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[hal-04213765] Beef protein ingredients from fat rendering process are promising functional ingredients
The valorization of co-products may be a promising way to meet the dual challenge of increasing global food resources and sustainability of food systems. In particular, meat co-products may be nutritionally interesting protein resources, if they offer functional properties in accordance with food applications. In that aim, two bovine co-products, resulting from the fat rendering process, have been characterized, regarding the protein solubility, gelling, and emulsifying properties. The effect of protein concentration, pH variation and NaCl addition on these properties was tested. Despite an effect of the ionic strength on the protein solubility of the two ingredients, a little or no significant impact was observed on the functionalities. Similarly, the functional properties were scarcely affected by pH. In the end, the protein concentration has proven to be the only important parameter, which points to an easy utilization of these ingredients in many food conditions.
ano.nymous@ccsd.cnrs.fr.invalid (Rozenn Le Foll) 21 Sep 2023
https://hal.inrae.fr/hal-04213765
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[hal-04317337] Human milk bacteria individually or as a synthetic community exhibited contrasted immunomodulatory profiles and impact on the gut epithelial barrier
Breastfeeding is recommended for the first 6 months of life. Many bioactive compounds of the human milk (HM) support the development of the intestinal immune system and barrier functions in infants. Our hypothesis was that HM microbiota contributes to these health benefits. Our objective was to characterise in vitro the role of HM bacteria, either individually or combined in synthetic communities (SynCom), on gut homeostasis. A collection of bacterial isolates, reflecting HM microbiota composition, was made from 28 healthy mothers exclusively breastfeeding. Firstly, the immunomodulatory profile of 84 HM bacterial isolates belonging to 38 species was characterized using blood mononuclear cells (PBMC). Secondly, the impact of a subset of 29 strains was deeply investigated on epithelial immune and barrier functions using a quadricellular (Caco2, HT29-MTX-E12, M cell, THP1 cells) model of the intestinal epithelium. Strains were characterized for their ability to modulate cellular IL-10 and TNF-α production and the expression of genes related to the barrier, immune and apoptosis/proliferation functions. Based on these results, 2 SynComs were designed and characterized on the quadricellular model. HM bacteria displayed a large range of immunomodulatory properties. Using MultiDimensionate Scaling (MDS) on IL-10 and TNF-α production by PBMC, isolates were classified into 5 groups with specific signatures, highlighting the anti- and/or pro-inflammatory profiles of HM bacteria. Further, the MDS analysis of cytokine production and gene expressions of the quadricellular model stimulated by each of the 29 bacteria, classified strains into 3 groups named Quadri1, 2 and 3, according to their immunomodulatory activity and their impact on the epithelial barrier function. The composition of each group did not reveal major taxonomic biases between the 3 groups, but a diversity of the HM bacteria impact on gut epithelium within each genus or species. Quadri3 and, to a lesser extent, Quadri1 strains stimulated the immune function whereas Quadri2 hardly affected it. Besides, Quadri1 and 3 strains reinforced the epithelial barrier whereas an opposite effect was observed with Quadri2 strains. Finally, strains belonging to prevalent HM genera and with contrasted immunomodulatory profiles were assembled in two HM-like SynCom of 11 strains. The 2 Syncoms displayed different immunomodulatory properties, yet less contrasted than individual strains, whereas they both exhibited beneficial impact on barrier function. This study showed the great diversity of immunomodulatory potential and impact on the barrier function of HM bacteria, individually or assembled in SynCom, highlighting the potential of the HM microbiota to modulate the intestinal development.
ano.nymous@ccsd.cnrs.fr.invalid (Charles Le Bras) 01 Dec 2023
https://hal.inrae.fr/hal-04317337
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[hal-04317931] Human milk metabolites are bioactive and can modulate gut physiology : in vitro study in a pluricellular model of intestinal epithelium
Human milk (HM) is associated with major short- and long-term health benefits for infants. However, infant formulas (IF), substitutes for HM, are widely used for infant nutrition. Although these IFs meet the nutritional needs of newborns, they are devoid of many bioactive compounds present in HM, such as immunoglobulins, hormones and a multitude of metabolites. HM metabolites include short chain fatty acids (SCFAs) (butyric acid, acetic acid, propionic acid), polyamines (putrescine, spermine, spermidine), tryptophan derivatives (indole, indole-lactic acid, kynurenine) as well as GABA and lactate. These metabolites are known to be produced within the gastrointestinal tract by the intestinal microbiota and to have effects on the host physiology but their relevance in HM has not been studied so far. The objective of our study was therefore to decipher the effects of HM metabolites on intestinal physiology using an in vitro pluricellular and polarized model of intestinal epithelium, including Caco-2 (as enterocytes), HT-29 MTX (as goblet cells), NCI-H716 (as enteroendocrine cells) and M cells. HM metabolites were studied at concentrations close to those found in HM, that is quite low. Their effects on trans-epithelial electric resistance (TEER) and on the expression of genes involved in the intestinal barrier, immune, antioxidant, endocrine and digestive functions were analyzed. SCFAs strongly modulated different intestinal functions, particularly the immune one with a significant downregulation of genes coding for IL-8, MyD88 and TFF3. They also modulated genes encoding tight junctions, as did GABA and polyamines, upregulating CLDN3, TJP1 and CLDN4 respectively and downregulating CLDN1 for SCFA and polyamines and CLDN7 for GABA. In parallel SCFAs significantly increased TEER, highlighting a potential reinforcing effect on the epithelial barrier while polyamines and GABA had no effect on TEER. Finally, SCFAs, GABA and lactic acid modulated the expression of some transporters involved in nutrition such as MCT1, GLUT1 and SGLT1 respectively. HM metabolites, despite their low concentration, are able to impact the intestinal barrier physiology, inviting us to consider their implementation in IF for a better mimicry of HM health benefits.
ano.nymous@ccsd.cnrs.fr.invalid (Sarah Blanchet) 01 Dec 2023
https://hal.inrae.fr/hal-04317931
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[hal-04302538] Fermented dairy and legume-based food products: from smart design of lactic acid bacteria to innovative products
For reasons linked to lifestyle, health, nutrition and sustainability, our diets are tending to change, in particular by rebalancing animal and plant sources in Western diet. This leads to the development of new food products, particularly fermented ones. Actually, fermentation can confer hedonic, hygienic and probiotic properties to fermented products. Fermented dairy products are well known, while legume-based fermented dairy analogues are less familiar. The development of legume-based fermented products requires a combination of knowledge of the raw material and of bacterial potential, in order to select bacterial strains with appropriate functions. In our study, new fermented products were developed based on milk, on plant-based analogues, and on the mixes thereof, fermented by lactic acid bacteria and/or propionic acid bacteria, selected on their capability to hydrolyze carbohydrates, proteins and on their immunomodulatory properties. By taking advantage of the functional complementarity between strains, we designed bacterial consortia capable of transforming different raw materials into functional fermented food products, with improved technofunctional and health properties. The development of such fermented products also raises the question of how positive interactions between bacteria can be promoted in order to improve the final qualities of these products.
ano.nymous@ccsd.cnrs.fr.invalid (Valérie Gagnaire) 23 Nov 2023
https://hal.inrae.fr/hal-04302538
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[hal-04302599] Extracellular Vesicles and Surface Layer Proteins as the post-biotic active ingredient of the probiotic bacterium Propionibacterium freudenreichii against colitis and mucositis
Gut inflammation constitutes a growing health concern in developed countries. It coincides with dysbiosis, including a lack of anti-inflammatory bacteria. As an example, propionibacteria are lacking in the microbiota of newborns which develop necrotizing enterocolitis. We investigated immunomodulatory properties of Propionibacterium freudenreichii. A screening led to the selection of P. freudenreichii CIRM-BIA129, a strain inducing high levels of regulatory IL-10 in human PBMCs. Consumption of this strain protected mice from colitis induced either by TNBS or by DSS. It alleviated severity of symptoms, modulated local and systemic inflammation, as well as colonic oxidative stress and epithelial cell damages. It further mitigated severity of mucositis induced by 5-fluorouracyl, preventing weight loss, reducing inflammation and mucosal damages. Mutation of the slpB gene, encoding a key surface layer protein, suppressed this immunomodulatory effect and the resulting slpB mutant induced a rather proinflammatory response and failed to prevent mucositis. 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. 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 (HIEC). Proteomic analysis revealed presence of surface layer (S-layer) proteins, including SlpB, in these EVs. Guanidine treatment of P. freudenreichii intact cells leads to extraction of surface proteins, which constitute the S-layer, the outmost structure of bacteria. These extracted proteins also mimic the effect of propionibacteria, inducing IL-10 in immune cells and modulating NF-κB and IL-8 in HIEC. This work emphasizes the importance of extractable surface proteins, including SlpB, and of EVs, in P. freudenreichii probiotic effects. It opens perspectives for the development of probiotic and postbiotic products aimed at decreasing inflammation.
