First Insight into the Technological Features of Lactic Acid Bacteria Isolated from Algerian Fermented Wheat Lemzeiet

Fermented cereals are part of the main traditional diets of many people in Africa, usually obtained from artisanal production.The intensification of their manufacturing, responding to the consumers demand, requires a better control to ensure their sanitary, nutritional, and taste qualities, hence, the need of selecting accurate and safe starter cultures. In the present study, 48 lactic acid bacteria (LAB) strains, previously isolated from Algerian fermented wheat lemzeiet, were analyzed for different technological properties. 14 LAB strains, belonging to Pediococcus pentosaceus, Enterococcus faecium, Lactobacillus curvatus, Lactobacillus brevis, and Leuconostoc mesenteroides species, decreased rapidly the pH of the flour extract brothclose to 4 or below. 91% of strains showed extracellular protease activity, but only 12% were amylolytics. 18 LAB strains inhibited or postponed the growth of three fungal targets Rhodotorula mucilaginosa UBOCC-A-216004, Penicillium verrucosum UBOCC-A-109221, and Aspergillus flavus UBOCC-A-106028. The strains belonging to Lactobacillus spp., Leuconostoc fallax, L. mesenteroides, and Weissella paramesenteroides were the most antifungal ones. Multiplex PCR for biogenic amines’ production did not reveal any of the genes involved in the production of putrescine, histamine, and tyramine for 17 of the 48 strains. The obtained results provided several candidates for use as starter culture in the future production of lemzeiet

New insights about phenotypic heterogeneity within Propionibacterium freudenreichii argue against its division into subspecies

Propionibacterium freudenreichii is widely used in Swiss-type cheese manufacture, where it contributes to flavour and eye development. It is currently divided into two subspecies, according to the phenotype for lactose fermentation and nitrate reduction (lac+/nit- and lac-/nit+ for P. freudenreichii subsp. shermanii and subsp. freudenreichii, respectively). However, the existence of unclassifiable strains (lac+/nit+ and lac-/nit-) has also been reported. The aim of this study was to revisit the relevance of the subdivision of P. freudenreichii into subspecies, by confirming the existence of unclassifiable strains. Relevant conditions to test the ability of P. freudenreichii for lactose fermentation and nitrate reduction were first determined, by using 10 sequenced strains, in which the presence or absence of the lactose and nitrate genomic islands were known. We also determined whether the subdivision based on lac/nit phenotype was related to other phenotypic properties of interest in cheese manufacture, in this case, the production of aroma compounds, analysed by gas chromatography-mass spectrometry, for a total of 28 strains. The results showed that a too short incubation time can lead to false negative for lactose fermentation and nitrate reduction. They confirmed the existence of four lac/nit phenotypes instead of the two expected, thus leading to 13 unclassifiable strains out of the 28 characterized (7 lac+/nit+ and 6 lac-/nit-). The production of the 15 aroma compounds detected in all cultures varied more within a lac/nit phenotype (up to 20 times) than between them. Taken together, these results demonstrate that the division of P. freudenreichii into two subspecies does not appear to be relevant.Fil: de Freitas, Rosangela. Universidade Federal de Viçosa. Departamento de Tecnologia de Alimentos; Brasil. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Madec, Marie Noelle. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Chuat, Victoria. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Maillard, Marie Bernadette. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Abeijon Mukdsi, Maria Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Falentin, Hélène. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Carvalho, Antonio Fernandes de. Universidade Federal de Viçosa. Departamento de Tecnologia de Alimentos; BrasilFil: Valence, Florence. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; FranciaFil: Thierry, Anne. Institut National de la Recherche Agronomique; Francia. Science et Technologie du Lait et de l; Franci

Caractérisation et conservation de la diversité bactérienne d’un lait fermenté traditionnel breton, le Gwell en lien avec la préservation d’une race locale de vache, la Bretonne Pie Noir

