Search

Le projet de recherche PROLIFIC (PROduits Laitiers et Ingrédients Fermentés Innovants pour des populations Cibles) va évaluer les bénéfices santé des produits laitiers fermentés à destination des jeunes enfants et des seniors.Le projet est porté par un consortium associant des industriels regroupés au sein de Bba Milk Valley et des équipes de recherche bretonnes et ligériennes.Son coût total est de 14 millions d'euros sur cinq ans, co-financé par les instituts de recherche, les industriels de Bba-Milk Valley et les Régions Bretagne et Pays de la Loire. PROLIFIC se décline en cinq thèses de doctorat, trois post-doctorats et 90 mois de CDD de niveau technicien à ingénieur. Yves Le Loir, directeur de l’UMR STLO, est chargé de coordonner le projet. Son lancement a eu lieu à la rentrée 2020.

The dairy platform of the STLO, Rennes, developed an innovative technology and more sustainable cheese making process (INRAE patent, From’Innov, WO2016108024). This technology consists of mixing a texture matrix obtained by ultrafiltration with one or more aromatic matrix. The latter is innovative: it allows obtaining a maximum of aroma in a short time, less than a few days. The principle is to grow each of microorganisms in an optimized middle with essential nutrients, optimal pH and temperature, and O2 if necessary. After adjusting the pH and temperature of the mix, we proceed with the coagulation then the molding, and the next day the demolding. The cheese is ready and contains the expected aromas. It needs five days to obtaining a soft-cheese with surface flora such as Penicillium and/or Geotrichum.We used this vector to define the consumer’s level of acceptability in view of a breakthrough technology.In this study, a commercial cheese (control product), representative of solubilized soft-cheeses, and two From'Innov cheeses (one close to control and the other more with more character) were studied. The acceptability and the main sensory properties of these three cheeses were evaluated by a panel of 142 consumers. The objective was to determine the hedonic appreciation of From'Innov cheeses, compared to that of commercial cheese, and to determine the sensory properties enhancing or penalizing the appreciation of different cheeses.The results showed that the hedonic appreciation of From’Innov cheese with character, although lower than that of commercial cheese, is satisfactory (5.6/10), despite a ripening time four times shorter. In addition, the intensity of the fruity notes and the character of this From’Innov cheese were judged to be close to ideal, whereas these two characteristics were judged to be insufficiently intense in commercial cheese. Finally, despite a lower salt (-20%) and fat (10%) content, the intensity of saltiness and fat of From’Innov cheese with character was judged to be close to ideal, very close to that of commercial cheese. The perspective of this study is to know whether our innovative technology will have a place in the current market and for which products, for whom?

For older adults (> 65 years old), undernutrition is a severe health problem that can impact the quality of life, induce or aggravate the development of diseases, and reduce life expectancy. Undernutrition can be defined as an inadequate intake of dietary energy and proteins combined with a low muscle mass. Diet-induced muscle mass and strength loss in older adults may be due to insufficient protein intake. Therefore, older adults need to increase the amount of high-quality ingested proteins, particularly foods rich in leucine, to promote muscle health. However, it is still unclear if changes in protein digestibility and absorption kinetics in old age may affect the anabolic effect of high-protein foods. The objective of this study was to investigate the in vitro digestion of two aged-tailored dairy products. A dairy dessert containing 10 % proteins and a spreadable cream cheese containing 24 % proteins were formulated with a ratio of whey proteins to caseins of 80 to 20 % (as opposed to milk, i.e., with a ratio of whey proteins to caseins of 20 to 80 %). This enrichment in whey proteins was used to increase the intake of branched amino acids, like leucine, to stimulate muscle protein synthesis in older adults. The rheological properties of these products were measured and related to their sensory properties (i.e., oral comfort) assessed by a panel of over 65 years-old subjects. Finally, the food matrix breakdown in the stomach and protein digestion have been studied with an in vitro digestion model adapted to the physiology of older adults. This new model results from an international consensus reached within the framework of the European project EAT4AGE in collaboration with the international research network INFOGEST. This study should improve our knowledge of the digestion of age-tailored dairy products in the aging gastrointestinal tract and help formulate innovative nutrient-dense food products that improve the bioavailability of proteins.

Sans risque, bonne pour la santé, pleine de saveurs, écologique... La fermentation, technique bien connue de nos ancêtres puis reléguée à certaines cultures avant de revenir en force depuis une vingtaine d'années, ne cesse de révéler ses bienfaits. Retour sur un engouement bien dans l'air du temps.

International audience; Eukaryotic cells are exposed to environmental and endogenous factors that induce DNA damage, thus affecting genomic integrity. The host cells counteract the consequences of lesions by DNA damage response and checkpoint systems that repair DNA structure or trigger cell death when DNA is irreparably damaged. S. aureus, a highly versatile gram-positive bacterium, can cause a multiplicity of human diseases ranging from mild superficial skin to life-threatening disseminated infections. S. aureus is one of the most prevalent pathogen that cause chronic ruminant mastitis that is very difficult to treat. Epithelial cells are able to sense microbes, creating an early line of defense against pathogens. Chronic S. aureus infection is likely to be associated with the internalization of the pathogen by host cells, where bacteria are protected from host defenses. Host cell cycle alteration is one of the highly sophisticated mechanisms pathogens use to hijack the main (defense) functions of the host cells, thus promoting their invasion and colonization. Recently we have shown S. aureus-induced cell cycle alteration in human and bovine epithelial cells.We aimed to investigate whether S. aureus can compromise host genomic integrity.We found that S. aureus can compromise host genomic integrity as indicated by bacteria-induced histone H2AX phosphorylation, a marker of DNA double strand breaks, in human epithelial HeLa and osteoblast-like MG-63 cells. This DNA damage is mediated by alpha phenol-soluble modulins (PSMα1–4), while a specific class of lipoproteins (Lpls), encoded on a pathogenicity island in S. aureus, dampens the H2AX phosphorylation thus counteracting the DNA damage. We demonstrated that this DNA damage is mediated by ROS (reactive oxygen species). DNA damage is followed by the induction of DNA repair that involves the ATM kinase-signaling pathway. An examination of S. aureus strains, isolated from the same patient during acute initial and recurrent bone and joint infections, showed that recurrent strains produce lower amounts of Lpls, induce stronger DNA-damage and prompt the G2/M transition delay to a greater extent that suggest an involvement of these mechanisms in adaptive processes of bacteria during chronicization. Our findings suggest that S. aureus infection has an impact on the genome and epigenome of host cells, which may exert patho-physiological dysfunctions and indicate that the balance between the levels of PSMα and Lpls expression impacts the persistence of the infection.

Early nutrition plays a dominant role in infant development and health. It is now understood that the infant diet impacts the gut microbiota and its relationship with gut function and brain development. However, its impact on the microbiota-gut-brain axis has not been studied in an integrative way. The objective here was to evaluate the effects of human milk (HM) or cow’s milk based infant formula (IF) on the relationships between gut microbiota and the collective host intestinal-brain axis. Eighteen 10-day-old Yucatan mini-piglets were fed with HM or IF. Intestinal and fecal microbiota composition, intestinal phenotypic parameters, and the expression of genes involved in several gut and brain functions were determined. Unidimensional analyses were performed, followed by multifactorial analyses to evaluate the relationships among all the variables across the microbiota-gut-brain axis. Compared to IF, HM decreased the α-diversity of colonic and fecal microbiota and modified their composition. Piglets fed HM had a significantly higher ileal and colonic paracellular permeability assessed by ex vivo analysis, a lower expression of genes encoding tight junction proteins, and a higher expression of genes encoding pro-inflammatory and anti-inflammatory immune activity. In addition, the expression of genes involved in endocrine function, tryptophan metabolism and nutrient transport was modified mostly in the colon. These diet-induced intestinal modifications were associated with changes in the brain tissue expression of genes encoding the blood-brain barrier, endocrine function and short chain fatty acid receptors, mostly in hypothalamic and striatal areas. The integrative approach underlined specific groups of bacteria (Veillonellaceae, Enterobacteriaceae, Lachnospiraceae, Rikenellaceae, and Prevotellaceae) associated with changes in the gut-brain axis. There is a clear influence of the infant diet, even over a short dietary intervention period, on establishment of the microbiota-gut-brain axis.