ano.nymous@ccsd.cnrs.fr.invalid (Gwénaël Jan) 23 Nov 2023
https://hal.inrae.fr/hal-04302599
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[hal-04302652] Dairy starters as "2-in-1" probiotic microorganisms fermenting foods and modulating gut mucosal immunity
The gut microbiota plays a crucial role in the regulation of mucosal immunity and of the function of the intestinal barrier. Dysbiosis is accordingly associated with rupture of mucosal immune homeostasis, leading to inflammatory intestinal diseases. In this context, probiotic bacteria, including a new generation of intestinal probiotics, can maintain intestinal homeostasis and promote health. Surprisingly, little is known about the impact of fermented dairy products in this context, while they represent our main source of live and active bacteria. Indeed, they provide, through our daily diet, a high number of bacteria whose effect on mucosal immunity deserves attention. Among bacteria ingested in fermented dairy products, Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii are on top, as they are ingested in high concentrations (close to 109 ufc per gram of product) in fermented milks or cheeses.
ano.nymous@ccsd.cnrs.fr.invalid (Nassima Illikoud) 23 Nov 2023
https://hal.inrae.fr/hal-04302652
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[hal-04308495] Comprehensive probiogenomics analysis of the commensal Escherichia coli CEC15 as a potential probiotic strain
Background Probiotics have gained attention for their potential maintaining gut and immune homeostasis. They have been found to confer protection against pathogen colonization, possess immunomodulatory effects, enhance gut barrier functionality, and mitigate inflammation. However, a thorough understanding of the unique mechanisms of effects triggered by individual strains is necessary to optimize their therapeutic efficacy. Probiogenomics, involving high-throughput techniques, can help identify uncharacterized strains and aid in the rational selection of new probiotics. This study evaluates the potential of the Escherichia coli CEC15 strain as a probiotic through in silico, in vitro, and in vivo analyses, comparing it to the well-known probiotic reference E. coli Nissle 1917. Genomic analysis was conducted to identify traits with potential beneficial activity and to assess the safety of each strain (genomic islands, bacteriocin production, antibiotic resistance, production of proteins involved in host homeostasis, and proteins with adhesive properties). In vitro studies assessed survival in gastrointestinal simulated conditions and adhesion to cultured human intestinal cells. Safety was evaluated in BALB/c mice, monitoring the impact of E. coli consumption on clinical signs, intestinal architecture, intestinal permeability, and fecal microbiota. Additionally, the protective effects of both strains were assessed in a murine model of 5-FU-induced mucositis. Results CEC15 mitigates inflammation, reinforces intestinal barrier, and modulates intestinal microbiota. In silico analysis revealed fewer pathogenicity-related traits in CEC15, when compared to Nissle 1917, with fewer toxin-associated genes and no gene suggesting the production of colibactin (a genotoxic agent). Most predicted antibiotic-resistance genes were neither associated with actual resistance, nor with transposable elements. The genome of CEC15 strain encodes proteins related to stress tolerance and to adhesion, in line with its better survival during digestion and higher adhesion to intestinal cells, when compared to Nissle 1917. Moreover, CEC15 exhibited beneficial effects on mice and their intestinal microbiota, both in healthy animals and against 5FU-induced intestinal mucositis.
ano.nymous@ccsd.cnrs.fr.invalid (Tales Fernando da Silva) 27 Nov 2023
https://hal.inrae.fr/hal-04308495
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[hal-04317906] Post-milking application of a Lacticaseibacillus paracasei strain on the teat skin as a new microbial strategy to prevent bovine mastitis.
Bovine mastitis is an inflammation of the mammary gland generally due to an intramammary infection. Prophylactic treatments mostly rely on post-milking teat disinfection with chemical products such as iodine. However, the latter are questioned due to a possible irritation of the teat skin but also dissemination of residues in milk and environment. Besides, the antibiotics commonly used to treat mastitis are unfortunately not entirely effective and may contribute to the risk of dissemination of antimicrobial resistance, inviting us to explore alternative solutions. Here, a microbial strategy to prevent mastitis was explored, aiming to improve the barrier effect of the teat microbiota, through the application of a lactic acid bacteria (LAB) strain. Firstly, the impact of Lacticaseibacillus paracasei CIRM BIA 1542 application on the teat skin was explored on 23 Holstein cows in mid-lactation. Treatment (LAB, iodine or no treatment) was applied twice a day post-milking on the 4 quarters of healthy animals for 15 days. Blood, milk samples and teat skin swabs were collected at D1, D8, D15 and D26 to evaluate the LAB treatment impact at the microbial, immune and physiological levels. Data were analyzed using an analysis of variance based on mixed models. L. paracasei CIRM BIA 1542 was transiently present on teat skin and in foremilk during the 15 days of treatment, but not in the cisternal milk. Total microbial population on teat skin, in foremilk and cisternal milk was significantly increased in LAB-treated cows compared with iodine-treated cows (p < 0.05). However, no pathogen was found in cisternal milk. Ongoing analyses will evaluate the impact on teat skin and in foremilk microbiota using metataxonomic. In addition, LAB treatment did not trigger any major inflammatory response in the mammary gland: no significant impact was observed on milk somatic cell scores, although interleukin 8 released in milk tended to be slightly higher in LAB-treated cows compared with others. Finally, LAB treatment had no impact on the functionality and the integrity of the mammary epithelium, as revealed by no significant effect on milk yield and composition, nor on the mammary epithelial cell exfoliation rate into milk or the milk Na+:K+ ratio. Altogether, these results indicate that a topical treatment with L. paracasei CIRM BIA 1542 is safe with regard to mammary gland physiology and immune system, while impacting its microbiota, inviting us to further explore its effectiveness for mastitis prevention.
ano.nymous@ccsd.cnrs.fr.invalid (Coralie Goetz) 01 Dec 2023
https://hal.inrae.fr/hal-04317906
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[hal-04315650] Minimal processed infant formula vs conventional shows comparable protein quality and increased postprandial plasma amino acid kinetics in rats
Abstract During industrial processing, heat treatments applied to infant formulas may affect protein digestion. Recently, innovative processing routes have been developed to produce minimally heat-processed infant formula. Our objective was to compare the in vivo protein digestion kinetics and protein quality of a minimally processed (T-) and a heat-treated (T+++) infant formula. Sixty-eight male Wistar rats (21 days) were fed with either a diet containing 40% T- (n=30) or T+++ (n=30), or a milk protein control diet (n=8) during 2 weeks. T- and T+++ rats were then sequentially euthanized 0, 1, 2, 3, or 6 h (n=6/time point) after ingestion of a meal containing their experimental diet. Control rats were euthanized 6 h after ingestion of a protein-free meal to determine nitrogen and amino acid endogenous losses. Nitrogen and amino acid true cecal digestibility was high for both T- and T+++ diets (> 90%), but a tendency toward higher nitrogen digestibility was observed for the T- diet (96.6 ± 3.1%) compared to the T+++ diet (91.9 ± 5.4%, P=0.0891). This slightly increased digestibility led to a greater increase in total amino acid concentration in plasma after ingestion of the T- diet (P=0.0010). Comparable protein quality between the two infant formulas was found with a Digestible Indispensable Amino Acid Score (DIAAS) of 0.8. In conclusion, this study showed that minimal processing routes to produce native infant formula do not modify protein quality but tend to enhance its true nitrogen digestibility and increase postprandial plasma amino acid kinetics in rats.
