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Nivoliez A.,Departement Recherche et Developpement Probionov | Nivoliez A.,CNRS Microorganisms Laboratory: Genome and Environment | Camares O.,Clermont University | Paquet-Gachinat M.,Clermont University | And 3 more authors.
Journal of Biotechnology | Year: 2012

Probiotics are administered as complex manufactured products and yet most studies on probiotic bacterial strains have been performed with native culture strains. Little is known about the influence of industrial processes on the properties of the microorganisms. In this study, we comparatively assessed the characteristics of the probiotic bacterial strain Lactobacillus rhamnosus (Lcr35®) together with four of its commercial formulations, including three intestinal formulas (BACILOR with Lcr Restituo® packet and capsule and FLOREA Lcr Lenio®) and one vaginal formula (GYNOPHILUS Lcr Regenerans®). Lcr35® grown from the intestinal formulas displayed increased resistance to acidic pH and bile stress, especially FLOREA (Lcr Lenio®), which showed a 4.5log higher number of viable bacteria compared to the results obtained with the control native Lcr35® strain. Adhesion to intestinal cells was significantly higher with Lcr Restituo® packet and Lcr Restituo® capsule vs Lcr35®. Bacteria from the vaginal formulation GYNOPHILUS had increased ability to metabolize glycogen thereby increasing lactic acid production. In vitro growth inhibition of the pathogen Candida albicans was significantly higher with bacteria from the vaginal formulation (4.5log difference) and in the presence of vaginal epithelial cells than with the native strain. Our results show that the manufacturing process influences strain properties and should therefore be adapted according to the strain and the therapeutic indication. © 2012 Elsevier B.V.

Muller C.,Departement Recherche et Developpement Probionov | Muller C.,University Paris - Sud | Mazel V.,University Paris - Sud | Dausset C.,Departement Recherche et Developpement Probionov | And 4 more authors.
European Journal of Pharmaceutics and Biopharmaceutics | Year: 2014

The beneficial effects of probiotic bacteria on human health are now widely acknowledged, and this has prompted growing interest in research and development in the pharmaceutical field. However, to be viable when they reach their target, the bacteria must be able to survive during the manufacturing process and the biological pathway. Tablet form best meets the requirements for protecting acid labile drugs, but the tableting process could be an additional stress for the bacteria. This study evaluated the initial effect of compression pressure on the Lcr35® strain in a vaginal (Lcr regenerans®) and an intestinal (Lcr restituo®) formulation. A stability study was also performed on the tablets and revealed a beneficial effect of this form. The obtained destruction rates (k) demonstrated that the bacterial stability was greater in tablets than in powders (kpowders > ktablets). A new mathematical model was developed combining compression and temperature parameters to predict the bacterial viability at any pressure and time. Moreover, the genetic profile of Lcr35® (Rep-PCR, microarrays), its resistance to acidity and its ability to inhibit Candida albicans growth, after compression, were determined to evaluate the target product profile (TPP) in a Quality by Design (QbD) approach. The Rep-PCR analysis validated the strain identity and the microarrays demonstrated the genetic stability of Lcr35® strain after compaction. Additionally, ability to inhibit the C. albicans growth was maintained and the resistance to gastric conditions of Lcr35® was even improved by tableting. As a dosage form, tablets containing probiotic can guarantee that an adequate amount of bacteria reaches the therapeutic target (intestinal or vaginal) and that the product remains stable until the time of consumption. © 2014 Elsevier B.V. All rights reserved.

Nivoliez A.,Departement Recherche et Developpement Probionov | Nivoliez A.,CNRS Microorganisms Laboratory: Genome and Environment | Veisseire P.,Clermont University | Alaterre E.,Departement Recherche et Developpement Probionov | And 7 more authors.
Applied Microbiology and Biotechnology | Year: 2014

The influence of the industrial process on the properties of probiotics, administered as complex manufactured products, has been poorly investigated. In the present study, we comparatively assessed the cell wall characteristics of the probiotic strain Lactobacillus rhamnosus Lcr35® together with three of its commercial formulations with intestinal applications. Putative secreted and transmembrane-protein-encoding genes were initially searched in silico in the genome of L. rhamnosus Lcr35®. A total of 369 candidate genes were identified which expressions were followed using a custom Lactobacillus DNA chip. Among them, 60 or 67 genes had their expression either upregulated or downregulated in the Lcr Restituo® packet or capsule formulations, compared to the native Lcr35® strain. Moreover, our data showed that the probiotic formulations (Lcr Lenio®, Lcr restituo® capsule and packet) showed a better capacity to adhere to intestinal epithelial Caco-2 cells than the native Lcr35® strain. Microbial (MATS) tests showed that the probiotic was an electron donor and that they were more hydrophilic than the native strain. The enhanced adhesion capacity of the active pharmaceutical ingredients (APIs) to epithelial Caco-2 cells and their antipathogen effect could be due to this greater surface hydrophilic character. These findings suggest that the manufacturing process influences the protein composition and the chemical properties of the cell wall. It is therefore likely that the antipathogen effect of the formulation is modulated by the industrial process. Screening of the manufactured products’ properties would therefore represent an essential step in evaluating the effects of probiotic strains. © 2014, Springer-Verlag Berlin Heidelberg.

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