International audience; ContextGut inflammation constitutes a growing health concern in developed countries. It may consist in spontaneous ailments of the gut, involving both the host immune system and microbiota, such as IBD, including ulcerative colitis and Crohn’s disease. It may be caused by a medical treatment, such as mucositis induced by cancer chemotherapy and/or radiotherapy. It coincides with a dysbiosis including a lack of anti-inflammatory bacteria. As an example, propionibacteria are lacking in the microbiota of newborns that develop necrotizing enterocolitis.Methods and resultsWe thus focused on the immunomodulatory properties of GRAS propionibacteria. Selected strains of Propionibacterium freudenreichii induced the regulatory IL-10 cytokine in human immune cells (Foligné et al., 2010, 2013), depending on surface proteins (Le Marechal et al., 2015). Mutation of the slpB gene suppressed this immunomodulatory effect and the resulting slpB mutant induced a rather proinflammatory response (Deutsch et al., 2017). Consumption of wild-type P. freudenreichii protected from colitis induced by both TNBS and by DSS. It alleviated severity of symptoms, modulated local and systemic inflammation, as well as colonic oxidative stress and epithelial cell damages (Plé et al., 2015, 2016; Rabah et al., 2020). Accordingly, consumption of Lactococcus lactis NCDO 2118 harboring pXIES-SEC:slpB and expressing the propionibacterial SlpB reduced severity of colitis, lowered weight loss, disease activity index, shortening of the colon length, and histopathological score, compared with mice treated with L. lactis NCDO 2118 wild-type strain.In the context of mucositis induced by the chemotherapy 5-FU, P. freudenreichii prevented weight loss, reduced inflammation and consequently intestinal damages. It regulated key markers, including Claudin-1 and IL-17a genes, as well as IL-12 and IL-1β cytokines levels (Cordeiro et al., 2018). Mutant strain slpB displayed opposite regulatory effect on cld1 expression and on IL-12 levels, and failed to afford protection towards 5-FU-mucositis (do Carmo et al., 2019).ConclusionThis work emphasizes the importance of SlpB in P. freudenreichii ability to reduce both mucositis and colitis inflammation. It opens perspectives for the development of probiotic products aimed at decreasing side effects of chemotherapy and at helping treatment of colitis, thanks to GRAS bacteria.ReferencesCordeiro, B. F., Oliveira, E. R., Silva, D., H, S., Savassi, B. M., Acurcio, L. B., et al. (2018). Whey Protein Isolate-Supplemented Beverage, Fermented by Lactobacillus casei BL23 and Propionibacterium freudenreichii 138, in the Prevention of Mucositis in Mice. Front. Microbiol. 9. doi:10.3389/fmicb.2018.02035.Deutsch, S.-M., Mariadassou, M., Nicolas, P., Parayre, S., Le Guellec, R., Chuat, V., et al. (2017). Identification of proteins involved in the anti-inflammatory properties of Propionibacterium freudenreichii by means of a multi-strain study. Sci Rep 7, 46409. doi:10.1038/srep46409.do Carmo, F. L. R., Rabah, H., Cordeiro, B. F., da Silva, S. H., Pessoa, R. M., Fernandes, S. O. A., et al. (2019). Probiotic Propionibacterium freudenreichii requires SlpB protein to mitigate mucositis induced by chemotherapy. Oncotarget 10, 7198–7219. doi:10.18632/oncotarget.27319.Foligné, B., Breton, J., Mater, D., and Jan, G. (2013). Tracking the microbiome functionality: focus on Propionibacterium species. Gut 62, 1227–1228.Foligné, B., Deutsch, S. M., Breton, J., Cousin, F. J., Dewulf, J., Samson, M., et al. (2010). Promising immunomodulatory effects of selected strains of dairy propionibacteria as evidenced in vitro and in vivo. Appl.Environ.Microbiol. 76, 8259–8264.Le Marechal, C., Peton, V., Ple, C., Vroland, C., Jardin, J., Briard-Bion, V., et al. (2015). Surface proteins of Propionibacterium freudenreichii are involved in its anti-inflammatory properties. J.Proteomics. 113C, 447–461.Plé, C., Breton, J., Richoux, R., Nurdin, M., Deutsch, S.-M., Falentin, H., et al. (2016). Combining selected immunomodulatory Propionibacterium freudenreichii and Lactobacillus delbrueckii strains: Reverse engineering development of an anti-inflammatory cheese. Mol Nutr Food Res 60, 935–948. doi:10.1002/mnfr.201500580.Plé, C., Richoux, R., Jardin, J., Nurdin, M., Briard-Bion, V., Parayre, S., et al. (2015). Single-strain starter experimental cheese reveals anti-inflammatory effect of Propionibacterium freudenreichii CIRM BIA 129 in TNBS-colitis model. Journal of Functional Foods 18, 575–585. doi:10.1016/j.jff.2015.08.015.Rabah, H., do Carmo, F. L. R., Carvalho, R. D. de O., Cordeiro, B. F., da Silva, S. H., Oliveira, E. R., et al. (2020). Beneficial Propionibacteria within a Probiotic Emmental Cheese: Impact on Dextran Sodium Sulphate-Induced Colitis in Mice. Microorganisms 8, 380. doi:10.3390/microorganisms8030380.- PF induces IL-10 in human PBMCs- Surface layer extraction reduces IL-10 induction- Extracted surface layer induces IL-10- Mutation of the SlpB gene reduces IL-10 induction- PF prevents colitis- PF prevents mucositis- PF slpB mutant does not prevent mucositis- Lactococcus lactis expressing SlpB prevents colitis

This conference is a major event in the field of Food, Nutrition and Health. It is organized in the frame of the INFOGEST research network(www.cost-infogest.eu) whose objective is to “improve the health properties of food by sharing our knowledge on the digestive process”. INFOGESTis a global network of approximately 582 research scientists (academic and food companies) from 150 institutions across 46 countries.; International audience; Infant formulas (IFs), the only adequate substitute to breastmilk, are complex matrices that require numerous ingredients and processing steps. The objective was to understand how protein structure within IFs modulates their digestive kinetics. Four IFs (A/B/C/D), containing whey protein (WP) ingredients with different denaturation and glycation levels (A/B/C) and caseins with different structures (C/D), were subjected to infantin vitro dynamic digestion (DIDGI®). Digesta were regularly sampled to follow structural changes (A4F, microscopy), proteolysis (OPA, LC-MS-MS) and lipolysis (GC-MS). Data were analysed using repeated measures ANOVA. Before digestion, lipoprotein structures were different among IFs. IF-A, characterized by higher glycated and denatured WP rates, tended to be more digested at 180min of intestinal phase than IF-C/D (degree of proteolysis +16% and lipolysis +5.2%). Peptides (protein-origin independent) appeared sooner into the gastric phase for IF-D (at 80 min vs. 180 min for IF-A/B), suggesting that the initial bigger lipoprotein structures in the matrix were less dense and more accessible to pepsin. Different bioactive peptides kinetics were also observed among IFs during digestion. Overall, it highlights the importance of the structure of protein ingredients (WPs and caseins)selected for IFs. Further investigation will be conducted in vivo (mini-piglets) to complete these data.

International audience; La modélisation et l’optimisation des procédés agroalimentaires est une tâche difficile dû à la complexité des produits agroalimentaires, le manque de connaissances sur les mécanismes limitant les performances [1] et l’hétérogénéité des variables impliquées. C’est le cas pour la microfiltration tangentielle 0.1 μm de lait écrémé (MF 0.1 μm). Cette opération est généralement utilisée dans l’industrie laitière pour séparer les protéines : micelles de caséines natives (rétentat) utilisées dans les fromages et les protéines de lactosérum (perméat) principalement utilisées dans les formulations alimentaires à destination des populations spécifiques (personnes âgées, nourrissons, sportifs). En dépit du fort intérêt porté à la MF 0.1 μm dans le secteur de l’industrie agroalimentaire, ce procédé de transformation n’a pas été intégralement optimisé au regard des objectifs des utilisateurs. Les choix de conception, dimensionnement et conditions opératoires sont principalement basées sur le savoir-faire des équipementiers et des connaissances expertes disponibles. Dans la littérature, l’optimisation de la MF 0.1 μm de lait écrémé est abordée en mono-objectif sur un problème posé empiriquement [2] ou sur l’influence spécifique d’une variable d’intérêt sur un groupe de variables choisies [3]. Ce travail vise à optimiser la microfiltration tangentielle 0.1 μm de lait écrémé depuis la conception jusqu’aux performances des produits avec des objectifs d’optimisation contradictoires en intégrant des connaissances expertes et scientifiques dans la formulation d’un problème multiobjectif. La méthodologie, basée sur « l'intégration des connaissances » se compose de quatre étapes : 1) l'acquisition de connaissances scientifiques et expertes sur les objectifs à optimiser ; 2) la modélisation des objectifs sous forme de fonctions calculables ; 3) la résolution du problème multiobjectif en utilisant un algorithme d'optimisation métaheuristique adapté ; 4) l'aide à la décision multicritère. Les objectifs considérés sont de maximiser la composition des fractions rétentat et perméat, de maximiser le rendement de récupération des protéines dans la fraction perméat et de minimiser les coûts économiques. Environ 1000 solutions Pareto-optimales ont été trouvées, dont des solutions proches de celles utilisées dans l'industrie et à moindre coût. L'optimisation multiobjectif de la MF a ouvert de nouvelles réflexions sur la conception des installations et les combinaisons de conditions opératoires en améliorant les caractéristiques du produit tout en maintenant constants les coûts d'investissement et de production. Le résultat de l'optimisation est une preuve de concept dont la faisabilité et la viabilité à l'échelle industrielle doivent être validées. Ces travaux ouvrent de nouvelles perspectives pour l'optimisation multiobjectif des procédés alimentaires.

Background Streptococcus thermophilus is a Gram-positive bacterium widely used as starter in the dairy industry as well as in many traditional fermented products. In addition to its technological importance, it has also gained interest in recent years as beneficial bacterium due to human health-promoting functionalities. The objective of this study was to inventory the main health-promoting properties of S. thermophilus and to study their intra-species diversity at the genomic and genetic level within a collection of representative strains. Results In this study various health-related functions were analyzed at the genome level from 79 genome sequences of strains isolated over a long time period from diverse products and different geographic locations. While some functions are widely conserved among isolates (e.g., degradation of lactose, folate production) suggesting their central physiological and ecological role for the species, others including the tagatose-6-phosphate pathway involved in the catabolism of galactose, and the production of bioactive peptides and gamma-aminobutyric acid are strain-specific. Most of these strain-specific health-promoting properties seems to have been acquired via horizontal gene transfer events. The genetic basis for the phenotypic diversity between strains for some health related traits have also been investigated. For instance, substitutions in the galK promoter region correlate with the ability of some strains to catabolize galactose via the Leloir pathway. Finally, the low occurrence in S. thermophilus genomes of genes coding for biogenic amine production and antibiotic resistance is also a contributing factor to its safety status. Conclusions The natural intra-species diversity of S. thermophilus, therefore, represents an interesting source for innovation in the field of fermented products enriched for healthy components that can be exploited to improve human health. A better knowledge of the health-promoting properties and their genomic and genetic diversity within the species may facilitate the selection and application of strains for specific biotechnological and human health-promoting purpose. Moreover, by pointing out that a substantial part of its functional potential still defies us, our work opens the way to uncover additional health-related functions through the intra-species diversity exploration of S. thermophilus by comparative genomics approaches.

