using a nanoLC-ESI-QTOF instrument and the mascot seach engine. A total of eighteen peptides were characterized from the two most active fractions obtained from the hydrolysis made by L. sakei CRL1862. Regarding L. curvatus CRL705, its proteolytic action was able to generate thirty and twenty peptides, which were identified in the two most active fractions, respectively. It is worth noting that the sequence FISNHAY was generated by the proteolytic action of the two species. Sequence similarity analyses between the peptides identified in this study and those previously identified as ACE inhibitory peptides and detailed in the BIOPEP database were outlined. Results suggest that lactic acid bacteria selected in this study were able to generate peptides with ACE inhibitory activity, thus having potential to be used in the development of functional fermented products. Fil: Castellano, Patricia Haydee. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Fil: Aristoy, María Concepción. Consejo Superior de Investigaciones Cientificas. Instituto de Ciencia y Teconologia de Alimentos y Nutricion; España; Fil: Sentandreu, Miguel Ángel. Consejo Superior de Investigaciones Cientificas. Instituto de Ciencia y Teconologia de Alimentos y Nutricion; España; Fil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Fil: Toldrá, Fidel. Consejo Superior de Investigaciones Cientificas. Instituto de Ciencia y Teconologia de Alimentos y Nutricion; España