ano.nymous@ccsd.cnrs.fr.invalid (Juliane Calvez) 30 Nov 2023
https://hal.inrae.fr/hal-04315650
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[hal-04299538] The structure of the food matrix at different length scales affects the mechanisms of digestion and the nutrient bioavailability
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ano.nymous@ccsd.cnrs.fr.invalid (Didier Dupont) 22 Nov 2023
https://hal.inrae.fr/hal-04299538
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[hal-04299256] New Insights into Host-Pathogen Interactions of Staphylococcus aureus: Involvement of Extracellular Vesicles to Manipulate Host Cells
Introduction and objectives Like almost all living cells, Staphylococcus aureus produced extracellular vesicles (EVs), which are composed of a lipid bilayer protecting a large number of bioactive molecules such as proteins, lipids, metabolites and nucleic acids. In the past years, there has been growing interest for EVs of bacteria due to their roles in infection and pathogenesis. Indeed, many studies demonstrate their involvement in cell-to-cell communication by transporting and transferring their cargo to recipient cells. EVs may have distinct activities, depending on the producing cell, their functional cargo and their mode of action in recipient cells. It appears that EVs must be central in host-pathogen interactions. However, researches on how pathogen derived-EVs act on host cells are still scarce. Materials and methods Our first objective was to provide a physico-biochemical characterization EVs produced by the methicillin-resistant Staphylococcus aureus strain N315. To mimic infection, N315 strain was grown in RPMI + 10% LB medium and N315-derived EVs were purified by size exclusion chromatography from culture supernatants. EV size and concentration were determined by nanoparticle tracking analysis. Western blot with anti-lipoteichoic acids and anti-peptidoglycan antibodies were performed on EVs lysate sample. Whole-cell and EVs proteome was identified by LC-MS. EVs was treated with RNAse A to remove external RNAs contaminant and the RNA content was analyzed with Bioanalyzer. The second objective was to evaluate impacts of EVs on the expression of several inflammatory genes, as well as their routes of entry into human non-phagocytic cells. EVs uptake by the human osteoblast-like MG- 63 non-phagocytic cell line was evaluated by confocal microscopy and flow cytometry in presence of various transport inhibitors. The impact of EVs on the expression of several MG-63 immune related genes was determined by RT-qPCR. Results, discussion and conclusion S. aureus N315 produced EVs of a typical spherical shape with an average size around 90 nm. Our results show that EVs contained lipoteichoic acid, peptidoglycan and ribonucleic acid. Proteomic comparison of whole cells and its derived EVs revealed the selective packing mechanisms of proteins into N315 EVs, notably of lipoproteins. MG-63 cells can internalize EVs in a dose- and time-dependent manner and via mainly dynamin-mediated endocytosis. EVs induced the expression of numerous immune and signaling genes such as IL-6, IL-8, MCP-1, IL1 , TLR2, TLR4 and TLR7. The expression of some them drastically decreased when EVs uptake was blocked, while others remained unchanged. Altogether, our results showed the ability of N315 EVs to trigger both extracellular and intracellular signaling pathway in host cells. This works shed light on new insights into staphylococcal pathogenesis and potentially provide new alternatives for the treatment and prevention of S. aureus infections.
ano.nymous@ccsd.cnrs.fr.invalid (Julia Papail) 22 Nov 2023
https://hal.inrae.fr/hal-04299256
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[hal-04295498] Extracellular vesicles from Staphylococcus aureus remodulate the bovine mammary epithelial cell transcriptome
Introduction and objectives Staphylococcus aureus is an important etiological agent of mastitis, a disease that affects dairy herds causing great economic losses worldwide. Treatment against S. aureus is still ineffective, and its pathogenicity is not yet fully understood. Secretory factors are important components in bacterial pathogenesis, and the secreted nanosized membrane particles, also known as extracellular vesicles (EVs), play a crucial role in intercellular communication. It has been shown that EVs secreted by S. aureus Newbould 305 strain (NWB305), a bovine mastitis isolate, promote cytokine production in vitro, and tissue inflammation and deterioration in vivo, suggesting their role in mastitis pathogenesis. Here, we investigate how NWB305 EVs interact with bovine mammary epithelial cells (bMECs). Materials and methods RNA-seq approach was used to assess bMECs genes modulated by EVs (3 h, 4 h, 8 h and 24 h) and live NWB305 (3 h) in vitro. Transcriptomic results were validated in vivo by RT-qPCR of mice mammary gland tissues. EV uptake by bMECs was evaluated by confocal microscopy and flow cytometry in presence of various transport inhibitors. Results, discussion and conclusion The number of bMECs differentially expressed (DE) genes modulated by EVs increased over time, being 161 modulated at all conditions that included important cytokines and inflammatory genes (eg. NF-κB, IL-6, IL-8, IL-1β). Interestingly, common genes modulated by both EVs and NWB305 after 3 h comprised those with similar and different expression levels, depending on the source of stimulation. EVs also exclusively modulated 125 genes at 3 h, some of which are involved in apoptosis (eg. APOL-3). Finally, approximately half of the genes modulated by EVs at 24 h were also modulated by NWB305 at 3 h, suggesting that EVs stimulation is slower. The modulation of some of these genes by EVs and NWB305 was confirmed in mice mammary glands in vivo. Altogether, our data show that NWB305 EVs may modulate the host immune response with different intensities, exposure times, and by different routes of those of bacteria in different models. Our study uncovers new EV mechanisms of action and potentially functional roles, providing new insights on their impact in staphylococcal pathogenesis.
ano.nymous@ccsd.cnrs.fr.invalid (Brenda Silva Da Luz) 20 Nov 2023
https://hal.inrae.fr/hal-04295498
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[hal-04286431] Integration of metataxonomic datasets into microbial association networks highlights shared bacterial community dynamics in fermented vegetables
The management of food fermentation is still largely based on empirical knowledge, as the dynamics of microbial communities and the underlying metabolic networks that produce safe and nutritious products remain beyond our understanding. Although these closed ecosystems contain relatively few taxa, they have not yet been thoroughly characterized with respect to how their microbial communities interact and dynamically evolve. However, with the increased availability of metataxonomic datasets on different fermented vegetables, it is now possible to gain a comprehensive understanding of the microbial relationships that structure plant fermentation.
ano.nymous@ccsd.cnrs.fr.invalid (Romane Junker) 15 Nov 2023
https://hal.inrae.fr/hal-04286431
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[hal-04299354] An INFOGEST international consensus static in vitro digestion model adapted to the general older adult population and its application to dairy products
Understanding the mechanisms of food digestion is of paramount importance to determine the effect foods have on human health. Significant knowledge on the fate of food during digestion has been generated in healthy adults due to the development of physiologically-relevant in vitro digestion models. However, it appears that the performance of the oro-gastrointestinal tract is affected by ageing and that a model simulating the digestive conditions found in a younger adult (<65 y) is not relevant for an older adult (>65 y). The objectives of this work were: (1) to conduct an exhaustive literature search to find data on the physiological parameters of the older adult oro-gastrointestinal tract, (2) to define the parameters of an in vitro digestion model adapted to the older adult, (3) to apply it to the digestion of dairy products. International experts have discussed all the parameters during a dedicated workshop organized within the INFOGEST network. Data on food bolus properties collected in the older adult were gathered, including food particle size found in older adult boluses. In the stomach and small intestine, data suggest that significant physiological changes are observed between younger and older adults. In the latter, the rate of gastric emptying is slowed down, the pH of the stomach content is higher, the amount of secretions and thus the hydrolytic activities of gastric and intestinal digestive enzymes are reduced and the concentration of bile salts lower. The consensus in vitro digestion model of the older adult was applied to the digestion of 2 fermented dairy products formulated with a ratio of whey proteins to caseins of 80/20 and 20/80. Results showed that the digestion conditions used (young vs. older adult) influenced significantly the kinetics and extent of proteolysis in the gastric phase but not in the intestinal phase.
ano.nymous@ccsd.cnrs.fr.invalid (Didier Dupont) 22 Nov 2023
https://hal.inrae.fr/hal-04299354
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[hal-04294362] Quantifying lipolysis with MRI during in vitro digestion of cream
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ano.nymous@ccsd.cnrs.fr.invalid (Ruoxuan Deng) 19 Nov 2023
https://hal.inrae.fr/hal-04294362
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[hal-04272108] La qualité des ingrédients protéiques laitiers impacte la physiologie intestinale et le microbiote d’un modèle animal du nourrisson
Les préparations pour nourrissons (PPNs), seul substitut adéquat du lait humain, sont des matrices complexes qui nécessitent de nombreux ingrédients et étapes de transformation. Précédemment, nous avons montré que la qualité des ingrédients protéiques laitiers des PPNs modulaient leur microstructure, leurs cinétiques digestives in vitro et in vivo. L'objectif était donc d’évaluer l'impact de la qualité des protéines laitières sur la physiologie intestinale et le microbiote. Trois PPNs contenant des protéines du lactosérum avec différents niveaux de dénaturation (PPNs-A/-B) ou des caséines d’organisation différentes (PPNs-B/-C) ont été fabriquées. Vingt-quatre mini-porcelets Yucatan, ont reçu l'une des trois PPNs entre 2 et 21 jours d’âge. Les contenus et tissus digestifs ont été analysés via des approches métagénomique, histologique, de perméabilité ex vivo et d’expression génique. Des analyses statistiques univariée et multivariée ont été réalisées. La perméabilité paracellulaire colique était augmentée et un léger boost immunitaire était observé pour les porcelets nourris avec la PPN-C par rapport à la PPN-A, en faveur de la PPN-C, comparativement aux données d’une étude récente sur le lait humain chez le porcelet (Charton et al., 2022). L’expression colique de certains transporteurs d'acides aminés était diminuée chez les porcelets nourris avec la PPN-C par rapport à la PPN-A. La composition du microbiote était similaire selon les régimes et seule la composition fécale en acides gras libres différait selon l’origine du lactosérum dans les PPNs. L’utilisation de lactosérum idéal et la modulation de l’organisation des caséines semblent se présenter comme des pistes d’améliorations pour les PPNs.
ano.nymous@ccsd.cnrs.fr.invalid (Lucile Chauvet) 06 Nov 2023
https://hal.inrae.fr/hal-04272108
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[hal-04272195] Qualité nutritionnelle des protéines de deux coproduits bovins déterminée chez le porc en croissance.