International audience; Decentralized food production, usage of high-quality, local raw material, compliance with traditional processing methods, and preservation of nutritional value are attributes of Intermediate Food Value Chains (ivc) and may significantly contribute to the agri-food sustainability transformation. As these practices typically come with an extra cost, the consumer should be aware and convinced of them so that a premium price can be paid. But the food chain is complex and such a proof is challenging: Raw material come from multiple sources, it undergoes complex processing,while the package-imprinted attributes presently display mainly nutritional attributes. The large number of intermediates in the food supply chain adds to the complexity and fraud possibilities, while each production line has its own characteristics. This creates a challenge in terms of transparency and trust, from the consumer perspective. Blockchain as an open, distributed ledger can record transactions in a verifiable and permanent way and it promises transparency in food chains [1]. In this work, we investigate whether Blockchain can be sustainably applied in the dairy ivc of H2020 FAIRCHAIN project, identifying four challenges and our working practices. The ivc focuses on Feta cheese production; Feta is the most known, traditional Greek product of designated origin (PDO). It is based only on sheep and goat milk, under defined percentages, processed in specific regions, with requirements of minimum maturation periods. The information offered to the consumer covers the nutritional aspects and is difficult to verify. Challenge #1, which data to register, so that they are quantitative and meaningful? Through the FAIRCHAIN co-creation process we have identified a set of high-level attributes (taste, origin, integrity) and associated them with quantitative operational parameters in the end-to-end process, such as milk temperatures and pH, microbiological checks, milk mixing and percentages, maturation duration, IoT readings. These carefully selected attributes create the ‘digital product passport’. Challenge #2, how to guarantee the correctness of the data? If erroneous data are inserted at the edge, the Blockchain cannot correct them. While this risk cannot be eliminated, it can be mitigated with data verification rules (such as acceptable data ranges of input data), regular checks, and anomaly detection [2-4]. Challenge #3, how to rationalize Blockchain transaction fees and allow for verification on behalf of the consumer? Not all info can be registered in the Blockchain due to the need for performance and the transaction fees. While technological choices (such as the consensus algorithms) can affect the performance, the data granularity is an open question. Anchoring mechanisms (from private to public Blockchains). Data verification is based on (Ethereum) smart contracts and visualized through simple mobile app (confronting digital divide aspects as presented in [5]).Challenge #4, how to facilitate the involvement of stakeholders and build an application ecosystem? The offering of open APIs (Application Programming Interfaces) paves the way for an ecosystem of applications involving externalstakeholders (such as public authorities for food certifications). The platform is currently reaching a stable version and it is undergoing evaluation for identifying adoption challenges and quantifying the operational overhead

International audience; Purpose Numerous studies, including our previous work with lemon juice, have reported that low-pH meals reduce the glycemic response to starchy foods. However, the underlying mechanism is not yet understood. Tea, for its polyphenol content, has also been investigated. The main objective of this research was to concurrently study gastric emptying, appetite perceptions and glycemic responses to bread consumed with water, tea, or lemon juice. Methods In this randomized, crossover intervention, ten participants consumed equal portions of bread (100 g) with 250 mL of water, water-diluted lemon juice, or black tea at breakfast. Gastric volumes, blood glucose concentrations and appetite perceptions were alternately assessed over 180 min using magnetic resonance imaging, the finger-prick method and visual analogue scales, respectively. Results Compared to water, lemon juice led to a 1.5 fold increase of the volume of gastric contents, 30 min after the meal (454.0 ± 18.6 vs. 298.4 ± 19.5 mL, x ± SEM P < 0.00001). Gastric emptying was also 1.5 times faster (P < 0.01). Conversely, lemon juice elicited a lower glycemic response than water (blood glucose concentrations at t = 55 min were 35% lower, P = 0.039). Tea had no effect. Changes in appetite perceptions and gastric volumes correlated well, but with no significant differences between the meals. Conclusions Lemon juice lowered the glycemic response and increased both gastric secretions and emptying rate. The results are compatible with the hypothesis that the reduction of the glycemic response is mainly due to the interruption of starch hydrolysis via the acid-inhibition of salivary α-amylase.

International audience; IntroductionGut inflammation constitutes a growing health concern in developed countries. It may consist in spontaneous ailments of the gut, involving both the host immune system and microbiota, such as IBD, including ulcerative colitis and Crohn’s disease. It may be caused by a medical treatment, such as mucositis induced by cancer chemotherapy and/or radiotherapy. It coincides with a dysbiosis including a lack of anti-inflammatory bacteria. As an example, propionibacteria are lacking in the microbiota of newborns that develop necrotizing enterocolitis.Methods Dairy propionibacteria strains were tested in vitro on human PBMCs with respect to their ability to induce immunomodulatory cytokines. Selected strains were then tested in vitro with respect to their ability to protect mice from TNBS-induced colitis, from DSS-induced colitis, and from 5FU-induced mucositis.ResultsWe thus focused on the immunomodulatory properties of GRAS propionibacteria. Selected strains of Propionibacterium freudenreichii induced the regulatory IL-10 cytokine in human immune cells (Foligné et al., 2010, 2013), depending on surface proteins (Le Marechal et al., 2015). Mutation of the slpB gene suppressed this immunomodulatory effect and the resulting slpB mutant induced a rather proinflammatory response (Deutsch et al., 2017). Consumption of wild-type P. freudenreichii protected from colitis induced by both TNBS and by DSS. It alleviated severity of symptoms, modulated local and systemic inflammation, as well as colonic oxidative stress and epithelial cell damages (Plé et al., 2015, 2016; Rabah et al., 2020). Accordingly, consumption of Lactococcus lactis NCDO 2118 harboring pXIES-SEC:slpB and expressing the propionibacterial SlpB reduced severity of colitis, lowered weight loss, disease activity index, shortening of the colon length, and histopathological score, compared with mice treated with L. lactis NCDO 2118 wild-type strain (Belo et al., 2021).In the context of mucositis induced by the chemotherapy 5-FU, P. freudenreichii prevented weight loss, reduced inflammation and consequently intestinal damages. It regulated key markers, including Claudin-1 and IL-17a genes, as well as IL-12 and IL-1β cytokines levels (Cordeiro et al., 2018). Mutant strain slpB displayed opposite regulatory effect on cld1 expression and on IL-12 levels, and failed to afford protection towards 5-FU-mucositis (do Carmo et al., 2019).P. freudenreichii was further shown to produce extracellular vesicles (EVs), which mimic the immunomodulatory features of propionibacteria in vitro by modulating NF-κB transcription factor activity and IL-8 release in cultured human intestinal epithelial cells (Rodovalho et al., 2020). Proteomic analysis revealed presence of SlpB in these EVs.DiscussionThis work emphasizes the importance of SlpB in P. freudenreichii ability to reduce both mucositis and colitis inflammation. It opens perspectives for the development of probiotic products aimed at decreasing side effects of chemotherapy and at helping treatment of colitis, thanks to GRAS bacteria.

Innovative agri-food value chains : boosting sustainability-oriented competitiveness under the programme SC 2 “Food security, sustainable agriculture and forestry, marine, maritime and inland water research and the bioeconomy”

Le lait n’est pas un aliment comme les autres. Consommé depuis le Néolithique et la domestication des ruminants, il est décliné en mille et un produits différents, est intégré à mille et une recettes de cuisine, et suscite des passions et des débats agités plus que jamais d’actualité… Aliment exclusif du nourrisson, sa place dans la diète à l’âge adulte dépend des cultures. Inépuisable source d’innovations, c’est un objet d’étude fascinant par sa complexité biochimique et nutritionnelle, optimisée par les mammifères… depuis 310 millions d’années.Par un jeu de questions-réponses, les auteurs donnent les clés de toutes les facettes du lait et des produits laitiers en termes de nutrition, santé, économie, symbolique, histoire, culture…À travers 50 questions des plus insolites — Les hommes peuvent-ils allaiter ? ou Peut-on faire de l’alcool avec du lait ? —, des plus sérieuses — Y a-t-il des utilisations non alimentaires du lait ? Est-ce « durable » de consommer des produits laitiers ? Tous les laits sont-ils fromageables ? — aux plus anecdotiques — C’est quoi les dents de lait ? Le lait de jabot : vrai ou faux lait ? —, vous comprendrez qu’il reste encore beaucoup à découvrir sur les effets du lait (fermenté ou non) sur la santé, sa composition, ses impacts sur l’environnement, sa transformation…De quoi animer vos repas de famille… autour du plateau de fromages !

