Mesure de l’acidité des zéolites par adsorption d’indicateurs colorés

La détermination du nombre et de la force des sites acides des zéolites est réalisée par une méthode de titrage indirecte établie à partir du déplacement par la pyridine d’une fraction d’indicateur coloré préalablement adsorbé à la surface du solide. Pour que les résultats soient rigoureux, les indicateurs utilisés sont des anilines primaires et appartiennent à la même fonction d’acidité H’o de Hammet ; les mesures sont effectuées par spectrophotométrie UV. Nous mettons en évidence une sigmoide de protonation analogue à celle obtenue en milieu homogène dans les titrages acide-base classiques

Sélection et intégration d'une souche probiotique fonctionnelle dans une matrice sèche

Les probiotiques sont des micro-organismes vivants, capables d interagir avec le microbiote et les cellules de l organisme hôte. Par leurs présences ou leurs métabolismes, ils contribuent notamment à la régulation du système immunitaire. D un point de vue technologique, le développement d un complément alimentaire contenant des probiotiques nécessite la maitrise des étapes de sélection et de production d une bactérie fonctionnelle sous forme sèche et revivifiable. Au cours de ce travail, nous avons développé des tests de criblage en cytométrie en flux, permettant de déterminer l aptitude d une bactérie à être séchée et l origine de sa sensibilité. Pour mettre en place ces tests, nous nous sommes intéressés aux mécanismes mis en jeu lors du procédé de séchage. Ainsi, lors de la déshydratation, les micro-organismes vont subir simultanément un stress osmotique et un stress oxydant. Les résultats montrent un fort impact du séchage sur la viabilité et le potentiel immuno-modulateur avec une diminution de la composante pro-inflammatoire (IL-12). Pour protéger les bactéries, trois stratégies d optimisation ont été développées : le séchage par formation de mousse, l encapsulation dans une matrice alginate-protéines de pois, et le co-séchage d une bactérie fragile avec une bactérie résistante. Les connaissances acquises au cours de cette étude ont permis d optimiser la production industrielle de la souche finale du projet. Afin de répondre au nouveau cadre réglementaire européen, une étude clinique en double aveugle contre placébo est actuellement en cours (2011-2013)Probiotics are live microorganisms that can act on the immune system at specific sites in the gastro-intestinal tract. From a technological point of view, the development of a dietary supplement containing viable and functional probiotics requires the control of the selection and production steps. In a first time, we have developed specific and sensitive screening tests to determine the ability of bacteria to be dried and the origin of its sensitivity. To develop these tests, we investigated the mechanisms involved during drying process. Thus, microorganisms simultaneously undergo osmotic stress and oxidative stress. Results show a strong impact of drying on the survival rate and immunomodulatory potential with a decrease of proinflammatory cytokines (IL-12). To protect probiotic bacteria, three optimization strategies have been developed: drying by foam formation, encapsulation of probiotic bacteria using a pea-protein alginate matrix and drying of a sensitive probiotic strain with a resistant bacterium. Knowledge obtained during this study was used to optimize the industrial process of the final strain of the project. A randomized double blinded (RDB) trial is in progress (2011-2013) to test the bacteria in humansDIJON-BU Doc.électronique (212319901) / SudocSudocFranceF

Industrial production of dried yeast: plasma membrane as a survival indicator of air drying process

International audiencePreservation of microorganisms by desiccationis a major industrial interest.However, study of cell survival mechanismsthat occur during desiccation is complex. In this work, the impact of the magnitude and the kinetics of dehydration on yeastsurvival wereevaluatedin either hyperosmotic liquid medium or a gaseous environment. Asame lethal magnitude of dehydration and a same lethal kinetic effect were found. As previously shown for osmotic stress, this work demonstrate that yeast survival after drying was also related to plasma membrane disorganization, suggesting a similar passive rearrangement of the membrane components.Hyperosmotic treatment in liquid medium represents an opportune environment to optimize industrial drying processes

Air-drying kinetics affect yeast membrane organization and survival.

International audienceThe plasma membrane (PM) is a key structure for the survival of cells during dehydration. In this study, we focused on the concomitant changes in survival and in the lateral organization of the PM in yeast strains during desiccation, a natural or technological environmental perturbation that involves transition from a liquid to a solid medium. To evaluate the role of the PM in survival during air-drying, a wild-type yeast strain and an osmotically fragile mutant (erg6Δ) were used. The lateral organization of the PM (microdomain distribution) was observed using a fluorescent marker related to a specific green fluorescent protein-labeled membrane protein (Sur7-GFP) after progressive or rapid desiccation. We also evaluated yeast behavior during a model dehydration experiment performed in liquid medium (osmotic stress). For both strains, we observed similar behavior after osmotic and desiccation stresses. In particular, the same lethal magnitude of dehydration and the same lethal kinetic effect were found for both dehydration methods. Thus, yeast survival after progressive air-drying was related to PM reorganization, suggesting the positive contribution of passive lateral rearrangements of the membrane components. This study also showed that the use of glycerol solutions is an efficient means to simulate air-drying desiccation

Ergosterol biosynthesis: a fungal pathway for life on land?