La demande croissante en aliments et notamment en protéines conduit à la recherche de sources alternatives de protéines. La valorisation de coproduits, comme les coproduits carnés peu valorisés dans l’alimentation humaine, est une voie prometteuse pour relever ce défi, qui s’inscrit par ailleurs dans le principe d’économie circulaire. La présente étude visait à évaluer la qualité nutritionnelle de deux ingrédients protéiques d’origine bovine (Greasy Greaves Recovered Proteins-GGRP et Water Recovered Proteins-WRP), tous deux coproduits du processus de fonte des graisses. Leur digestibilité iléale vraie (TID), leur score d'acides aminés indispensables digestibles (DIAAS) et leur cinétique des acides aminés plasmatiques (AAs) ont été mesurés sur modèle animal à partir de prélèvement de digesta et de sang dans les 9 heures suivant l'ingestion des repas. Les TID du N total de WRP et GGRP étaient supérieures à 80%. Le principal AA limitant était le tryptophane pour les deux ingrédients protéiques, avec un DIAAS beaucoup plus élevé pour le GGRP (74%) que pour le WRP (18%) chez la population >3ans. La concentration plasmatique postprandiale d'AA atteignant sa valeur maximale plus tôt pour le WRP (3h) que pour le GGRP (5h). La concentration plasmatique des AA totaux et essentiels était plus élevée avec le repas GGRP qu'avec le repas WRP, alors que l'inverse était vrai pour l'urée. Le GGRP a une qualité nutritionnelle adaptée pour répondre aux besoins de la population >3ans, tandis que le WRP doit être complété par d'autres sources de protéines riches en AAs essentiels pour répondre aux besoins alimentaires.
ano.nymous@ccsd.cnrs.fr.invalid (Rozenn Le Foll) 06 Nov 2023
https://hal.inrae.fr/hal-04272195
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[hal-04281660] In vitro digestion of protein-rich dairy products in the ageing gastrointestinal tract
Aim: Insufficient protein intake can lead to a condition called sarcopenia, characterized by the loss of muscle mass, strength and function. To avoid this condition healthy older adults need to increase the amount of high-quality proteins in their diet, 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 digestion of two high-protein (10% w/w) dairy products in vitro: a fermented dairy product formulated with a ratio of whey proteins to caseins of 80 to 20% (WBD), and a Skyr containing mainly caseins. Method: The new static in vitro digestion model adapted to the general older adult population (≥65 y) proposed by the INFOGEST international consortium was implemented to investigate the digestion of these products, and compared to the standard version of the protocol. Kinetics of proteolysis were compared between both models for each product, in the gastric and intestinal phase of digestion. Protein hydrolysis was studied with the OPA method, SDS-PAGE, and LC-MS/MS, and amino acids were quantified by HPLC. Results: Protein hydrolysis by pepsin was slower with the older adult model than with the young adult model, and consequently, in spite of a longer gastric phase duration, the degree of proteolysis (DH) reached at the end of the gastric phase was lower. Two different scenarios were observed depending on the type of dairy product studied: -10 and -40% in DH for Skyr and WBD, respectively. In the intestinal phase, lower concentrations in free leucine were observed in older adult conditions (approx. -10%), but no significant differences in proteolysis were measured overall between models. Conclusion: The digestion conditions used influenced significantly the kinetics and extent of proteolysis in the gastric phase but not in the intestinal phase. A clinical study comparing the effect of WBD or Skyr on older adults’ postprandial muscle synthesis is underway, which should improve our understanding of differences in digestion between whey protein and casein-based products.
ano.nymous@ccsd.cnrs.fr.invalid (Anaïs Lavoisier) 13 Nov 2023
https://hal.inrae.fr/hal-04281660
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[hal-04282546] Microencapsulation of Mānuka essential oil by plant and animal protein mix systems
Plant proteins are emerging as a valuable dietary resource in contemporary nutrition due to their high protein content and cost-effectiveness. Legumes, prominent among plant sources, are not only protein-rich but also serve as staple food in various global cultures. Nonetheless, it is essential to note that legume proteins are deficient in specific essential amino acids. Consequently, integrating both plant and animal proteins into the formulation of protein mix systems becomes crucial. These systems play a pivotal role in safeguarding and delivering bioactive ingredients. Hence, this study was undertaken to assess the efficacy of plant and animal protein mix systems as potential encapsulation materials for mānuka essential oil. Additionally, an investigation into the physicochemical characteristics and the in vitro release profile of the encapsulated mānuka essential oil was studied. The microcapsules containing mānuka essential oil exhibited minimal moisture content and low water activity. The alteration in the ratio of wall material had a notable impact on the color of the microcapsules. These results revealed the effective encapsulation of mānuka essential oil by the protein mix systems, showcasing controlled release and heightened oxidative stability. This suggests the suitability of these protein mix systems as carriers in encapsulation processes, with significant application potential in the development of functional foods with encapsulated benefits.
ano.nymous@ccsd.cnrs.fr.invalid (Kandi Sridhar) 13 Nov 2023
https://hal.inrae.fr/hal-04282546
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[hal-04273535] Functional Swiss-type cheeses promote beneficial effects in mice gut microbiome during homeostasis and inflammation
The impact of Functional foods on the gut microbiota is a fundamental question since shifts in bacterial composition are associated with inflammation progression. Therefore, this study investigated the effects of two types of cheeses fermented by probiotic bacteria in C57BL6 mice either in healthy conditions or in dextran sulfate sodium (DSS)-induced colitis. One cheese was fermented by Propionibacterium freudenreichii CIRM-BIA129. A second kind of cheese was made in industrial conditions to reproduce Emmental but using the above-mentioned strain in combination with Lactobacillus delbrueckii CNRZ327 and Streptococcus thermophilus LMD-9 as starters. The gut microbiota was investigated by shotgun metagenomic sequencing and the taxonomic and functional profiles were assessed through sequence alignment to NCBI taxonomy and KEGG pathways databases. Our results suggest that both treatments did not disturb the microbial community ecology. Emmental intake increased symbionts as Romboutsia and Akkermansia muciniphila. Furthermore, metabolic pathway analysis suggests that A. muciniphila may produce bioactive metabolites as acetate and cooperate with other commensal species to produce indole and gamma-aminobutyric acid. Mice with colitis have restored some of the impaired microbiome metabolic functions when treated with cheese fermented only by Propionibacterium, while the Emmental cheese promoted the increase of Ligilactobacillus murinus. This taxon presented several genes with immunomodulatory activity potential. This study provides insights for engineering functional cheeses to increase beneficial bacteria in the gut. However, further studies are required to investigate their role in regulating the gut-brain axis. Moreover, possible anti-inflammatory mechanisms based on the promotion of Lg. murinus interaction with the host was suggested.
ano.nymous@ccsd.cnrs.fr.invalid (Rodrigo Dias de Oliveira Carvalho) 07 Nov 2023
https://hal.inrae.fr/hal-04273535
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[hal-04260177] A proteomic analysis of immunomodulatory effects mediated by the probiotic propionibacterium freudenreichii and by its EVs
[...]
ano.nymous@ccsd.cnrs.fr.invalid (Gwénaël Jan) 26 Oct 2023
https://hal.inrae.fr/hal-04260177
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[hal-04257450] Interaction mechanisms with host of EVs derived from the opportunistic pathogen Staphylococcus aureus
[...]
ano.nymous@ccsd.cnrs.fr.invalid (Julia Papail) 25 Oct 2023
https://hal.inrae.fr/hal-04257450
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[hal-04254337] Health Benefits of EVs Derived from the Probiotic Bacterium Propionibacterium freudenreichii
[...]
ano.nymous@ccsd.cnrs.fr.invalid (Juliana Laguna) 23 Oct 2023
https://hal.inrae.fr/hal-04254337
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[hal-04253707] Research and Innovation in Nutrition : A major health issue Milk products and innovative fermented ingredients for target populations : Focus on the PROLIFIC Project
Prolific : étudier le potentiel santé des bactéries lactiques Le projet de recherche Prolific (Produits laitiers et ingrédients fermentés innovants pour des populations cibles) vise à évaluer les bénéfices santé des produits laitiers fermentés chez les jeunes enfants et les seniors. Le projet porté par un consortium associant des industriels regroupés au sein de Bba Milk Valley® et des équipes de recherche bretonnes et ligériennes va nécessiter un investissement de 14 millions d'euros sur 5 ans. Yves Le Loir, directeur de l’UMR STLO, est chargé de coordonner le projet. Partenaires : UMR STLO, UMR NuMeCan, UMR TENS, INRA BIA, IRISA, UMR Combi LS2N, Bba.