Dans le cadre de la valorisation des ressources microbiennes, le département MICA a souvent été confronté à la gestion de la propriété intellectuelle associée à ces ressources et aux questions réglementaires liées à l’Accès et au Partage des Avantages découlant de leur utilisation (Protocole de Nagoya/APA). Pour répondre à ces problématiques, le département MICA a constitué un groupe de travail coordonné par Angèle Charrier, Chargée de Partenariat et d’Innovation du département et impliquant des membres du réseau CIRM et d’INRAE transfert. L’objectif de ce groupe de travail était de concevoir un outil destiné aux chercheurs utilisateurs de micro-organismes. Il a abouti à la création du mémo « Bonnes pratiques en valorisation demicro-organismes ». La rédaction de ce mémo s’est appuyée sur les particularités des micro-organismes en général. Les questions relatives à l’origine des ressources (respect du protocole de Nagoya et de l’APA), leur historique d’acquisition et de diffusion (INRAE est-il le détenteur légitime de ces ressources ? La diffusion de ces ressources est-elle maîtrisée ?) et les conditions d’utilisation et de valorisation (qu’ai-je le droit de faire avec ces ressources et sont-elles valorisables ?) restent valables pour tous types de ressources biologiques. Dans ce contexte, une démarche semblable pourrait être envisagée pour des ressources biologiques, autres que microbiennes, soumises aux mêmes contraintes de propriété intellectuelle et réglementaires mais portant d’autres spécificités. Ce mémo, qui a pris la forme de la plaquette présentée ici, a pour ambition d’aider les chercheurs à respecter la propriété intellectuelle et la réglementation en vigueur, dans le cadre d’un projet de recherche ou de valorisation

Numerous studies have reported a 20-50% reduction of the glycaemic response to starch-rich meals by low-pH drinks or foods. Although different possible explanations have been put forward, our work using bread and lemon juice (pH≈2.3) as a case study has pointed to the acid-induced inhibition of salivary α-amylase as the most likely underlying mechanism. Therefore, our aim was to study the digestion of a different subset of such meals to further investigate this hypothesis. Four meals have been replicated in the lab both with and without their pH-lowering ingredient. Their initial pHs were measured and they were then digested in vitro using the semi-dynamic INFOGEST protocol. Each digestion included oral, gastric and intestinal phases and experiments were conducted in triplicate. Samples were collected at different time-points to monitor the extent of starch hydrolysis and the MW weight distributions of the polysaccharide hydrolysates throughout the experiments. Low pH meals exhibited a marked reduction of the proportion of starch digested compared to their neutral-pH counterparts. The agreement between these results with those of previous human studies confirms our earlier observations and offers new perspectives for the development of strategies to improve the glycaemic responses to starch-rich foods.

Introduction Intestinal epithelial barrier (IEB) integrity plays a central role in the pathogenesis of inflammatory bowel disease (IBD). There is a growing interest in dietary compounds that can strengthen IEB. First, candidate strains of dairy propionic bacteria such as Propionibacterium freudenreichii are able to reduce inflammation and improve barrier strength. Knowing that certain products of omega-6 polyunsaturated fatty acid (n-6 PUFA) metabolism also reinforce IEB, and that bacteria can modulate the bioavailability of host lipid compounds, this project aims to identify strains of P. freudenreichii that could protect against colitis by strengthening IEB and increasing the bioavailability of certain n-6 PUFA. MethodsA screening of thirty-three strains of P. freudenreichii was performed based on the following criteria: 1) characterization of the ability to induce production of anti-inflammatory cytokines (IL-10) by stimulation of human PBMCs 2) identification of surface proteins (S-layer protein) responsible for anti-inflammatory properties 3) measurement of their ability to increase the bioavailability of certain n-6 PUFAs, such as conjugated linoleic acids (CLAs). The ability of the selected fermented products to prevent the inflammation-induced increase in permeability of an IEB was then studied. ResultsAll strains are able to induce IL-10 production and release CLAs. Ten strains show an anti-inflammatory S-layer profile. Some fermented products significantly prevent the inflammation-induced increase in permeability, and may involve surface proteins and bioactive lipids. DiscussionThese results will allow the selection of strains with anti-inflammatory as well as pro-repair potential to be tested in vivo to reduce colitis, with or without n-6 PUFA supplementation.

compréhension des interactions entre protéines végétales et protéines de blanc d’œuf pour permettre l’amélioration de la « techno-fonctionnalité » des mélanges, notamment pour la stabilisation des mousses et émulsions, l’optimisation de ces fonctionnalités et l’identification des conditions permettant de limiter leur sensibilité aux traitements thermiques.

Background Saliva may play a role in digestion through its enzymatic content or physico-chemical impact on the food bolus or chyme. Salivary proteins also interact with tannins, which could counteract their effect on digestive enzymes. We here evaluated the impact of saliva on gastric proteolysis of solid or liquid dairy products, including one contaning tannins.MethodSwiss-type cheese (C), milk in water (M) or milk in black tea (T), all in presence of human saliva vs water, were digested using semi-dynamic (C) and static conditions (M,T), respectively. Proteolysis was monitored throughout gastric digestion by OPA assay in all samples. The degree of hydrolysis was also calculated for cheese using acid titration data.ResultsFor cheese, the presence of saliva delayed slightly the initiation of proteolysis. OPA measurements suggested a small proteolytic inhibition by saliva. For milk, the presence of tannins increased gastric proteolysis (possibly by denaturing the pepsin substrates) but saliva had no substantial impact on OPA measurements, whether the matrix contained tannins or not.Conclusion In the conditions used, saliva had a moderate, if any, impact on gastric digestion of dairy products. Due to the documented effect of saliva on emulsions stability, the study could be extended to lipolysis.

Le CBL (Club des Bactéries Lactiques) est une manifestation scientifique qui réunit chercheurs, enseignants-chercheurs et industriels R&D, pour échanger sur les avancées scientifiques et techniques réalisées dans le domaine des bactéries lactiques. Les thèmes abordés sont le reflet des larges potentiels de ces bactéries et de leurs applications : fermentation alimentaire, santé humaine et animale, probiotique, environnement…; International audience; En fabrication fromagère, les bactéries lactiques (BL) et propioniques sont des actrices clés de la production de métabolites conférant les qualités nutritionnelles et organoleptiques aux fromages. Pourtant, la contribution de chaque espèce à la qualité finale du fromage n’est pas totalement élucidée. L’objectif était de déterminer quelles espèces contribuent à acidifier et produire des composés d’arôme, par quelles voies métaboliques, selon quelle temporalité et aussi d’investiguer les interactions métaboliques bactériennes contribuant au fonctionnement de l’écosystème fromager.Nous avons séquencé et annoté : Lactococcus lactis subsp. lactis biovar diacetylactis CIRM-BIA1206 (LL), Lactiplantibacillus plantarum CIRM-BIA465 (LP), Propionibacterium freudenreichii CIRM-BIA122 (PF). Nous avons reconstruit les voies métaboliques et élaboré un modèle métabolique de communauté. Un fromage à pâte pressée non cuite sans croûte a été réalisé avec ces bactéries et analysé pendant la fabrication et l’affinage (4 réplicats). Des dénombrements bactériens, des dosages de sucres, d’acides organiques et de composés d’arôme [1] ont été réalisés ainsi qu’un séquençage des ARN bactériens. L’analyse des gènes différentiellement exprimés a permis de montrer à quel moment de la fabrication étaient induits les gènes impliqués dans le catabolisme du lactose et du citrate et dans les voies de synthèse de différents composés d’arômes : acides lactique, acétique, propionique, isovalérique (arôme ‘vieux fromage’), diacétyle (arôme beurré). Le catabolisme du lactose était opéré chez LL via la voie du tagatose pendant l’acidification puis via la voie de Leloir et uniquement par cette dernière voie chez LP. La production de diacétyle était effectuée par LL dès le début de la fabrication puis plus modérément par LP. Les synthèses d’acides propionique et isovalérique ont été attribuées à PF et les métabolismes correspondants induits pendant la phase d’affinage. Enfin, les voies de fermentation mixtes ayant été induites pendant l’affinage, l’acide acétique a été vraisemblablement produit par les trois espèces à cette étape. L’implémentation du modèle métabolique communautaire dans l’outil Smetana [2] a révélé les bases moléculaires du commensalisme précédemment évoqué entre BL et PF [3–5]. Les BL produisent de l’acide lactique et potentiellement du glycérol, de la sérine et de la phénylalanine au profit de PF. Ces interactions identifiées in silico restent à valider in vitro.L’ensemble de ces résultats font de la métatranscriptomique associée aux modèles métaboliques, des outils de choix pour mieux comprendre et maîtriser les métabolismes et les interactions régissant le fonctionnement des écosystèmes fromagers.

The adage “in unity is strength” also applies to fermented foods: the association of lactic acid bacteria (LAB) strains brings benefits to their organoleptic properties and potential health benefits. Such functionalities can depend on positive interactions, as observed between LAB in yogurt. Finding waysto create new functional associations of LAB strains is challenging, especially in the context of the food transition where new resources, notably proteins, are increasingly used. The objective of this PhD work was to understand how to promote positive interactions between LAB, on the basis of their nitrogen metabolism, to conceive new food applications. The interactions were studied in cocultures that associated a proteolytic (prot+) and a non-proteolytic (prot-) strain. These prot+/prot- pairs were first grown in a model medium containing milk and lupin proteins as sole nitrogen nutriments, in which the growth of prot- strain thus depended on the peptides and amino acids released by the prot+ strain. Strong,weak, or no positive interactions were observed between prot+ and prot- strains. Peptidomics results showed that the release of branched-chain amino acids either free or within peptides was important in the positive interactions observed. We then studied the impact of these interactions in alternative yogurts made from milk and lupin, characterized for their physical and sensory properties. Positive interactionsbetween LAB led to diversified functionalities of alternative yogurts, such as flavour compound and organic acid formation, and texture improvement. This work brings new knowledge on the interactions between LAB, which is useful in designing future fermented foods.