Le Gwell est un lait fermenté traditionnel spécifique de la Bretagne. Il est obtenu à partir de lait de vaches de race Bretonne Pie Noir, inoculé avec une portion de la fabrication précédente (appelé ferment) sans aucun recours à des levains commerciaux. Les productions de Gwell partagent une texture ferme et onctueuse et un gout frais et acidulé, avec des caractéristiques organoleptiques propres à chaque producteur. Les producteurs sont malheureusement parfois confrontés à la perte de leur ferment et doivent alors avoir recours à la solidarité d’autres producteurs pour réacquérir un ferment opérationnel. Ces pertes de ferments sont un frein au développement de la production de Gwell et donc à la valorisation de lait issu de vaches Bretonne Pie Noir. Cette race emblématique de la Bretagne, caractérisée par une rusticité hors du commun et un lait très riche en matière grasse totalisait au milieu du 19ème siècle près de 900 000 têtes. La modernisation des pratiques agricoles alliée à une orientation productiviste forte a conduit à une quasi extinction de l’espèce, ce qui a conduit à initier en 1976 un programme de sauvegarde de l’espèce. Le nombre de vaches s’élève ainsi aujourd’hui à près de 2500 femelles. La transformation du lait en Gwell est, pour les éleveurs, un moyen de valoriser la qualité du lait de Bretonne Pie Noir en conservant sa valeur ajoutée. Les éleveurs qui transforment le lait en Gwell œuvrent ainsi à la sauvegarde de l’espèce Bretonne Pie Noir, mais aussi à la préservation de la diversité microbienne, du patrimoine et des savoir-faire paysans associés. La caractérisation de l’écosystème microbien du ferment Gwell, pour mieux maitriser sa conservation et sécuriser ainsi la production de Gwell, participe de ce fait au maintien de la race Bretonne Pie Noir. Dans ce contexte notre étude visait à caractériser l’écosystème microbien du Gwell pour sécuriser les souches à l’origine de la typicité du produit. Nous avons ainsi montré que toutes les productions de Gwell avaient une flore bactérienne dominante similaire, composée de deux sous-espèces de la bactérie lactique Lactococcus lactis (subsp. lactis et subsp. cremoris). En fonction des producteurs, le nombre de souches de chaque sous-espèce peut varier avec dans certain cas la présence de Streptococcus thermophilus. De plus, nous avons identifié et caractérisé des souches spécifiques à chaque producteur et montré une forte résilience de l’écosystème pouvant expliquer en partie les différences organoleptiques observées entre les Gwell de différents producteurs

Characterizing, modelling and understanding the climate variability of the deep water formation in the North-Western Mediterranean Sea

Observing, modelling and understanding the climate-scale variability of the deep water formation (DWF) in the North-Western Mediterranean Sea remains today very challenging. In this study, we first characterize the interannual variability of this phenomenon by a thorough reanalysis of observations in order to establish reference time series. These quantitative indicators include 31 observed years for the yearly maximum mixed layer depth over the period 1980–2013 and a detailed multi-indicator description of the period 2007–2013. Then a 1980–2013 hindcast simulation is performed with a fully-coupled regional climate system model including the high-resolution representation of the regional atmosphere, ocean, land-surface and rivers. The simulation reproduces quantitatively well the mean behaviour and the large interannual variability of the DWF phenomenon. The model shows convection deeper than 1000 m in 2/3 of the modelled winters, a mean DWF rate equal to 0.35 Sv with maximum values of 1.7 (resp. 1.6) Sv in 2013 (resp. 2005). Using the model results, the winter-integrated buoyancy loss over the Gulf of Lions is identified as the primary driving factor of the DWF interannual variability and explains, alone, around 50 % of its variance. It is itself explained by the occurrence of few stormy days during winter. At daily scale, the Atlantic ridge weather regime is identified as favourable to strong buoyancy losses and therefore DWF, whereas the positive phase of the North Atlantic oscillation is unfavourable. The driving role of the vertical stratification in autumn, a measure of the water column inhibition to mixing, has also been analyzed. Combining both driving factors allows to explain more than 70 % of the interannual variance of the phenomenon and in particular the occurrence of the five strongest convective years of the model (1981, 1999, 2005, 2009, 2013). The model simulates qualitatively well the trends in the deep waters (warming, saltening, increase in the dense water volume, increase in the bottom water density) despite an underestimation of the salinity and density trends. These deep trends come from a heat and salt accumulation during the 1980s and the 1990s in the surface and intermediate layers of the Gulf of Lions before being transferred stepwise towards the deep layers when very convective years occur in 1999 and later. The salinity increase in the near Atlantic Ocean surface layers seems to be the external forcing that finally leads to these deep trends. In the future, our results may allow to better understand the behaviour of the DWF phenomenon in Mediterranean Sea simulations in hindcast, forecast, reanalysis or future climate change scenario modes. The robustness of the obtained results must be however confirmed in multi-model studies

Des chercheurs entre science et industrie.

