Déterminisme biologique de la variabilité de la fonte lipidique à la cuisson du foie gras de canard

Les objectifs de ce travail sont d'identifier les mécanismes biologiques impliqués dans le déterminisme de la variabilité de la fonte lipidique à la cuisson du foie gras de canard, et de définir des marqueurs de la fonte. Pour répondre à ces objectifs, nous avons choisi de conduire une caractérisation du tissu hépatique, selon son comportement ultérieur à la cuisson, basée sur une approche intégrée combinant de la biochimie, de la protéomique et de l'histologie. D'une part, l'expression différentielle des protéines hépatiques à l'abattage des animaux et au cours de la réfrigération, semble indiquer que le niveau de fonte pourrait être relié à des stades de développement de stéatose différents. Les animaux du groupe ‘Fonte faible' se distinguent de ceux du groupe ‘Fonte élevée' par un ‘profil d'accrétion' avec la surexpression de protéines du métabolisme énergétique par exemple, indiquant des mécanismes de synthèse et d'accumulation de lipides. A l'inverse, les animaux du groupe ‘Fonte élevée' présentent la surexpression de protéines de réponse au stress, ce qui pourrait indiquer un stade de stéatose plus avancé. Au cours de la réfrigération, la diminution globale de l'expression des protéines semble plus marquée dans le groupe ‘Fonte faible'. Dans le groupe ‘Fonte élevée', la surexpression de protéines du cytosquelette après réfrigération suggère un niveau de protéolyse plus important. D'autre part, les observations histologiques montrent des modifications de la morphologie des gouttelettes lipidiques dans le foie gras cru et au cours de la cuisson. Il semble que la fonte soit d'autant plus importante quand des fusions de gouttelettes lipidiques surviennent dans le produit cru. De plus, lors de la cuisson, les observations sur foies gras du groupe ‘Fonte élevée' révèlent une densification de la matrice non lipidique plus importante. Ces changements morphologiques et structuraux pourraient correspondre à un tissu plus fragile et donc plus sensible à la fonte lipidique. L'ensemble des résultats acquis dans ce projet permet de formuler une hypothèse sur le déterminisme biologique de la fonte lipidique même si certains mécanismes identifiés restent à confirmer. De plus, la relation entre l'aptitude à la stéatose et la qualité technologique du foie gras est une piste à explorer afin de mieux comprendre la variabilité de la fonte lipidique à la cuisson observée lors de la cuisson du foie gras de canard. ABSTRACT : This work aims to identify the biological mechanisms involved in the determinism of variability of fat loss during cooking of duck ‘foie gras'. To achieve this objective, we carried out a characterization of liver tissue, according to its later behaviour during cooking, and based on an integrated approach combining biochemistry, proteomic and histology. On the one hand, the differential expressions of proteins early post mortem and during chilling indicate a different state of steatosis between the low and high fat loss groups. Livers with low fat loss during cooking were still in anabolic processes with regards to energy metabolism and protein synthesis, whereas livers with high fat loss during cooking developed cell protection mechanisms. The overall expression of proteins in the low ionic strength fraction was lower after chilling which revealed a down regulating effect of chilling on biological processes. In the high ionic strength fraction, the results showed an over expression after chilling of proteins from cytoskeleton and their associated proteins. This suggests that the variability of technological yield observed in processing plants could be explained by different state of ageing of fatty livers during chilling, most likely associated to different proteolytic pattern. On the other hand, the histological observations showed that, in raw livers, the lipid droplets were nearly spherical while after cooking, they were larger and lost their spherical shape. We also observed a decrease in the number of droplets after cooking, probably due to droplet fusion caused by the heat treatment. Fat loss during cooking was higher when there was more fusion of lipid droplets before cooking. Furthermore, it seemed that the effect of cooking on liver tissue was different between the two fat loss groups. These morphological modifications could mark a fragility of the tissue associated with a higher fat loss during cooking. All the results acquired in this work allowed us to draw a hypothesis on the biological determinism of cooking losses eventhough some of the identified mechanisms still need to be confirmed. The relationship between the potential to develop steatosis and the technological quality of ‘foie gras' deserves further studies in order to better understand the variability in fat loss during cooking observed in processing plants

Approche métabolomique pour la recherche de marqueurs métaboliques de la qualité technologique des foies gras