Aim: Cheese whey is the watery part obtained after milk curdling in cheese-making. Because it contains high quality proteins, whey is mainly valorized by industrial processed to obtain value-added food ingredients. However, these processes are only applicable to some types of whey, are energy-consuming and only concern large-scale dairy plants. As part of FAIRCHAIN project, we aimed at developing whey-based beverages using fermentation as alternative whey valorization process. This study explores the ability of lactic acid bacteria (LAB) to ferment different types of whey, to obtain the best aroma profiles for the beverages.Method: Two types of whey were assessed: a sweet whey from Morbier cheese and an acid one from Comté cheese. 125 LAB strains belonging to 27 species, were selected for their capacity to utilize lactose and their GRAS status. They were evaluated for their ability to ferment both wheys. After fermentation, the products were assessed for their acidification and sensory properties (tastes and sniffing). Sugar content and volatile profile of fermented wheys were evaluated by HPLC and GC-MS respectively.Results: Fermentation outcomes differed according to the whey type: 34 strains did not acidify sweet whey, while 91 strains did not acidify the acid one. For a given whey, acidification was species-dependent: Lactobacillus helveticus and Lactobacillus delbrueckii strains showed the best acidification capacities in both wheys although acidification was higher on sweet whey compared to the acid one. Sensory profile analysis led to different results and were strain-dependent. Regarding sweet whey, 40 strains (12.5%) encompassing 12 species showed a promising aroma profile while only 20 belonging to 7 species were retained for acid whey. Both criteria considered, 26% strains tested on sweet whey showed a good acidification ability and an aroma profile deemed to be acceptable, whereas only 20 strains were retained for Comté acid whey. Noteworthy, only 6 strains showed promising results on both wheys.Conclusion: This study highlighted the huge diversity of the metabolic profiles of LAB strains when used for fermentation of different whey types. Fermentation by well selected bacteria LAB thus appears as a sustainable and an inexpensive process, to valorize whey.

Fermentation of vegetables has recently received a renewed interest in the USA and in Western Europe, where they are little consumed, in contrast with central Europe and Asia. FLEGME is a citizen science project focused on fermented vegetables, which gathers various actors including citizens, SMEs, agricultural schools, culinary journalists, and researchers. One of its goal is to characterize the microbial communities and the safety of home-made fermented vegetables. A set of 75 fermented vegetables was collected from citizens that manufacture such products for their personal consumption. Several microbial groups were analysed by culture-dependent methods, including lactic acid bacteria (LAB), yeasts, and some pathogenic bacteria. pH was also measured. 1 to 3 isolates were randomly picked up from LAB and yeast plates and identified by 16S and D1/D2 sequencing. Samples were also analysed by metabarcoding analysis. The received included 23 types of vegetables, mainly cabbage (27%), followed by carrots (19%) and beets (12%). Their age ranged from 2 weeks to 4 years. Their median pH was 3.6, only 2 samples had a pH over 4.5. LAB represented the dominant population. Their median concentration was 7.5x104 CFU/g, but varied from non-detectable values to 6x108 CFU/g. No significant relationship was found between LAB counts and the type of vegetable. However, LAB counts significant depended on the age of samples, with high LAB counts most frequently observed in the youngest samples (< 100 days). Yeasts were detected in nearly half the samples, with counts ranging from 102 to 9x107 CFU/g. Enterococci were detected in only 5 samples at counts < 105 CFU/g. No pathogenic bacteria were detected. 90 LAB clones and 47 yeast clones were isolated, belonging to 31 species, the most common being Levilactobacillus brevis and Lactiplantibacillus plantarum/paraplantarum (21 and 20 % of total bacterial isolates, respectively) and Saccharomyces cerevisiae, Debaryomyces hansenii, and Wickerhamomyces anomalus for yeasts. Metagenomic results showed that Lactobacillacae was the most represented family, with Lactiplantibacillus as most frequent genus encountered), followed by Streptococcaceae, Leuconostocaceae and Staphylococcaceae, with Lactococcus, Weissella, and Staphylococcus, respectively, as most frequent genera.

Propionibacterium freudenreichii is a probiotic Gram-positive bacterium with promising immunomodulatory properties. It modulates regulatory cytokines, mitigates the inflammatory response in vitro and in vivo These properties were initially attributed to specific bacterial surface proteins. Recently, we showed that extracellular vesicles (EVs) produced by P. freudenreichii CIRM-BIA129 mimic the immunomodulatory features of parent cells in vitro (i.e. modulating NF-κB transcription factor activity and IL-8 release) which underlies the role of EVs as mediators of the probiotic effects of the bacterium. The modulation of EV properties, and particularly of those with potential therapeutic applications such as the EVs produced by the probiotic P. freudenreichii, is one of the challenges in the field to achieve efficient yields with the desired optimal functionality. Here we evaluated whether the culture medium in which the bacteria are grown could be used as a lever to modulate the protein content and hence the properties of P. freudenreichii CIRM-BIA129 EVs. The physical, biochemical and functional properties of EVs produced from cells cultivated on laboratory Yeast Extract Lactate (YEL) medium and cow milk ultrafiltrate (UF) medium were compared. UF-derived EVs were more abundant, smaller in diameter and displayed more intense anti-inflammatory activity than YEL-derived EVs. Furthermore, the growth media modulated EV content in terms of both the identities and abundances of their protein cargos, suggesting different patterns of interaction with the host. Proteins involved in amino acid metabolism and central carbon metabolism were modulated, as were the key surface proteins mediating host-propionibacteria interactions.Importance Extracellular vesicles (EVs) are cellular membrane-derived nanosized particles that are produced by most cells in all three kingdoms of life. They play a pivotal role in cell-cell communication through their ability to transport bioactive molecules from donor to recipient cells. Bacterial EVs are important factors in host-microbe interactions. Recently we have shown that EVs produced by the probiotic P. freudenreichii exhibited immunomodulatory properties. We evaluate here the impact of environmental conditions, notably culture media, on P. freudenreichii EV production and function. We show that EVs display considerable differences in protein cargo and immunomodulation depending on the culture medium used. This work offers new perspectives for the development of probiotic EV-based molecular delivery systems, and reinforces the optimization of growth conditions as a tool to modulate the potential therapeutic applications of EVs.

The role of Propionibacterium freudenreichii in mitigating inflammation has been a subject of study for many years. Lately, it has been shown that immunomodulation properties are strain- specific and that the major responsible for inflammatory modulation ofimmunomodulation by strain CIRM-BIA129 is the presence of the surface layer protein B (SlpB), which has presented immunomodulation even when expressed in other bacteria. Extracellular vesicles (EVs) are nanosized spherical structures, produced by organisms of all kingdoms, including bacteria. They have been associated with inter-organism communication, pathogenesis, competition, and immunomodulation. Recent studies were aiming to address the role of EVs, in the probiotic effects of bacteria. The properties of P. freudenreichii CIRM-BIA129- derived EVs have been investigated. EVs produced by CIRM-BIA129 cultured in milk ultrafiltrate medium (UF) have been characterized by regarding size and morphology. UF-derived EVs displayed a monodisperse pattern with a modal size of 84.80 ± 2.34 nm. They are composed of a wide variety of proteins, mainly involved in metabolic processes, cellular processes and signaling, and storage and processing of information. A, among these proteins, SlpB was found in high abundance. P. freudenreichii EVs were able to inhibit, in a dose dependent manner, the increase of IL-8 production in HT-29 cells induced with LPS, due to NF-KB pathway inhibition, without causing cell cytotoxicity. EVs produced by a CIRM-BIA129 mutant strain with a knockout for SlpB showed a less efficient reduction in IL-8 production. Results have shown that the environmental conditions are able to modify EVs content and, consequently, their immunomodulatory effects. A change in the growth medium, from UF to YEL (yeast extract-lactate) showed a lower production of EVs, with a slightly larger size. EVs produced in YEL did not perform as well in the inhibition of the NF-KB pathway and had no effect on IL-8 production. Recent results have shown that P. freudenreichii EVs were able to protect Caco-2 cells from inflammation-induced excessive increasing permeability. These Altogether, these results show that Evs produced by beneficial propionibacteria are able to generate immunomodulationtrigger immunomodulation, similar to the parental strain. As for the SlpB role in EVs immunomodulation

Les phénotypes et les habitats microbiens ont été largement explorés au cours de ces 50 dernières années grâce à des inventaires d’espèces par des méthodes culturales ou impliquant du séquençage massif. Les phénotypes et les fonctionnalités assurés par les micro-organismes sont décrits aussi bien dans la littérature que dans des bases de données spécialisées. Ces données sont difficiles à atteindre et à croiser. Notre objectif était de (i) développer un pipeline bio-informatique (ii) de concevoir et d’implémenter une base de données - Omnicrobe- permettant un accès unifié à toutes ces données (iii) de valider biologiquement son usage.Le pipeline extrait de l’ensemble du corpus PubMed, le nom vernaculaire des taxons microbiens (bactéries, levures, champignons, microalgues, virus), des habitats, des phénotypes, des fonctionnalités et des relations liant ces différentes entités, en utilisant des outils performants de fouille de texte automatisée. Les données des collections internationales du CIRM, de la DSMZ et de Genbank ont été ajoutées au jeu de données extraites. Les différentes entités ont été catégorisées en utilisant la taxonomie NCBI et une ontologie ad hoc des habitats et des phénotypes (http://agroportal.lirmm.fr/ontologies/ONTOBIOTOPE/, [1]). L’ensemble des données a été rassemblé, structuré et implémenté dans une base de données relationnelle publique (https://omnicrobe.migale.inrae.fr/) permettant des requêtes multi-critères sur mots clés via une interface graphique ainsi que l’accès direct aux résumés des articles dont est tirée l’information. Omnicrobe a été utilisé avec succès pour concevoir rapidement un ferment pour réaliser des yaourts à base de jus de soja [2], sur des critères d’innocuité et d’acidification. Outre son usage favorisant l’innovation alimentaire, Omnicrobe facilite la compréhension du fonctionnement des écosystèmes microbiens en affichant avec exhaustivité l’ensemble des phénotypes et des fonctionnalités des espèces présentes.