ano.nymous@ccsd.cnrs.fr.invalid (Yves Le Loir) 23 Oct 2023
https://hal.inrae.fr/hal-04253707
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[hal-04249944] Fat matters: fermented whole milk potentiates the anti-colitis effect of Propionibacterium freudenreichii
Inflammatory bowel diseases (IBD) constitute a growing concern in western countries. They coincide with gut microbiota dysbiosis, including a loss of immunomodulatory bacteria. Accordingly, probiotic products containing selected immunomodulatory bacterial strains mitigate IBD. Selected strains of Propionibacterium freudenreichii display promising modulatory properties and prevent colitis in animal models. This presentation will summarise the research work done on P. freudenreichii preventive potential in the context of colitis and of mucositis. Dairy matrices protect propionibacteria immunomodulatory surface antigens during digestive transit. However, the functional role of the dairy matrix components in such fermented dairy products remains unknown. P. freudenreichii CIRM-BIA129, a probiotic strain known for its anti-inflammatory properties, was used to ferment whole milk, skim milk or skim milk ultrafiltrate. The preventive potential of fermented products was tested in DSS-induced mice colitis, in comparison with their unfermented counterparts. P. freudenreichii-fermented milk prevented colitis. Dairy fat in the fermented product potentiated the anti-colitis effects of the probiotic. This work opens new perspectives for developing immunomodulatory functional fermented foods.
ano.nymous@ccsd.cnrs.fr.invalid (Marine Mantel) 19 Oct 2023
https://hal.inrae.fr/hal-04249944
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[hal-04251786] Unveiling the Secrets of Long-Term Staphylococcus aureus Infections: Insights into Inflammation, Metabolism, and Epigenetic Changes from Non-Immune Cells Transcriptomes
Staphylococcus aureus, an opportunistic pathogen, is responsible for a spectrum of severe chronic diseases such as chronic osteomyelitis and chronic mastitis in humans and in cows, respectively. Although traditionally regarded as an extracellular pathogen, S. aureus has been identified within non-immune cells such as osteoblasts (associated with osteomyelitis) or mammary gland epithelial cells (linked to cow mastitis), potentially influencing the infection outcome. The detection of signals specific from infected bacteria-bearing cells is complicated by dilution and confounding signals by bystander effects in uninfected cells. To address this, we developed a novel long-term infection model for non-immune cells using a flow cytometric approach, enabling the isolation of cells containing internalized S. aureus from mixed populations. This model facilitates a detailed analysis of the impact of prolonged S. aureus infection on the transcriptional program of human non-immune (osteoblast-like) cells. RNA-seq, KEGG, and Reactome pathway enrichment analyses revealed a remodeled transcriptomic profile marked by heightened immune and inflammatory responses, metabolic dysregulations influencing bacterial intracellular life, and downregulation of genes encoding epigenetic regulators, including those involved in chromatin-repressive complexes (e.g., NuRD, BAHD1, and PRC1) and epifactors associated with DNA methylation. Additionally, sets of genes related to cell adhesion or neurotransmission were deregulated. Our findings suggest a long-term impact of intracellular S. aureus infection through epigenetic modifications in host cells, potentially leading to pathophysiological dysfunctions alongside the defense response during infection. These results not only enhance our understanding of biological processes in long-term S. aureus infections of non-immune cells but also provide an atlas of deregulated host genes and biological pathways, identifying potential markers and candidates for prophylactic and therapeutic approaches. Moreover, building on this in vitro work, we have initiated a collaboration with the NuMeCan Institute to validate these observations in vivo in a mini-pig model of S. aureus infection. This animal study will be complemented by an exploration of the serum metabolome and the intestinal microbiota in the chronicity of S. aureus infection. Furthermore, these investigations will be extended to the context of S. aureus mastitis in cows through collaboration with Canadian colleagues.
ano.nymous@ccsd.cnrs.fr.invalid (Aurélie Nicolas) 20 Oct 2023
https://hal.inrae.fr/hal-04251786
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[hal-04294381] Monitoring the effect of consumption temperature of whole fat milk on in vitro gastric digestion using magnetic resonance imaging
[...]
ano.nymous@ccsd.cnrs.fr.invalid (Conor Fitzpatrick) 19 Nov 2023
https://hal.inrae.fr/hal-04294381
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[hal-04251333] Effet de régimes contrastés sur les concentrations en vitamines B des produits laitiers de vache.
Les vitamines présentes dans le lait des ruminants ont été très peu étudiées jusqu’à la fin du XXème siècle. De nouvelles connaissances ont été acquises récemment sur leurs facteurs de variation. L’évolution des conditions d’élevage pour répondre aux problématiques liées aux aléas climatiques, à la transition agroécologique, à l’évolution des habitudes alimentaires et aux souhaits d’augmentation de la qualité nutritionnelle des produits des consommateurs renforce l’intérêt d’approfondir le sujet et de compléter les lacunes.
ano.nymous@ccsd.cnrs.fr.invalid (Lucie Auzance) 20 Oct 2023
https://hal.inrae.fr/hal-04251333
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[hal-04282590] In vitro digestion of protein-rich dairy products in the ageing gastrointestinal tract
Interest for specific populations – Older adults Insufficient protein intake can lead to sarcopenia, characterized by the loss of muscle mass, strength, and function. • Healthy older adults (> 65 y.) need to increase the amount of high-quality proteins in their diet (1 g protein /kg body weight /day). ESPEN guidelines • Ageing leads to changes in the functionality of the digestive tract but the impact of ageing on digestion, and absorption of nutrients is still unclear (Lee et al., 2021). Investigate high-protein dairy products digestion in vitro Study the influence of age on the kinetics of proteolysis in the gastric and intestinal phase of digestion
ano.nymous@ccsd.cnrs.fr.invalid (Anaïs Lavoisier) 13 Nov 2023
https://hal.inrae.fr/hal-04282590
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[hal-04243643] Eco-conception des procédés agro-alimentaires Approches et nouvelles opportunités
L'éco-conception des procédés agro-alimentaires vise à améliorer leurs performances environnementales, technologiques et socio-économiques tout en garantissant la qualité globale des aliments. Cet exposé présentera, d’une part, les démarches existantes d’éco-conception de procédés agro-alimentaires et, d'autre part, les approches émergentes et prometteuses. Ces approches seront illustrées par des exemples tirés d’activités de recherches INRAE et feront l'objet d'une discussion explicite sur les avantages et les défis potentiels de leur application
ano.nymous@ccsd.cnrs.fr.invalid (Geneviève Gésan-Guiziou) 16 Oct 2023
https://hal.inrae.fr/hal-04243643
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[hal-04243750] Development of an ecodesign framework for food manufacturing including process flowsheeting and multiple-criteria decision-making: Application to milk evaporation
The food industry urgently needs to design sustainability into its processes. Here we developa methodological framework for the ecodesign of food processes. The framework combinesprocess flowsheeting (using a process simulation tool) and multiobjective optimization withlife-cycle and cost-cycle analysis tools embedded in a multicriteria decision support tool.The framework developed is illustrated by the example of milk evaporation, which is oneof the most energy-intensive processes in the dairy industry and has huge potential foroptimization. The commercial process simulator was first adapted to modeling milk evapo-rators. Economic and environmental criteria were then computed from inventories of boththe production and cleaning phases, and we went on to apply multiobjective optimization(with a genetic algorithm) and multicriteria decision-making either independently or incombination. The potential of the framework was demonstrated first through analysis ofthree evaporator design solutions, and second as a support for the strategic choice of a fuelfor the purpose of on-site energy production. This work provides a pathway to ecodesigningthe entire milk powder production chain.
ano.nymous@ccsd.cnrs.fr.invalid (Catherine Azzaro-Pantel) 16 Oct 2023
https://hal.inrae.fr/hal-04243750
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[hal-04236644] Les bactéries du lait maternel humain, individuellement ou assemblées en communautés synthétiques, ont un impact sur les fonctions immunitaire et barrière intestinales, in vitro.
