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Construction De La Qualité Sensorielle Des Fromages De Type Cantal: Rôle

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Construction De La Qualité Sensorielle Des Fromages De Type Cantal: Rôle Université Blaise Pascal Université d’Auvergne N° D.U. 2776 ANNEE 2016 ECOLE DOCTORALE SCIENCE DE LA VIE, SANTE, AGRONOMIE, ENVIRONNEMENT N° D’ORDRE 708 Thèse Présentée à l’Université Blaise Pascal pour l’obtention du grade de DOCTEUR D’UNIVERSITE (SPECIALITE NUTRITION ET SCIENCES DES ALIMENTS) 14 décembre 2016 Marie Frétin Construction de la qualité sensorielle des fromages de type Cantal : rôle des interactions entre les communautés microbiennes et la composition de la matière grasse laitière des fromages Directeur de thèse : Bruno MARTIN Co-encadrantes : Céline DELBES et Anne FERLAY Président du jury Mr Télesphore SIME-NGANDO Directeur de recherche UMR CNRS 6023, Université Blaise Pascal Rapporteurs Mme Nathalie DESMASURES Professeur Université de Caen Normandie, Caen Mme Anne THIERRY Chargée de recherche INRA UMR 1253, Rennes Examinateurs Mr Pascal BONNARME Directeur de recherche INRA UMR 782, Thiverval-Grignon Mr Bruno MARTIN Ingénieur de recherche INRA UMR 1213, Theix Mme Céline DELBES Chargée de recherche INRA UR 545, Aurillac Unité de Recherches Fromagères (UR 545, URF, INRA) Unité Mixte de Recherches sur les Herbivores (UMR 1213, PERAQ, INRA) Résumé Résumé Construction de la qualité sensorielle des fromages de type Cantal : rôle des interactions entre les communautés microbiennes et la composition de la matière grasse laitière des fromages La qualité sensorielle des fromages au lait cru est variable selon la nature de l’alimentation des vaches laitières mais les mécanismes sous-jacents sont encore mal élucidés. Cette thèse avait pour objectif d’étudier les rôles respectifs et les interactions des composantes biochimique et microbiologique du lait, modulées par l’alimentation des vaches, dans la construction des caractéristiques sensorielles des fromages. Dans un premier temps, nous avons fait varier à la fois la composition biochimique et la composition microbiologique des laits en nous appuyant sur une expérimentation long terme comparant deux systèmes de pâturage qualifiés « d’extensif » (EXT) et de « semi-intensif » (SEMI). La structure des communautés bactériennes de la peau des trayons des vaches laitières, et dans une moindre mesure, celle des laits et des fromages de type Cantal ont varié selon les systèmes de production. Nos résultats confirment que le trayon est un réservoir potentiel de diversité microbienne, non seulement pour le microbiote du lait mais aussi pour celui du fromage. Les fromages EXT étaient caractérisés par une texture fondante et collante et une croûte moins épaisse comparativement aux fromages SEMI. Cependant, les deux systèmes de pâturage ont eu peu d’effet sur la flaveur des fromages, peut-être en raison de la trop grande similarité de la composition biochimique et microbiologique des laits des deux systèmes. Dans un deuxième temps, nous avons fait varier uniquement et de manière accentuée la composition de la matière grasse laitière via l’alimentation des vaches (herbe pâturée versus ensilage de maïs). Dans cette optique, des fromages de type Cantal ont été fabriqués à partir de deux crèmes pasteurisées de composition en acides gras différente et du même lait écrémé. Nous avons montré que l’effet de l’alimentation des vaches sur la texture des fromages était particulièrement lié à la composition de la matière grasse laitière tandis que celle-ci jouait un rôle mineur dans le développement de la flaveur des fromages. La crème la plus riche en acides gras saturés a été associée à une abondance relative plus élevée des ferments bactériens/fongiques et des OTUs dominants sur la surface des fromages et à la formation d’une croûte plus épaisse. Par comparaison, la croûte des fromages fabriqués avec une crème riche en acides gras insaturés était caractérisée par une plus grande diversité fongique et par la présence d’espèces sous- dominantes en abondance relative plus élevée. Cette thèse apporte des connaissances nouvelles sur l’effet de la composition de la matière grasse laitière sur les équilibres microbiens et le développement des caractéristiques sensorielles des fromages, et plus spécifiquement sur l’influence des deux composantes biochimique et microbiologique sur l’aspect de la croûte des fromages affinés. Mots-clés : assemblage du microbiote, matière grasse laitière, caractéristiques sensorielles des fromages, système de production laitier au pâturage, peau des trayons des vaches, métagénétique 3 Abstract Development of sensory quality of Cantal-type cheeses: effect of interaction between microbial communities and the composition of milk fat in cheeses The sensory quality of raw milk cheeses varies according to cow diet but the underlying mechanisms are still poorly