The FAO recommends the digestible indispensable amino acid score (DIAAS) as the measure for protein quality, for which the true ileal digestibility needs to be assessed in humans or pigs. However, due to high costs and ethical concerns, the FAO strongly encourages as well the development of validated in vitro methods, which complement the in vivo experiments. Method: Recently, an in vitro workflow, based on the validated static INFOGEST protocol, was developed and compared towards in vivo data. In parallel to the validation with in vivo data, the repeatability and reproducibility of the in vitro protocol were tested in an international ring trial (RT) with the aim to establish an international ISO standard method within the International Dairy Federation (IDF). Five different dairy products (skim milk powder, whole milk powder, whey protein isolate, yoghurt, and cheese) were analyzed in 32 different laboratories from 18 different countries, across 4 continents. Results: in vitro protein digestibilities based on Nitrogen, free R-NH2, and total amino acids as well as DIAAS values were calculated and compared to in vivo data, where available. Conclusion: The in vitro method is suited for quantification of digestibility and will be further implemented to other food matrices.

Dans l'ensemble de la chaîne de production alimentaire, de la ferme à la fourchette, les étapes de transformation des aliments ont un impact environnemental important. Pour accompagner les nécessaires changements drastiques des systèmes alimentaires vers plus de durabilité, il est nécessaire d’apporter des données environnementales sur les produits transformés et de réfléchir à l’amélioration et l’éco-conception des procédés.L’Analyse de Cycle de Vie, ACV est une méthode standardisée par les normes ISO, de plus en plus utilisée, qui traite des aspects et des impacts environnementaux potentiels tout au long du cycle de vie d’un produit. Cette présentation abordera l’apport de l’ACV dans l’évaluation environnementale des procédés laitiers et des produits transformés qui en sont issus. Les potentialités de l’ACV (identification d’opérations à fort impact environnemental, évaluation de propositions alternatives innovantes, ...) seront illustrées au travers d’exemples du secteur laitier. La présentation abordera également les approches émergentes et prometteuses de conception environnementale des procédés incluant l’ACV et les challenges et travaux ultérieurs qui en découlent (collecte et structuration des données, modélisation des procédés, intégration des autres dimensions de la durabilité, intégration des approches dans la filière, ...).

Staphylococcus aureus is an opportunistic pathogen that causes a range of devastating diseases including chronic osteomyelitis, which partially relies on the internalization and persistence of S. aureus in osteoblasts. The identification of the mechanisms of the osteoblast response to intracellular S. aureus is thus crucial to improve the knowledge of this infectious pathology. Since the signal from specifically infected bacteria-bearing cells is diluted and the results are confounded by bystander effects of uninfected cells, we developed a novel model of long-term infection. Using a flow cytometric approach we isolated only S. aureus-bearing cells from mixed populations that allows to identify signals specific to intracellular infection. Here we present an in-depth analysis of the effect of long-term S. aureus infection on the transcriptional program of human osteoblast-like cells. After RNA-seq and KEGG and Reactome pathway enrichment analysis, the remodeled transcriptomic profile of infected cells revealed exacerbated immune and inflammatory responses, as well as metabolic dysregulations that likely influence the intracellular life of bacteria. Numerous genes encoding epigenetic regulators were downregulated. The later included genes coding for components of chromatin-repressive complexes (e.g., NuRD, BAHD1 and PRC1) and epifactors involved in DNA methylation. Sets of genes encoding proteins of cell adhesion or neurotransmission were also deregulated. Our results suggest that intracellular S. aureus infection has a long-term impact on the genome and epigenome of host cells, which may exert patho-physiological dysfunctions additionally to the defense response during the infection process. Overall, these results not only improve our conceptual understanding of biological processes involved in the long-term S. aureus infections of osteoblast-like cells, but also provide an atlas of deregulated host genes and biological pathways and identify novel markers and potential candidates for prophylactic and therapeutic approaches.

The aim of LOCALNUTLEG project is to empower local Mediterranean nuts and legumes with quality characteristics that depend exclusively on the territory on which they are produced through the development of nniovative plant-based food products, fermented or not. To do so, LOCALNUTLEG will promote local Mediterranean nuts and legumes with a legal registered trademark (Protected Designation of Origin (PDO) or Protected Geographical Indication (PGI)) or autochthonous identity and will represent a potential opportunity to Mediterranean countries to balance the sustainability of Agri-food resources and dietary habits considering the economical and health impact of the new developed products. The project aims to develop innovative food products by the identification of local nut and legumes varieties tied to Mediterranean gastronomic culture, providing the maximum amount of nutrients and bioactive compounds, and to promote Mediterranean diet through plant-based food products., Le but de LOCALNUTLEG est d’autonomiser des filières locales méditerranéennes de produits végétaux, fermentés ou non, sous signe de qualité géographique ou avec une identité locale forte. Pour cela le projet développera des produits alimentaires innovants à partir des variétés locales de fruits secs et légumes secs liés à la culture gastronomique méditerranéennes, riches en nutriments et composés bioactifs. Ces produits seront une opportunité de conjuguer la durabilité des matières premières agricoles locales et des habitudes alimentaires pour des produits sains et contribuant à l’économie locale.

La microfiltration tangentielle de lait écrémé, MF 0.1 μm, équipée de membranes céramiques, est apparue à l’échelle industrielle au milieu des années 90s pour la séparation des micelles de caséines (CN) et des protéines sériques (PS). Face aux enjeux grandissants des ingrédients laitiers et l’importance que revêt la MF 0.1 μm dans la production de ces ingrédients, notamment en nutrition infantile, il devient important d’optimiser la MF 0.1 μm de lait écrémé, que ce soit au niveau de la conception de l’installation que de sa conduite. Le développement industriel a été rendu possible grâce à la mise en œuvre du système Uniform Transmembrane Pressure (UTP) qui permet de limiter l’accumulation de la matière à la membrane et donc de maintenir de très bonnes performances en termes de flux de perméation et de transmission de protéines sériques au cours du temps de production [1]. Néanmoins, les coûts énergétiques et économiques (investissement et maintenance) de cette opération sont très élevés puisque la mise en circulation du perméat à co-courant du rétentat nécessite une pompe supplémentaire côté perméat. Au début des années 2000, le concept à Gradient de Perméabilité (GP) a été proposé par Pall pour rendre la MF 0.1 μm plus économiquement attractive. Ce concept offre des propriétés de perméabilité (et de résistance hydraulique) du support macroporeux variables entre l’entrée et la sortie de la membrane, de manière à compenser les gradients de pression transmembranaire et ainsi offrir des performances acceptables sans ajout de pompe côté perméat et avec une couche filtrante de 0.1μm identique à la membrane UTP. A l’heure actuelle, les membranes UTP et GP cohabitent industriellement pour la mise en œuvre de la MF 0.1 μm. Pour des raisons principalement économiques, la membrane GP remplace peu à peu la membrane UTP. Elle est proposée depuis une quinzaine d’années par les équipementiers qui se basent principalement sur leur expérience, savoir-faire et connaissances empiriques pour proposer le dimensionnement et les modes de conduites de ces installations. Pourtant peu d’études sont publiées sur les performances de la membrane GP (une dizaine d’articles seulement dont [2]) et l’analyse de ces articles montre le manque de connaissances sur les performances de cette membrane. La conception des installations est donc réalisée sans une réelle analyse comparative critique. L’objectif de ce travail est d’apporter des connaissances sur les performances de la membrane GP 0.1μm pour la séparation CN-PS de lait écrémé thermisé de vache à 50°C et de comparer ces performances à celles de la membrane UTP, qui fait référence. Les performances des membranes Pall 0.1 μm UTP et GP ont été étudiées à 3 niveaux de Facteur de Réduction Volumique (FRV1 ; 2 ; 3). La membrane GP a été mise en œuvre dans les conditions d’écoulement tangentiel pour lesquelles elle est conçue, c’est à dire une Perte de Charge Rétentat (PCR) de 2 bar, conduisant à des vitesses d’écoulement tangentiel de l’ordre de 7.0 m/s. Pour chaque facteur de réduction volumique (FRV), les membranes GP et UTP ont été comparées à la même perte de charge (2 bar). Pour la membrane GP, les plages de pression transmembranaire (PTM) préconisées sont supérieures à 1.0 bar pour éviter la rétroperméation. Pour la membrane UTP, les plages de PTM préconisées sont inférieures à 0.6 bar pour limiter le colmatage. D’un point de vue hydrodynamique, le flux de perméation Jp (l/h/m²) a été suivi en fonction de la PTM appliquée par palier de 15 à 30 min à des valeurs comprises entre 0.4 et 0.8 bar pour la membrane UTP et entre 0.8 et 2.0 bar pour la membrane GP. D’un point de vue sélectivité, la transmission des PS (TrPS) a été calculée à partir des concentrations en protéines sériques des fractions rétentat (CrPS) et perméat (CpPS) déterminées par la méthode Kjeldahl. TrPS (%) = (CpPS / CrPS) x100. Les fuites de micelles de caséines dans les perméats ont été évaluées par mesure de colmatage. ✓ Pour chaque FRV, les valeurs de Jp limite sont similaires pour les 2 membranes : 180 l/h/m² à FRV1 ; 110 l/h/m² à FRV2 (Fig 1a). A FRV1, Jp limite est atteint à une PTM respectivement de 2.0 bar pour la membrane GP et 0.4 bar pour la membrane UTP. ✓ Pour la membrane UTP, TrPS (%) diminue de 80 à 68 % sur la plage de PTM de 0.4 à 0.8 bar, et cela indifféremment pour FRV 1 et 2. En revanche, pour la membrane GP, TrPS est stable et élevée (80%) de 0.8 à 1.8 bar à FRV 1 mais elle est faible et diminue de 65% à 55% sur la même plage de PTM, à FRV 2 (Fig 1b). ✓ A la différence des perméats obtenus avec la membrane UTP qui sont limpides (turbidité = 0 NTU), les perméats obtenus avec la membrane GP sont troubles (turbidité ≈ 100 NTU) et indiquent des fuites de micelles de caséines estimées à 0.2 g/kg de caséine. Fig 1 - Performances des membranes GP (triangle) et UTP (rond) en fonction de la pression transmembranaire (PTM) à FRV1 (en vert) et FRV2 (en orange) pour une PCR 2 bar, à 50°C Fig1a : Flux de perméation Jp ; Fig1b : Transmission de protéines sériques, TrPS Ces résultats montrent que le choix de la membrane GP (et de son mode de conduite à PCR = 2bar) a un impact sur les performances de la séparation CN-PS. A FRV élevé comme recherché industriellement (FRV 2), les flux sont plus élevés avec la membrane UTP dans les conditions de PTM préconisées (1.0 +/-0.2 bar pour GP et 0.6 +/-0.2 bar pour UTP). Les performances en termes de transmission de PS sont semblables (68±5 %) à FRV 2, mais la fuite de micelles est plus élevée, ce qui pénalise la valorisation du perméat. Ces différences s’expliquent par la conception même de la membrane GP. Ces connaissances permettent d’apporter des informations indispensables pour envisager la conception raisonnée et optimisée des installations de MF mettant en œuvre des membranes céramiques [3]