L'allaitement confère de nombreux bienfaits pour la santé, en favorisant le développement du système immunitaire et de la fonction barrière de l’intestin chez les nourrissons. Notre hypothèse est que le microbiote du lait maternel humain contribue à ces bénéfices santé. Notre objectif était d'explorer le rôle des bactéries du lait maternel, seules ou assemblées dans des communautés bactériennes synthétiques (SynCom), sur l'homéostasie intestinale. Une collection de bactéries, reflétant la composition du microbiote du lait, a été constituée à partir de 28 donneuses en bonne santé, allaitant exclusivement. Tout d'abord, le profil immunomodulateur de 84 souches représentatives de la diversité taxonomique du lait maternel a été étudié sur des cellules mononucléées du sang périphérique (PBMC). Ensuite, un modèle quadricellulaire (Caco2, HT29-MTX-E12, cellules M, THP1) de l'épithélium intestinal a été utilisé pour caractériser plus finement un sous-ensemble de 29 souches. Les isolats ont été caractérisés pour leur capacité à moduler la production d'IL-10 et de TNF-α et l'expression de gènes associés notamment aux fonctions barrière et immunitaire. Sur la base de ces résultats, deux SynComs ont été créées et caractérisées sur le modèle quadricellulaire. Les analyses intégratives « MultiDimensional Scaling » et sPLS-DA (sparse Partial Least Squares-Discriminant Analysis) des variables mesurées sur le modèle PBMC puis sur le modèle quadricellulaire stimulé avec les souches du lait maternel ont révélé des groupes de souches ayant un impact différent sur la production de cytokines, l'expression de gènes impliqués dans l'immunité, la fonction barrière et le signaling bactérien. Ces groupes fonctionnels sont très faiblement corrélés à la taxonomie et leur composition souligne l’intérêt de genres et d’espèces prévalents du lait maternel (Staphylococcus, Cutibacterium…) au même titre que des genres plus explorés dans ce contexte (Bifidobacterium, Lactobacillus). Enfin, les souches assemblées dans deux SynComs représentatives du microbiote du lait (11 souches) ont présenté des propriétés immunomodulatrices contrastées. Cette étude montre la grande diversité de potentiel immunomodulateur et d'impact sur la fonction de barrière des bactéries du lait maternel, individuellement ou assemblées en SynCom, soulignant le potentiel de ce microbiote pour moduler le développement de l'homéostasie intestinale.
ano.nymous@ccsd.cnrs.fr.invalid (Charles Le Bras) 11 Oct 2023
https://hal.inrae.fr/hal-04236644
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[hal-04234356] Development of innovative fermented products by exploiting the diversity of immunomodulatory properties and fermentative activity of lactic and propionic acid bacteria
Ce projet a été sélectionné suite à l’appel à projet du Fond Unique Interministériel, FUI 25
ano.nymous@ccsd.cnrs.fr.invalid (Gwénaël Jan) 10 Oct 2023
https://hal.inrae.fr/hal-04234356
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[hal-04234337] Extracellular vesicles from Staphylococcus aureus follow different pathways to manipulate host cells
Introduction and objectives Extracellular vesicles (EVs) are nanosized membrane-encapsulated particles produced by almost all living cells and loaded with various molecules (eg, lipids, nucleic acids, proteins). They play important roles in cell-to-cell communication by transporting and transferring their cargo to recipient cells. EVs may have distinct activities, depending on the producing cell, their functional cargo and their mode of action in recipient cells. EVs produced by bacterial pathogens contribute to pathogenicity as mediators of host-pathogen interactions. However, how pathogen derived-EVs act on host cells is still poorly documented. Here, we characterized EVs produced by the methicillin-resistant Staphylococcus aureus strain N315, and evaluated their impact on the expression of several inflammatory genes, as well as their routes of entry into human non-phagocytic cells. Materials and methods To mimic infection, N315 strain was grown in RPMI + 10% LB medium and N315-derived EVs were purified by size exclusion chromatography from culture supernatants. EV size and concentration were determined by nanoparticle tracking analysis. Whole-cell and EV proteome was identified by LC-MS. EV uptake by the human osteoblast-like MG-63 non-phagocytic cell line was evaluated by confocal microscopy and flow cytometry in presence of various transport inhibitors. The impact of EVs on the expression of several MG-63 immune related genes was determined by RT-qPCR. Results, discussion and conclusion S. aureus N315 produced EVs of a typical spherical shape with an average size around 90 nm. Proteomic comparison of whole cells and its derived EVs revealed the selective packing mechanisms of proteins into N315 EVs, notably of lipoproteins. In addition, EVs contained lipoteichoic acid, peptidoglycan and ribonucleic acid. MG-63 cells can internalize EVs in a dose- and time-dependent manner and via mainly dynamin-mediated endocytosis. EVs induced the expression of numerous immune and signaling genes such as IL-6, IL-8, MCP-1, IL1B, TLR2, TLR4 and TLR7. The expression of some them drastically decreased when EV uptake was blocked, while others remained unchanged. Altogether, our results showed the ability of N315 EVs to trigger both extracellular and intracellular signaling and via different bacterial components, bringing new insights into staphylococcal pathogenesis.
ano.nymous@ccsd.cnrs.fr.invalid (Julia Papail) 10 Oct 2023
https://hal.inrae.fr/hal-04234337
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[hal-04234619] Extracellular vesicles from Staphylococcus aureus remodulate the bovine mammary epithelial cell transcriptome
Introduction and objectives Staphylococcus aureus is an important etiological agent of mastitis, a disease that affects dairy herds causing great economic losses worldwide. Treatment against S. aureus is still ineffective, and its pathogenicity is not yet fully understood. Secretory factors are important components in bacterial pathogenesis, and the secreted nanosized membrane particles, also known as extracellular vesicles (EVs), play a crucial role in intercellular communication. It has been shown that EVs secreted by S. aureus Newbould 305 strain (NWB305), a bovine mastitis isolate, promote cytokine production in vitro, and tissue inflammation and deterioration in vivo, suggesting their role in mastitis pathogenesis. Here, we investigate how NWB305 EVs interact with bovine mammary epithelial cells (bMECs). Materials and methods RNA-seq approach was used to assess bMECs genes modulated by EVs (3 h, 4 h, 8 h and 24 h) and live NWB305 (3 h) in vitro. Transcriptomic results were validated in vivo by RT-qPCR of mice mammary gland tissues. EV uptake by bMECs was evaluated by confocal microscopy and flow cytometry in presence of various transport inhibitors. Results, discussion and conclusion The number of bMECs differentially expressed (DE) genes modulated by EVs increased over time, being 161 modulated at all conditions that included important cytokines and inflammatory genes (eg. NF-κB, IL-6, IL-8, IL-1β). Interestingly, common genes modulated by both EVs and NWB305 after 3 h comprised those with similar and different expression levels, depending on the source of stimulation. EVs also exclusively modulated 125 genes at 3 h, some of which are involved in apoptosis (eg. APOL-3). Finally, approximately half of the genes modulated by EVs at 24 h were also modulated by NWB305 at 3 h, suggesting that EVs stimulation is slower. The modulation of some of these genes by EVs and NWB305 was confirmed in mice mammary glands in vivo. Altogether, our data show that NWB305 EVs may modulate the host immune response with different intensities, exposure times, and by different routes of those of bacteria in different models. Our study uncovers new EV mechanisms of action and potentially functional roles, providing new insights on their impact in staphylococcal pathogenesis.
ano.nymous@ccsd.cnrs.fr.invalid (Brenda Silva Rosa da Luz) 10 Oct 2023
https://hal.inrae.fr/hal-04234619
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[hal-04257483] Ability of Lactic acid bacteria to ferment cheese whey to produce whey-based drinks
Cheese whey is the watery part obtained after milk curdling in cheese-making. With up to 9 L obtained for 1 kg of cheese produced, whey constitutes a major by-product of dairy industry. It mostly contains water but also lactose, minerals, vitamins (B6, B1, B12) and high-quality proteins. Despite these obvious nutritional qualities, the sour and salty taste of whey makes it not appealing to the consumer and restricts its direct consumption or use in food. For this reason, whey has long been poorly valorized and mainly used as feedstock for pigs or discarded. Nowadays, valorization rely on whey protein extraction/purification by heavy industrial process to obtain value-added food ingredients. However, these processes are only applicable to some cheese wheys, are energy-consuming and mostly concern large-scale dairy plants leaving small ones out of the system. As part of FAIRCHAIN project, we aimed at developing whey-based drinks, as alternative whey valorization, using microbial fermentation to preserve whey while also improving its native sensory properties. This study explores the ability of Lactic Acid Bacteria (LAB) to ferment and acidify different types of whey and obtain the best aroma profiles for the drinks.