La maîtrise de la fonte lipidique des foies gras à la cuisson représente aujourd’hui un enjeu majeur pour les acteurs de la filière. L’amélioration des différentes étapes : élevage-gavage, abattage et transformation des foies a permis de réduire les pertes lipidiques à la cuisson. Cependant une part de la variabilité de ce paramètre reste mal contrôlée. Pour mieux comprendre cette variabilité, les caractéristiques intrinsèques de deux groupes de foies gras avec des valeurs extrêmes de rendement technologique ont été comparées. Leurs métabolites hydrophiles ont été analysés en RMN du proton, puis comparés. Ces résultats ont renforcé l’idée que i) le catabolisme protéique est plus élevé dans les foies avec des rendements technologiques faibles que dans ceux avec des rendements élevés et ii) que le stress oxydatif semble moindre dans les foies avec des rendements technologiques élevés. Les membranes plasmiques pourraient avoir une meilleure intégrité ce qui expliquerait la plus grande rétention de lipides lors de la cuisson dans ce groupe de foie gras

La fonte lipidique du foie gras à la cuisson

La connaissance des facteurs biologiques, zootechniques et technologiques ainsi que des mécanismes qui contrôlent le taux de fonte des foies gras à la cuisson est primordiale pour la maîtrise du rendement technologique des foies gras. En effet, le taux de fonte à la cuisson a des répercussions sur les qualités organoleptiques des foies gras et sur les performances des unités de production

Metabolomic study of fatty livers in ducks: Identification by 1H-NMR of metabolic markers associated with technological quality

International audienceThe control of fatty liver fat loss during cooking is a major issue. Previous studies showed that fat loss was influenced by bird production factors and liver technological treatments. However, part of the variability in fat loss remained uncontrolled. To provide enhanced insights into the determinism of fatty liver quality, liver hydrophilic metabolite profiles were measured by nuclear magnetic resonance of the proton (1H-NMR). The study aimed at i) comparing fatty livers with extreme fat loss values and ii) at characterizing the effect of postmortem evolution of livers during chilling. A group of 240 male mule ducks (Cairina moschata × Anas platyrhynchos) was reared and overfed. Their livers were sampled at 20 min and 6 h postmortem. Of these birds, 2 groups of ducks were built with extreme values on the technological yield (TY; TY = 100 − % fat loss; the low-fat-loss group, TY = 89.9%, n = 13; and the high-fat-loss group, TY = 68.3%, n = 12, P < 0.001). The 1H-NMR analyses showed that the high-fat-loss livers were more advanced in postmortem biochemical and structural changes than low-fat-loss livers early postmortem. The high-fat-loss livers were characterized by hydrolysis of glycogen into glucose, worse integrity of cell membrane with diminution of compounds of phospholipids, and higher catabolic processes. The accelerated postmortem processes may be the origin of the differences in fat loss during cooking. During the early postmortem period, the adenosine triphosphate amount in liver cells was strongly reduced and lipolysis of triglycerides seemed to be enhanced. The glycogen stored in liver was first converted into glucose, but contrary to what happens in postmortem muscles, glucose was not converted into lactate

Understanding the Effects of a Tannin Extract on Forage Protein Understanding the Effects of a Tannin Extract on Forage Protein

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Chemical reactivity of nitrite and ascorbate in a cured and cooked meat model implication in nitrosation, nitrosylation and oxidation

International audienceNitrite, added to cured meat for its bacteriological and technological properties, is implicated in the formation ofnitroso compounds (NOCs), such as nitrosylheme, nitrosamines and nitrosothiols, suspected to have a potentialimpact on human health. The mechanisms involved in NOC formation are studied in regard with the dose–response relationship of added nitrite and its interaction with ascorbate on NOC formation in a cured andcooked meat model. The impact of a second cooking stage on nitrosation was evaluated. The addition of nitrite inthe cured and cooked model promoted heme iron nitrosylation and S-nitrosation but not N-nitrosation. Nitritereduced lipid oxidation without an additional ascorbate effect. The second cooking sharply increased thenitrosamine content while the presence of ascorbate considerably lowered their levels and protected nitrosothiolsfrom degradation. This study gives new insights on the chemical reactivity of NOCs in a cured meat model

New Insights into the Chemical Reactivity of Dry-Cured Fermented Sausages: Focus on Nitrosation, Nitrosylation and Oxidation