Les quatre missions canoniques d’un CRB sont la collecte, la caractérisation, la conservation et la distribution des ressources biologiques. Pour chacun de ses items, qui sont les chapitres de ce n° spécial avec les fonctions support, des innovations sont mises au point par les agents des CRB. L’Infrastructure de Recherche RARe (Ressources Agronomiques pour la Recherche) structure les Centres de Ressources Biologiques de son périmètre dans 5 Piliers : animal, végétal, microbien, environnement et forêt. Elle prouve par ce numéro spécial, issu de sa volonté de synergie entre ses Piliers et ses CBR, qu’elle peut s’appuyer sur les compétences diversifiées de ses collaborateurs, leur créativité et leur ingénierie.; National audience; Les missions des Centres de ressources biologiques (CRB), quels que soient leurs objets d’intérêt, sont de collecter, authentifier, conserver et distribuer des ressources biologiques. Ici nous nous intéressons à la mission de collecte, à travers l’exemple du CIRM (Centre international de ressources microbiennes), dont les catalogues de micro-organismes rassemblent des collections spécialisées, si précieuses pour la recherche. Mais pourquoi et comment enrichir les collections ? Nous apportons quelques réponses à cette question, à l’appui d’exemples tirés de récentes expériences des CRB du CIRM.

This poster serves to underline the following: • we use synchrotron radiation to measure local solids concentration in the membrane fouling layer (concentration vs. distance to the membrane with the spatial resolution of 20 µm); this measurement requires less than a minute; • our method of fouling analysis is particular, but our conclusions are general; • fouling is sensitive to all operating conditionsnot only that that applied during the fouling formation, but also that applied during the fouling analysis; • the fouling layer may change during the fouling analysis; this change is not immediate; this fact must be accounted in the fouling analysis (whatever is the method of analysis); • all experiment conditions should be described properly at every stage of experiment (e.g. no such thing as "simple rinsing")

L’augmentation des sciences et recherches participatives (SRP) depuis les années 2000 est inédite, même si la participation de « non-scientifiques » à la production de connaissances scientifiques n’est pas nouvelle. Cette présentation fera une brève introduction sur les aspects historiques et l’état des lieux sur les SRP en France. Les SRP impliquent des acteurs de la société civile, dont le type et les modalités de participation ainsi que le degré d’implication sont très variables. L'implication de ces acteurs peut aller du crowdsourcing, c’est-à-dire la fourniture d’informations et éventuellement d’échantillons, jusqu’à la participation aux expérimentations, par exemple la formalisation de la question de recherche, la co-construction du design expérimental, la co-réalisation des expérimentations, le partage des couts. Trois projets de SRP portant sur des aliments fermentés seront détaillés : l’ANR Bakery sur le pain au levain, le projet européen sur le Gwell, un lait fermenté traditionnel breton, et le projet régional FLEGME sur les légumes fermentés. A travers ces trois exemples, nous comparerons le contexte, les questions de recherche de ces projets, les dispositifs de recherche mis en place pour y répondre et les modalités d’interaction avec les collectifs. Quelques résultats marquants seront présentés pour chaque projet. Une place privilégiée sera faite aux retours d’expérience des porteurs de ces projets ainsi que d'autres projets de SRP conduits à INRAE, pour en souligner les spécificités et éclairer à la fois les points de vigilance, les difficultés, l'intérêt et les bénéfices de cette approche.

SYALSA : Systèmes alimentaires et santé humaineAu cœur des relations entre agriculture, alimentation et environnement, les questions de santé peuvent être un moteur pour transformer les systèmes alimentaires. L’objectif général de SYALSA est d’identifier et d’évaluer les leviers d’action susceptibles de rendre les systèmes alimentaires plus favorables à la santé humaine, à travers l’alimentation, mais aussi à travers leurs effets sur l’environnement, en prenant en compte les co-bénéfices entre santé et environnement. Le métaprogramme vise ainsi à mieux comprendre les divers facteurs et mécanismes d’interaction qui, depuis la production agricole jusqu’à la consommation alimentaire, affectent la santé humaine. Il explore les relations entre les pratiques de production et de transformation, les expositions des populations à des contaminants environnementaux, et les impacts sur la santé. Enfin, il s’attache à caractériser, évaluer et accompagner les changements manifestes ou potentiels (innovations biotechniques, politiques publiques, comportements d’acteurs) susceptibles d’améliorer les bénéfices des systèmes alimentaires sur la santé des individus et des populations. Lancé en janvier 2021, SYALSA mobilise les réflexions de la prospective scientifique interdisciplinaire « Nexus Santé »

Le CBL (Club des Bactéries Lactiques) est une manifestation scientifique qui réunit chercheurs, enseignants-chercheurs et industriels R&D, pour échanger sur les avancées scientifiques et techniques réalisées dans le domaine des bactéries lactiques. Les thèmes abordés sont le reflet des larges potentiels de ces bactéries et de leurs applications : fermentation alimentaire, santé humaine et animale, probiotique, environnement…; International audience; Breastfeeding is recommended by the WHO for the first 6 months of life. It provides health benefits to infants compared to infant formula (IF), including protection against intestinal and respiratory infections in early childhood and a lower risk of metabolic and immune diseases later in life. These differences in health benefits are probably due to the difference in composition between IF and human milk (HM), the latter being more complex and richer in bioactive components. Our hypothesis is that HM health benefits come in part from the HM microbiota -a paucimicrobial ecosystem that presents a great bacterial diversity- through a role on gut immune homeostasis.This study aimed to better understand the immunomodulatory role of HM bacteria. To this end, bacteria were isolated from healthy breast milks and screened for their immunomodulatory potential on two different cellular models in order to select candidates with probiotic interest.The collection of 28 breast milks at the Rennes Sud University Hospital allowed the constitution of a strain library of 1164 isolates with 29 genera and 56 different species. From this library, 88 isolates selected for their prevalence in HM microbiota were first screened on PBMC (blood mononuclear cells) for the production of IL-10 and TNF-alpha cytokines. Among them, 28 isolates exhibiting interesting immunomodulatory properties on PBMC model underwent a second screening on a quadricellular model of intestinal epithelium including enterocytes (Caco2), caliciform cells (HT 29-MTX), M cells (differentiated Caco2) and macrophages (THP-1). This study showed the species- and strain-dependence of HM bacteria immunomodulatory potential and revealed interesting properties of HM bacteria in the context of infant immune system maturation. This study is part of the PROLIFIC project funded by the Régions Bretagne and Pays de la Loire and by the BBA MilkValley industrial consortium.