ano.nymous@ccsd.cnrs.fr.invalid (Marine Penland) 25 Oct 2023
https://hal.inrae.fr/hal-04257483
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[hal-04233142] Impacts au niveau microbien, immunitaire et physiologique de l’application post-traite d'une souche de bactérie lactique sur la peau du trayon pour la prévention des mammites bovines
Les mammites bovines (MB) constituent un problème récurrent majeur pour l'industrie laitière. Les antibiotiques utilisés pour traiter les MB ne sont pas totalement efficaces et peuvent contribuer au risque de dissémination de résistances antimicrobiennes. Leur prévention repose donc principalement sur la désinfection des trayons avec des produits chimiques tels que l'iode dont l’utilisation est remise en cause du fait d'une possible irritation de la peau et de la dissémination de résidus dans le lait. Il est donc important de développer des stratégies alternatives de prévention des MB. Cette étude vise à évaluer l’impact de l'application post-traite d’une souche de bactérie lactique (BL) sur la peau du trayon en vue de renforcer l’effet barrière du microbiote naturellement présent sur le trayon. Pour cela, le traitement avec la BL a été appliqué pendant 15 jours sur les quartiers de vaches Holstein (n=8 vaches) et comparé au traitement à l’iode (n=6) et à l’absence de traitement (n=6). Des échantillons de lait, de sang et des écouvillons de peau du trayon ont été prélevés à J1, J8, J15 et J26 pour évaluer l'impact du traitement avec la BL au niveau microbien, immunitaire et physiologique. La BL est présente sur la peau du trayon et dans le lait de 1er jet pendant le traitement et cela s’accompagne d’une augmentation significative de la population microbienne totale. L’analyse métataxonomique du microbiote de ces 2 sites anatomiques indique un faible impact du traitement avec la BL sur leur richesse mais un impact significatif sur leur composition. Le traitement avec la BL n'a déclenché aucune réponse inflammatoire majeure (cellules somatiques et IL-8 dans le lait) et aucun impact négatif significatif sur la fonctionnalité et l'intégrité de l'épithélium mammaire (production et composition du lait, exfoliation des cellules épithéliales, rapport Na+:K+ dans le lait). Ces résultats révèlent un impact du traitement avec la BL sur le microbiote du trayon tout en indiquant l’absence d’effets délétères sur la réponse immunitaire et l’intégrité de l’épithélium mammaire, ouvrant la voie sur l’utilisation de méthodes de prophylaxie plus durables et plus respectueuses de l'environnement, de la santé et du bien-être des animaux et de l’homme.
ano.nymous@ccsd.cnrs.fr.invalid (Coralie Goetz) 09 Oct 2023
https://hal.inrae.fr/hal-04233142
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[hal-04234405] Development of trained immunity and activation of inflammasomes are promising strategies to combat Staphylococcus aureus infection
Introduction and objectives Staphylococcus aureus causes life-threatening diseases such as pneumonia and osteomyelitis. Trained immunity (TI) enhances the immune response to subsequent unrelated challenges through epigenetic reprogramming of transcriptional pathways and alteration of cell metabolism [1]. Moreover, pathogens activate inflammasomes that trigger protease activation, particularly caspase-1. Caspase-1 proteolytically matures and promotes the secretion of mature IL-1β and IL-18 [2]. Our objectives include investigating the development of TI in non-immune cells, examining interleukin production and epigenetic marks, exploring the involvement of reactive oxygen species (ROS), and comprehending the role of inflammasomes and their effector, caspase-1, in the context of S. aureus infection. Materials and methods We developed an in vitro TI model using human osteoblast-like MG-63 and epithelial lung A549 cells [3]. Techniques employed included microbiology methods of bacterial cultivation, enzyme-linked immunosorbent assay (ELISA), microscopic analysis, and flow cytometry. We investigated the involvement of ROS using the scavenger N-acetylcysteine (NAC) [4]. The role of inflammasomes and Caspase-1 was examined by a comparison of MG-63 cells and caspase-1 knock-out CASP1−/−MG-63 cells that were generated via CRISPR-Cas9 technology [5]. The role of bacterial effectors were analyzed using deletion and complemented phenol-soluble modulins (PSMs) mutants. Results, discussion and conclusion β-glucan training of MG-63 and A549 cells increased IL-6/IL-8 production upon a stimulation with S. aureus. Interleukin production positively correlated with Histone 3 acetylation at lysine 27 (H3K27), indicating epigenetic reprogramming. NAC addition, prior to β-glucan training before S. aureus infection, inhibited IL-6/IL-8 production, thereby supporting the involvement of ROS in the induction of TI. Cell exposure to bacterium with probiotic properties, Lactococcus lactis, before S. aureus infection also elevated IL-6/IL-8 production, accompanied by H3K27 acetylation, suggesting its ability to induce TI [3]. Infected MG-63 cells activated inflammasomes, releasing mature IL-1β, while CASP1−/−MG-63 cells lacked this activation. PSMs were identified as key contributors to inflammasome-related IL-1β production using S. aureus mutants. Additionally, caspase-1 deficiency impaired cell defense, resulting in decreased bacterial clearance in CASP1−/−MG-63 cells [5]. Our results demonstrate two host strategies against S. aureus infection with therapeutic potential and reveal the involvement of non-immune cells in defense response beyond their structural functions and tissue homeostasis maintenance.
ano.nymous@ccsd.cnrs.fr.invalid (Emmanuel Chaumond) 10 Oct 2023
https://hal.inrae.fr/hal-04234405
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[hal-04233228] Les métabolites bactériens naturellement présents dans le lait maternel possèdent des propriétés bioactives : étude in vitro dans un modèle quadricellulaire d’épithélium intestinal et un modèle de neurones centraux.
Le lait maternel (LM) est un aliment complexe contenant de nombreux éléments dont des métabolites. Ceux-ci sont d’origine maternelle ou peuvent être synthétisés par les bactéries naturellement présentes dans le LM. C’est le cas des acides gras à chaîne courte (AGCC) (butyrate, acétate, propionate), des polyamines (putrescine, spermine, spermidine), du GABA et du lactate, qui sont retrouvés à faible concentration dans le LM. Ces métabolites sont connus pour être produits par le microbiote intestinal et être des acteurs de l’axe microbiote-intestin-cerveau, mais leur rôle au sein du LM a été peu étudié. L’objectif de cette étude a été d’évaluer les propriétés bioactives de ces métabolites bactériens sur les fonctions intestinales et cérébrales du nouveau-né. Ces métabolites ont été étudiés in vitro, à des concentrations proches de celles du LM, dans un modèle pluricellulaire et polarisé d’épithélium intestinal composé de Caco-2 (entérocytes), de HT-29-MTX (cellules à Gobelet), de NCI-H716 (cellules entéroendocrines) et de cellules M, ainsi que dans un modèle de neurones centraux. Les effets des métabolites sur la résistance électrique transépithéliale (TEER) et l'expression de marqueurs géniques ou protéiques des fonctions intestinales et cérébrales ont été analysés. Les AGCC ont agi sur des marqueurs de l’immunité intestinale en régulant négativement les gènes codant pour l'IL-8, MyD88 et TFF3. Ils ont également modulé l’expression de certaines jonctions serrées intestinales, tout comme le GABA et les polyamines, en augmentant l’expression de CLDN3, ZO-1 et CLDN4 respectivement et en diminuant celle de CLDN1 pour les AGCC et les polyamines et CLDN7 pour le GABA. En parallèle, les AGCC ont significativement augmenté la TEER, mettant en évidence un potentiel renforcement sur la barrière épithéliale. Les AGCC et le GABA ont également modulé certains marqueurs de connectivité et de maturation neuronales (synaptophysine, synapsine, PSD95 et doublecortine). En conclusion, notre étude montre que les métabolites bactériens du LM, malgré leur faible concentration, sont capables de moduler les fonctions intestinales et cérébrales in vitro, suggérant un impact potentiel sur la physiologie et développement du nourrisson.
ano.nymous@ccsd.cnrs.fr.invalid (Sarah Blanchet) 09 Oct 2023
https://hal.inrae.fr/hal-04233228
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[hal-04234937] Value of the Galleria mellonella model for exploring the Staphylococcus aureus extracellular vesicles impact on host-pathogen interactions
Introduction and objectives Extracellular vesicles (EVs) are nanometric particles secreted from the membrane into the surrounding environment. They play a crucial role in intercellular communication through their ability to transport bioactive molecules. Various functions have been attributed to EVs produced by Staphylococcus aureus: biofilm formation, cytotoxicity, immunomodulation... These functions suggest their involvement in host-pathogen interactions and pathogenesis. However, in vivo data remain scarce to date. The aim of this work was to test the larval Galleria mellonella model to assess the impact of EVs in vivo. Materials and methods Derivated HG003 reference strain-EVs were purified by steric exclusion chromatography. Different quantities of EVs were injected into the larval circulatory system to determine the minimum lethal dose. Impact of EVs, alive bacteria and PBS buffer was compared at different times after injection, both on aspects of mortality, macroscopy/microscopy, and expression of factors of larval innate immunity (e.g., opsonins, anti-microbial peptides...) by RT-qPCR from 2 distinct environments, (i) hemolymph (HL; larval circulatory system) and (ii) fat body (FB). All experiments were performed in triplicate. Results, discussion and conclusion Preliminary data showed that only a significant quantity of EVs (1010 EVs) were responsible for early larval mortality. Microscopy data allowed us to visualize S. aureus bacteria as well as clusters of EVs, each condition exhibiting a specific immune response and cellular remodeling. The larval immune response proved to be specific to the condition tested (bacteria vs. EVs), and depended on post-infection time and localization (HL vs. FB). Compared to bacteria, EVs induced an early activation of the majority of tested-innate factors in the HL, whereas a rather late activation in the FB. In addition, certain immune players were specifically induced by EVs. The G. mellonella model therefore appears suitable for assessing the contribution of S. aureus EVs to pathogenesis.