International audienceNitrite and nitrate are added to cured meat for their bacteriological, technological and sensorial properties. However, they are suspected to be involved in the formation of nitroso compounds (NOCs), such as potentially mutagenic nitrosamines, nitrosylheme and nitrosothiols. Controlling the sanitary and sensorial qualities of cured meat products by reducing these additives requires elucidating the mechanisms involved in the formation of NOCs. To this end, we studied the dose-response relationship of added sodium nitrite and/or sodium nitrate (0/0, 80/80, 0/200, and 120/120 ppm) on the formation of NOCs in dry cured fermented sausages. The results showed a basal heme iron nitrosylation in the absence of NaNO2/NaNO3 due to starter cultures. This reaction was promoted by the addition of NaNO2/NaNO3 in the other conditions. Reducing the dose to 80/80 ppm still limits lipid oxidation without the formation of non-volatile nitrosamines. Conversely, the addition of NO2/NO3 slightly increases protein oxidation through higher carbonyl content. The use of 80/80 ppm could be a means of reducing these additives in dry-cured fermented meat products

Detection of Frozen–Thawed Duck Fatty Liver by MALDI-TOF Mass Spectrometry: A Chemometrics Study

International audienceThe marketing of poultry livers is only authorized as fresh, frozen, or deep-frozen. The higher consumer demand for these products for a short period of time may lead to the marketing of frozen–thawed poultry livers: this constitutes fraud. The aim of this study was to design a method for distinguishing frozen–thawed livers from fresh livers. For this, the spectral fingerprint of liver proteins was acquired using Matrix-Assisted Laser Dissociation Ionization-Time-Of-Flight mass spectrometry. The spectra were analyzed using the chemometrics approach. First, principal component analysis studied the expected variability of commercial conditions before and after freezing–thawing. Then, the discriminant power of spectral fingerprint of liver proteins was assessed using supervised model generation. The combined approach of mass spectrometry and chemometrics successfully described the evolution of protein profile during storage time, before and after freezing-thawing, and successfully discriminated the fresh and frozen–thawed livers. These results are promising in terms of fraud detection, providing an opportunity for implementation of a reference method for agencies to fight fraud

A Proof of Concept to Bridge the Gap between Mass Spectrometry Imaging, Protein Identification and Relative Quantitation: MSI~LC-MS/MS-LF

International audienceMass spectrometry imaging (MSI) is a powerful tool to visualize the spatial distribution of molecules on a tissue section. The main limitation of MALDI-MSI of proteins is the lack of direct identification. Therefore, this study focuses on a MSI~LC-MS/MS-LF workflow to link the results from MALDI-MSI with potential peak identification and label-free quantitation, using only one tissue section. At first, we studied the impact of matrix deposition and laser ablation on protein extraction from the tissue section. Then, we did a back-correlation of the m/z of the proteins detected by MALDI-MSI to those identified by label-free quantitation. This allowed us to compare the label-free quantitation of proteins obtained in LC-MS/MS with the peak intensities observed in MALDI-MSI. We managed to link identification to nine peaks observed by MALDI-MSI. The results showed that the MSI~LC-MS/MS-LF workflow (i) allowed us to study a representative muscle proteome compared to a classical bottom-up workflow; and (ii) was sparsely impacted by matrix deposition and laser ablation. This workflow, performed as a proof-of-concept, suggests that a single tissue section can be used to perform MALDI-MSI and protein extraction, identification, and relative quantitation

Skeletal muscle lipid content and oxidative activity in relation to muscle fiber type in aging and metabolic syndrome

One of the most noticeable effects of aging is the reduction in skeletal muscle mass and strength (sarcopenia). The metabolic syndrome (MS) is also prevalent in old subjects, but its relevance to skeletal muscle characteristics has poorly been investigated. Immunohistochemical studies were performed with muscle biopsies from young (22 years) and old (73 years) men with and without MS to reveal age-dependent and MS-associated modifications of fiber-type characteristics. Atrophy of type II fibers and altered fiber shape characterized muscle aging in lean healthy men. In contrast, increased cross-sectional area of the most abundant type I and type IIA fibers, and reduced cytochrome c oxidase content in all fiber types, characterized MS. Aging and particularly MS were associated with accumulation of intramyocellular lipid droplets. Although lipids mostly accumulated in type I fibers, matrix-assisted laser desorption/ionization–mass spectrometry imaging of intramyocellular lipids did not distinguish fiber types, but clearly separated young, old, and MS subjects. In conclusion, our study suggests that MS in the elderly persons is associated with alterations in skeletal muscle at a fiber-type specific level. Overall, these fiber type-specific modifications may be important both for the age-related loss of muscle mass and strength and for the increased prevalence of MS in elderly subjects