Objectives : Buriti (Mauritia flexuosa) is an under-utilized Amazonian fruit with a high content in phenolic compounds. Although polyphenols have health benefits, they may also hinder protein digestibility but salivary proteins might counteract this negative effect through binding to polyphenols. The objective of this work was to describe the impact of saliva on protein digestibility of yoghurt containing buriti.Methods : A water extract of buriti flour was mixed with human saliva (LeeBiosolutions) at estimated concentrations varying from 0.001 to 0.05 g polyphenols (GAE)/ml. The particle size distribution in the mixtures was measured by laser diffraction. Pepsin and trypsin activity were measured in presence or not of the buriti extract. Furthermore, an in vitro model of digestion was applied to yoghurt or to yoghurt with buriti flour (1:4 w/w). In the oral phase of digestion, the two food products was mixed with human saliva or simulated salivary fluid SSF (ionic solution) at a ratio of 1:1. The standardized INFOGEST static model was then performed. The gastric and intestinal phases of digestion were conducted using pepsin and pancreatin, respectively. Digestive proteolysis was assessed by measuring accessible NH2 groups in samples over time by the OPA method.Results : Buriti extract also had no significant effect on pepsin or trypsin activity. However, when added to yoghurt, buriti flour had a large inhibiting impact on digestive intestinal proteolysis. Interestingly, the presence of saliva instead of SSF during digestion of buriti-containing yoghurt increased slightly the digestive proteolysis, both in the gastric and intestinal phase. Finally, buriti extract had no detectable impact on particle size in saliva (D50 ~ 45µm), suggesting no major aggregation of salivary proteins.Conclusions : Compared to SSF, saliva improved protein digestibility of yoghurt containing buriti flour, but is unclear whether this occurred through binding of salivary proteins to polyphenols. Further investigations are required.

Les interactions impliquant les bactéries lactiques (BL), responsables des propriétés sanitaires, organoleptiques et santé des produits fermentés, sont souvent étudiées en association avec des levures ou des bactéries propioniques dans divers produits alimentaires fermentés et les mécanismes de leurs interactions sont assez bien caractérisés. En ce qui concerne les interactions entre BL, elles ont principalement été étudiées pour tester des interactions antagonistes. Comprendre comment les BL peuvent interagir positivement serait utile dans de multiples domaines liés à l'alimentation, comme la production de produits fermentés aux propriétés fonctionnelles améliorées ou la fermentation de nouvelles matrices alimentaires. Les dépendances nutritionnelles, notamment celles vis-à-vis des sources d'azote, régissent de nombreuses interactions positives microbiennes. De telles interactions positives ont déjà été étudiées entre les BL du yaourt. Cependant, elles n'ont jamais été exploitées pour créer des co-cultures de BL issues de biotopes différents. L'objectif de cette étude était de favoriser les interactions positives entre BL, basées sur les dépendances aux sources d’azote dans les co-cultures pour augmenter les taux d'acidification, la consommation de sucres comme le raffinose et la production de composés volatils. La stratégie consistait à exploiter les activités protéolytiques et les auxotrophies des acides aminés des BL. Un milieu chimiquement définia été développé pour permettre spécifiquement la croissance des six souches de BL utilisées, trois protéolytiques et trois non protéolytiques. Chacune des souches protéolytiques, Enterococcus faecalis CIRM-BIA2412, Lactococcus lactis NCDO2125 et CIRM-BIA244, a été cultivée avec chacune des souches de BL non protéolytiques : L. lactis NCDO2111, Lactiplantibacillus plantarum CIRM-BIA465 et CIRM-BIA1524. La croissancebactérienne a été suivie à l'aide de chambres compartimentées pour comparer la croissance en mono- et co-cultures. Chaque souche protéolytique a induit différents types d'interactions 2 : fortement positives avec E. faecalis CIRM-BIA2412, faiblement positives avec L. lactis NCDO2125, ou nulles avec L. lactis CIRM-BIA244. En co-cultures directes, les interactions ont conduit à des taux d'acidification plus rapides, à un pH final plus bas, à une utilisationplus élevée du raffinose et à des concentrations plus élevées en cinq composés volatils, pouvant donner des arômes intéressants. Les fortes interactions étaient associées à des concentrations plus élevées en tryptophane, valine, phénylalanine, leucine, isoleucine et en peptides. Une analyse peptidomique a permis d’identifier les acides aminés hydrophobes et à chaîne ramifiée comme essentiels dans ces interactions. Cette étude donne un nouvel aperçu des mécanismes régissant les interactions entre BL et offre des solutions pour améliorer ou concevoir des ferments lactiques.

Les microalgues sont des organismes unicellulaires photosynthétiques riches en nutriments constituant une biomasse à fort potentiel pour des applications alimentaires. Leur production est énergivore avec un fort risque d'altération au cours du procédé de récolte et de conservation. La fermentation, procédé ancestral de conservation, offre une alternative aux procédés actuels en limitant le développement des flores d'altération. La capacité des Bactéries Lactiques (BL) à fermenter les microalgues n'a été jusqu'à présent que très peu explorée1,2.Dans cette étude, nous avons criblé la capacité de 89 souches (10 espèces) de BL à fermenter Chlorella vulgaris, une des deux microalgues autorisées en alimentation humaine3. Pour cela nous avons suivi au cours du temps une suspension de biomasse lyophilisée de C. vulgaris resolubilisée (50mg/mL) inoculée par les BL en mesurant le pH et en dénombrant les flores bactériennes présentes. En parallèle, nous avons évalué la capacité fermentaire des 89 souches à métaboliser huit sucres. Pour les trois souches présentant la meilleure aptitude à fermenter C. vulgaris, nous avons testé l'effet de l'ajout de différentes concentrations de glucose et d'extrait de levure sur la baisse de pH.Les résultats obtenus montrent que 72% des souches acidifient la biomasse microalgale selon 3 profils d’acidification : A - acidification rapide en moins de 12h (60 % des souches) ; B - vitesse d'acidification intermédiaire nécessitant 36h de fermentation pour avoir une baisse significative du pH (12%) ; C - absence d'acidification au bout de 36h. Les souches sont réparties indépendamment de l'espèce dans les trois profils. Les espèces les plus acidifiantes sont L. lactis, L. plantarum, L. pentosus avec un pH moyen de 4.86 à 36h. Aucun lien n’a pu être observé entre la capacité fermentaire des 89 souches pour les huit sucres testés et leur capacité à fermenter les microalgues. Pour les trois souches les plus acidifiantes (CIRM-BIA2469, CIRM-BIA2211 et CIRM-BIA2178), seul l’ajout de 1g/L de glucose a eu pour effet d'améliorer l'acidification. Une flore contaminante (identifiée par séquençage ADNr16S comme Bacillus sp.), présente à 3.103 UFC/mL dans les lots de C. vulgaris testés croit jusqu'à 108 UFC/mL en l’absence de BL. Elle est en revanche systématiquement inhibée lors de la fermentation par les BL.Nos travaux montrent que les BL sont capables de fermenter la microalgue C. vulgaris et d'inhiber le développement de la flore contaminante. C'est à notre connaissance la première étude de fermentation de microalgues portant sur une aussi grande diversité de BL.

Complex coacervation is a liquid-liquid phase separation that occurs between two oppositely charged macromolecules and leads to the formation of a concentrated phase called coacervates. The formed coacervates are proposed as efficient encapsulating agents for bioactives. Here, we studied the specific case of heteroprotein complex coacervation (HPCC) between positively charged Lactoferrin (LF) and negatively charged β-lactoglobulin (β-LG). We aimed to determine the sensitivity of LF/β-LG complex coacervation process to ionic strength and subsequent properties of formed coacervates. Dialysis experiments showed the very high sensitivity of LF/β-LG complex coacervation to ionic strength. Complex coacervation only occurred at ionic strength values lower than 30 mM. Furthermore, in an environment of low ionic strengths, there is a small window wherecomplex coacervation between the two proteins is favoured as assessed by ITC measurements. The interaction strengths deduced from the ITC profiles in the range of 3-5 mM NaCl are higher than in the absence of salt. Hence, addiction of a low salt concentration promoted the interaction between the two proteins leading to a higher coacervation yield. These results shed new light on the mechanisms governing the complex coacervation in heteroprotein systems

In a general context of food transition, plant-based fermented products experience a considerable development. This results from evolution of consumers’ habits, demanding healthy products with a low environmental impact. This requires research and development, and thus higher scientific knowledge. Fermented soy milks constitute a favorable alternative source of live probiotics. Lactobacillus delbrueckii subsp. bulgaricus and Propionibacterium freudenreichii are extensively used in probiotic dairy products. Their adaptation to the dairy environment has been extensively studied and documented. However, little is known about their adaptation to the soy environment. We investigated suitability of soymilk as a substratefor growth and acidification by Lactobacillus delbrueckii CIRM-BIA1592 and Propionibacterium freudenreichii CIRM-BIA129. A specific focus was made on changes in term of proteome composition, cellular morphology in comparison with the cultivation in cow milk. In order to understand such adaptation, we explored their growth and survival in soy milk ultrafiltrate and compared it to cow’s milk ultrafiltrate. The two species grew in cow milk,yet not in soy milk. P. freudenreichii did grow in soy milk when co-cultured with the lactic acid bacterium L. plantarum while L. delbrueckii did not, even in co-culture with S. thermophilus. Our results revealed major differences in bacterial morphology and proteome composition. Lactobacilli upregulated stress proteins, and downregulated cell cycle and division machinery. Regarding propionibacteria, differential expression of proteins involved in amino acid metabolism, in carbohydrate metabolism, and in stress remediation was observed. These results suggest that changing the fermented substrate may significantly affect both technological and probiotic properties of conventional dairy starters. This should be considered for the development of new fermented functional foods. Screening of a large collection of lactic and propionic acid bacteria is in progress, based on their technological and probiotic properties. Bacterial stains and consortia thereof will be selected for the production of new plant-based fermented functional foods