understood. The aim of this PhD work was to understand the respective roles and possible interactions between the biochemical and microbiological milk components, modulated by cow diet, on the development of the sensory properties of cheeses. In the frame of a long term experiment comparing two groups of cows managed in two grazing systems qualified of “extensive” (EXT) or “semi-intensive” (SEMI), we made Cantal-type cheeses using milk varying both in its biochemical and microbial composition. The structure of the bacterial communities of the teat skin of dairy cows, and to a lesser extent, that of milk and cheese varied according to the production systems. Our results confirm that the teat skin is a potential reservoir of microbial diversity not only for the microbiota of milk but also for cheese. The EXT cheeses were characterized by a more melting and sticky texture and a thinner rind compared to the SEMI cheeses. However, due to the similarity of the biochemical and microbiological composition of the milks from both systems, the flavour of the resulting cheeses was little impacted. Therefore, we designed a second trial aiming at controlling the microbial composition of milk and accentuating the difference of milk fat composition, via cow diet (maize silage vs pasture). We made Cantal-type cheeses from two pasteurized creams with different fatty acid profiles added to the same skimmed milk. The milk fat composition had a strong influence on cheese texture but it played a minor role on the development of cheese flavour. The cream rich in saturated fatty acids was associated to a higher relative abundance of bacterial / fungal starter strains and of dominant OTUs on the surface of cheese, and to the development of a thicker rind. By comparison, the rind of cheeses made with the cream rich in unsaturated fatty acids was characterized by a higher fungal diversity and the presence of sub-dominant species in greater relative abundance. This thesis provides new knowledge on the effect of the composition of milk fat on the microbial balance and on the development of the sensory characteristics of cheeses, and more specifically on the influence of both biochemical and microbiological components of milk on the aspect of the rind of mature cheeses. Keys words: microbiota assembly, milk fat, cheese sensory properties, grazing based dairy system, cow teat skin, metabarcoding 5 Remerciements Remerciements Ce travail de thèse a été réalisé conjointement au sein de l’Unité de Recherches fromagères (UR 545, INRA Aurillac) et de l’équipe PERAQ (Pratiques d’Elevage, Robustesse, Adaptation et Qualité des produits, UMR 1213, INRA Theix). Je tiens à remercier la Région Auvergne pour la bourse régionale qui a permis de financer mes travaux de thèse. Un grand MERCI à mes trois encadrants Céline Delbès, Bruno Martin et Anne Ferlay pour m’avoir conseillé et guidé dans mes travaux, pour votre confiance et vos encouragements au cours de ces trois années, et votre investissement dans la correction du manuscrit et des articles. Céline, un merci particulier, pour ta disponibilité et ta gentillesse qui m’ont permis de relativiser dans les moments difficiles. Sincères remerciements à Anne Thierry et Nathalie Desmasures pour avoir accepté d’être rapporteures de cette thèse. Je vous remercie pour votre temps et vos remarques avisées. Je remercie également Télésphore Sime-Ngando pour avoir accepté de présider le jury et Pascal Bonnarme pour avoir accepté de faire partie de mon jury. J’adresse ensuite mes remerciements aux membres du comité de thèse pour leurs précieux conseils : Anne De La Foye (UMR 1213, Plate-forme d’exploration du métabolisme, INRA Theix), Erwan Engel (UR 370, QuaPA, INRA Theix), Claude Genot (UR 1268, BIA, INRA Nantes), Christophe Monnet (UMR 782, GMPA, INRA Grignon) et Anne Thierry (UMR 1253, STLO, INRA Rennes). Mes remerciements vont ensuite aux différentes personnes que j’ai pu côtoyer pendant ma thèse dans les différentes équipes et sans qui tous ces résultats n’auraient jamais pu voir le jour. Aux personnels des équipes AGL et Sybel (aujourd’hui PERAQ, UMR 1213, INRA Theix) pour toutes les analyses qui n’étaient pas de la microbio, votre convivialité, votre sympathie et votre bonne humeur lors de mon cours séjour à l’INRA de Theix. Merci à Emilie Tixier, Isabelle Constant, Yannick Faulconnier, Carole Delavaud, Didier Bany, Cyril Labonne, Anne Farruggia, Dominique Pomiès, Bertrand, Julien, Lehna et Jean-Baptiste. Aux personnels de l’Herbipôle (UE 1414, INRA Marcenat) pour la gestion des animaux sans lesquels cette thèse n’aurait jamais abouti. Merci à Philippe Pradel, Florence Fournier, Carole Cirié et Olivier Troquier pour m’avoir accompagné sur le terrain, au plus près des vaches, ce fut une expérience mémorable pour moi. Aux personnels de l’IUT d’Aurillac pour leur bienveillance et pour m’avoir permis de découvrir l’envers
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