International audienceSterols, essential lipids of most eukaryotic cells, ensure important structural and signaling functions. The selection pressure that has led to different dominant sterols in the three eukaryotic kingdoms remains unknown. Here, we investigated the influence of the progression in the different steps of the ergosterol biosynthetic pathway (EBP) on the yeast resistance to transitions from aqueous to aerial media, typical perturbations of the higher fungi habitats. Five mutants of the EBP (ergΔ), accumulating different sterol intermediates in the EBP, and the wild-type (WT) strain were exposed to drying under atmospheric air or nitrogen and wetting. Results show that the progression in the EBP parallels an increase in the yeast resistance to air-drying with a maximal survival rate for the WT strain. When drying/wetting was performed under nitrogen, yeast survival was higher, particularly for the earlier mutants of the EBP. Thus, ergosterol, through its protective role against mechanical and oxidative stress, might have been selected by the pressure induced by drying/wetting cycles occurring in the fungi habitats. These results support the Bloch hypothesis, which postulates that the properties of sterols are gradually optimized for function along the biosynthetic pathway and provide a response to the enduring question "why ergosterol in fungi?"

Impact of probiotics on risk factors for cardiovascular diseases. A review.

International audienceProbiotic microorganisms have historically been used to rebalance disturbed intestinal microbiota and to diminish gastrointestinal disorders, such as diarrhea or inflammatory bowel diseases (e.g., Crohn's disease and ulcerative colitis). Recent studies explore the potential for expanded uses of probiotics on medical disorders that increase the risk of developing cardiovascular diseases and diabetes, such as obesity, hypercholesterolemia, arterial hypertension, and metabolic disturbances such as hyperhomocysteinemia and oxidative stress. This review aims at summarizing the proposed molecular and cellular mechanisms involved in probiotic-host interactions and to identify the nature of the resulting beneficial effects. Specific probiotic strains can act by modulating immune response, by producing particular molecules or releasing biopeptides, and by modulating nervous system activity. To date, the majority of studies have been conducted in animal models. New investigations on the related mechanisms in humans need to be carried out to better enable targeted and effective use of the broad variety of probiotic strains

A way to follow the viability of encapsulated Bifidobacterium bifidum subjected to a freeze-drying process in order to target the colon: Interest of flow cytometry

International audienceThe aim of this work was to apply flow cytometry in order to assess and compare the viability of freeze-dried entrapped bacteria with an usual technique by quantification by plate count techniques. It also aimed at studying the effect of various cryoprotectants on the viability of an entrapped Bifidobacterium bifidum subjected to freeze-drying to check their ability to be delivered all along the gastro-intestinal tract. The alginate–pectinate beads were chosen as the encapsulation matrix added with different protectants. The beads were characterized by scanning electron microscopy and the viability was checked by both methods. The best combination to improve viability of entrapped bacteria subjected to freeze-drying is made of glycerol 20% (one cryoprotectant) and sodium ascorbate 10% (one anti-oxidative compound). This study also demonstrates that flow cytometry allows assessment of entrapped bacteria viability. Indeed we showed that viability evaluated by plate method is correlated to that obtained by flow cytometry. So, flow cytometry is a rapid method to determine cell viability after encapsulation and freeze-drying. Finally, these beads seem to be a promising probiotic delivery system to target the colon

Drying process strongly affects probiotics viability and functionalities

International audienceProbiotic formulations are widely used and are proposed to have a variety of beneficial effects, depending on the probiotic strains present in the product. The impact of drying processes on the viability of probiotics is well documented. However, the impact of these processes on probiotics functionality remains unclear. In this work, we investigated variations in seven different bacterial markers after various desiccation processes. Markers were composed of four different viability evaluation (combining two growth abilities and two cytometric measurements) and in three in vitro functionalities: stimulation of IL-10 and IL-12 production by PBMCs (immunomodulation) and bacterial adhesion to hexadecane. We measured the impact of three drying processes (air-drying, freeze-drying and spray-drying), without the use of protective agents, on three types of probiotic bacteria: Bifidobacterium bifidum, Lactobacillus plantarum and Lactobacillus zeae. Our results show that the bacteria respond differently to the three different drying processes, in terms of viability and functionality. Drying methods produce important variations in bacterial immunomodulation and hydrophobicity, which are correlated. We also show that adherence can be stimulated (air-drying) or inhibited (spray-drying) by drying processes. Results of a multivariate analysis show no direct correlation between bacterial survival and functionality, but do show a correlation between probiotic responses to desiccation–rewetting and the process used to dry the bacteria

Data

Data concerning the survival of the different yeast strains after drying treatments and yeast sterol analysi