ano.nymous@ccsd.cnrs.fr.invalid (Guillaume Ménard) 10 Oct 2023
https://hal.inrae.fr/hal-04234937
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[hal-04243684] Evaluation MultiCritère des Systèmes Agricoles et Alimentaires (EMC-SAgA) Bilan des activités et Poursuites envisagées
[...]
ano.nymous@ccsd.cnrs.fr.invalid (Geneviève Gésan-Guiziou) 16 Oct 2023
https://hal.inrae.fr/hal-04243684
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[hal-04299393] The structure of the food matrix at different length scales affects the mechanisms of digestion and the nutrient bioavailability
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ano.nymous@ccsd.cnrs.fr.invalid (Didier Dupont) 22 Nov 2023
https://hal.inrae.fr/hal-04299393
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[hal-04246054] In vitro digestion of two protein-rich dairy products in the ageing gastrointestinal tract
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 digestion of two high-protein (10% w/w) dairy products in vitro: a fermented dairy product formulated with a ratio of whey proteins to caseins of 80 to 20% (WBD) and a Skyr containing mainly caseins. The new static in vitro digestion model adapted to the general older adult population (≥65 years) proposed by the INFOGEST international consortium was implemented to investigate the digestion of these products and compared with the standard version of the protocol. Kinetics of proteolysis was compared between both models for each product, in the gastric and intestinal phases of digestion. Protein hydrolysis was studied by the OPA method, SDS-PAGE, and LC-MS/MS, and amino acids were quantified by HPLC. Protein hydrolysis by pepsin was slower with the older adult model than with the young adult model, and consequently, in spite of a longer gastric phase duration, the degree of proteolysis (DH) at the end of the gastric phase was lower. Two different scenarios were observed depending on the type of dairy product studied: −10 and −40% DH for Skyr and WBD, respectively. In the intestinal phase, lower concentrations of free leucine were observed in older adult conditions (approx. −10%), but no significant differences in proteolysis were observed overall between the models. Therefore, the digestion conditions used influenced significantly the rate and extent of proteolysis in the gastric phase but not in the intestinal phase.
ano.nymous@ccsd.cnrs.fr.invalid (Anaïs Lavoisier) 17 Oct 2023
https://hal.inrae.fr/hal-04246054
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[hal-04216723] Design adaptatif de procédés microbiens en système fermé : illustrations pour les aliments.
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ano.nymous@ccsd.cnrs.fr.invalid (Valérie Gagnaire) 25 Sep 2023
https://hal.inrae.fr/hal-04216723
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[hal-04299592] In vitro amino acid digestibility and availability
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ano.nymous@ccsd.cnrs.fr.invalid (Didier Dupont) 22 Nov 2023
https://hal.inrae.fr/hal-04299592
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[hal-04279361] Nutritional quality of proteins from two beef co-products as determined in the growing pig
The increasing demand for food and especially for proteins, leads to the search for alternative protein. The valorization of co-products, such as meat co-products little valorized in human food, is from this point of view a promising way, in line with the principle of circular economy. The present study aimed to evaluate the nutritional quality of two protein ingredients of bovine origin, co-products of the fat rendering process. To do so, their true ileal digestibility (TID), their digestible indispensable amino acid score (DIAAS) and the postprandial plasma amino acid (AA) kinetics were determined. Ten growing pigs were surgically inserted an ileal T-cannula and a catheter in the jugular vein. After recovery, they received each over a 2.5-day period and following a cross-over design two experimental diets based either on Greasy Greaves Recovered Proteins (GGRP) or on Water Recovered Proteins (WRP)], followed by a protein-free diet for endogenous protein loss evaluation. Titanium dioxide (3 g/kg) was used as an undigestible marker. Digesta and blood samples were collected during the 9 hours following meal ingestion and total N and AA contents were analysed by the Dumas method or by ion-exchange chromatography, respectively. Data were statistically analysed using linear models. The TID of total N was not significantly different between WRP and GGRP (84.2±3.2 and 81.2±2.3%, respectively; mean ± SD). The primary limiting AA was tryptophan for both protein ingredients, with a DIAAS much higher for GGRP (74%) than for WRP (18%). The kinetics of AA release differed between proteins, with postprandial plasma AA concentration reaching its maximal value earlier for WRP (3 hours) than for GGRP (5 hours). GGRP has a nutritional quality suitable to meet the needs of old children, adolescents and adults, while WRP should be supplemented with other protein sources rich in essential AAs to fulfill the AA requirements.
ano.nymous@ccsd.cnrs.fr.invalid (Rozenn Le Foll) 10 Nov 2023
https://hal.inrae.fr/hal-04279361
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[hal-04257570] Amino acid requirements for infants
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ano.nymous@ccsd.cnrs.fr.invalid (Amélie Deglaire) 25 Oct 2023
https://hal.inrae.fr/hal-04257570
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[hal-04305637] High protein digestibility and utilisation rate of lamb meat hydrolysate in older adults for healthy ageing
Sarcopenia is an age-related decline in skeletal muscle accompanying low muscle strength and/or low physical performance in older adults (>60 years) with increasing mortality risk1 . The study examines a lamb meat hydrolysate as a rich protein source with enhanced postprandial protein utilisation in older adults.
ano.nymous@ccsd.cnrs.fr.invalid (Sharon J. Henares) 24 Nov 2023
https://hal.science/hal-04305637
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[hal-04206782] Du -80 au digesteur dynamique, le parcours STLO des bactéries d’intérêt alimentaire dans la conception de nouveaux produits laitiers fermentés
Comment concevoir des aliments laitiers fermentés adaptés aux besoins spécifiques de populations cibles ? C’est la question à laquelle s’attèlent les microbiologistes du STLO. Pour cela, ils disposent d’une banque de bactéries parfaitement caractérisées (CIRM-BIA) qu’ils combinent pour former des consortia adaptés aux matrices laitières (et éventuellement végétales) et exprimant des fonctionnalités d’intérêt organoleptique, nutritionnel ou santé. Selon le type de fonctionnalité, les caractéristiques de ces diverses combinaisons sont évaluées par l’analyse des métabolites produits, ou via des tests d’interaction sur culture cellulaire. Ces consortia ainsi élaborés sont d’abord cultivés en conditions de laboratoire sur de petits volumes puis sont mis en œuvre sur des volumes plus importants, en fermenteurs. Les plus prometteurs peuvent ensuite être confrontés aux conditions technologiques réelles lors de fabrications grandeur nature sur notre plateforme de technologie laitière, sous une forme adaptée aux populations cibles (préparation pour nourrissons, yaourt, fromage…). Les produits fermentés obtenus peuvent enfin être soumis aux stress du processus digestif à l’aide de simulateur biomimétique de digestion. Toutes ces étapes permettent de valider les propriétés des consortia d’intérêt et de tester leur comportement en cours de process. Elles génèrent des données hétérogènes. L’ensemble de ces données est encore sous-exploité et pourrait utilement être soumis à des outils de type IA pour élaborer les meilleures combinaisons en réduisant les temps de développements expérimentaux.
ano.nymous@ccsd.cnrs.fr.invalid (Yves Le Loir) 15 Sep 2023
https://hal.inrae.fr/hal-04206782
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[hal-04215082] Involvement of Extracellular Vesicles from Staphylococcus aureus in host cells manipulation
Extracellular vesicles (EVs) are nanosized membrane-encapsulated particles produced by almost all living cells and loaded with various molecules (eg, lipids, nucleic acids, proteins). They play important roles in cell-to-cell communication by transporting and transferring their cargo to recipient cells. EVs may have distinct activities, depending on the producing cell, their functional charge and their mode of action in recipient cells. EVs produced by bacterial pathogens contribute to pathogenicity as mediators of host-pathogen interactions. However, how pathogen-derived EVs act on host cells is still poorly documented.
ano.nymous@ccsd.cnrs.fr.invalid (Julia Papail) 22 Sep 2023
https://hal.inrae.fr/hal-04215082