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Altering renneting pH changes microstructure, cell distribution and lysis of Lactococcus lactis AM2 in cheese made from ultrafiltered milk

The objective of this study was to investigate the lysis of a highly autolytic strain of Lactococcus lactis ssp. cremoris AM2 in a model cheese made from concentrated ultrafiltered milk. From the same initial ultrafiltered retentate inoculated with L. lactis AM2, 5 cheeses were made by the addition of rennet at different pH values (6.6, 6.2, 5.8, 5.4, and 5.2). Lysis was monitored by measurement of the release of lactate dehydrogenase, an intracellular marker enzyme, and by immunodetection of intracellular proteins with species-specific antibodies. Confocal scanning laser microscopy (CSLM) was used to investigate the cheese microstructure by staining for protein and fat. Dual staining with a bacterial viability kit with CSLM was performed to reveal the integrity and localization of the bacterial cells. Levels of soluble calcium significantly increased when the pH at which the rennet was added decreased. In cheese renneted at pH 6.6, CSLM revealed an open porous structure containing a dense protein network with fat globules of different sizes distributed in the aqueous phase. In cheese renneted at pH 5.2, the protein network was homogeneous, with a less dense protein network, and an even distribution of fat globules. On d 1, bacterial cells were organized into colonies in cheese renneted at pH 6.6, whereas in ch

Porosity of [i]Lactococcus lactis[/i] subsp. [i]lactis[/i] LD61 colonies immobilised in model cheese

During cheese ripening, micro-organisms grow as immobilised colonies, metabolising substrates present in the matrix which generate products triggered by enzymatic reactions. Local limitation rates of diffusion, either in the matrix or within the bacterial colonies, can be responsible for modulation in the metabolic and enzymatic activities of micro-organisms during ripening. How bacterial colonies immobilised in cheese are porous to these diffusing solutes has never been explored. The objective of this study was to determine if fluorescent dextrans of different sizes (4.4, 70 and 155 kDa) are able to penetrate through colonies of Lactococcus lactis LD61 immobilised in solid media, either agar or model cheese. Confocal microscopic observations showed that lactococcus colonies immobilised in these two media were porous to dextrans from 4 kDa to 155 kDa. However, the rate of diffusion of the solutes was faster inside the colonies immobilised in ultrafiltered-cheese than in agar when large dextrans were considered (≥70 kDa). The colonial shape of the lactococcus strain was also shown to be lenticular in agar and spherical in the model cheese, indicating that the physical pressure exerted on the colony by the surrounding casein network was probably isotropous in the UF-cheese but not in agar. In both cases, the fact that lactococcus colonies immobilised in solid media are porous to large dextran solutes suggests that substrates or enzymes are likely also to be able to migrate inside the colonies during cheese ripening

Altering renneting pH changes microstructure, cell distribution and lysis of Lactococcus lactis AM2 in cheese made from ultrafiltered milk

The objective of this study was to investigate the lysis of a highly autolytic strain of Lactococcus lactis ssp. cremoris AM2 in a model cheese made from concentrated ultrafiltered milk. From the same initial ultrafiltered retentate inoculated with L. lactis AM2, 5 cheeses were made by the addition of rennet at different pH values (6.6, 6.2, 5.8, 5.4, and 5.2). Lysis was monitored by measurement of the release of lactate dehydrogenase, an intracellular marker enzyme, and by immunodetection of intracellular proteins with species-specific antibodies. Confocal scanning laser microscopy (CSLM) was used to investigate the cheese microstructure by staining for protein and fat. Dual staining with a bacterial viability kit with CSLM was performed to reveal the integrity and localization of the bacterial cells. Levels of soluble calcium significantly increased when the pH at which the rennet was added decreased. In cheese renneted at pH 6.6, CSLM revealed an open porous structure containing a dense protein network with fat globules of different sizes distributed in the aqueous phase. In cheese renneted at pH 5.2, the protein network was homogeneous, with a less dense protein network, and an even distribution of fat globules. On d 1, bacterial cells were organized into colonies in cheese renneted at pH 6.6, whereas in ch

Comparison of amplified ribosomal DNA restriction analysis, peptidoglycan hydrolase and biochemical profiles for rapid dairy propionibacteria species identification

Species of dairy propionibacteria are used as cheese-ripening cultures as well as probiotics. However, no rapid identification methods are currently available. With this in mind, the present study compared three methods, (i) carbohydrate fermentation, (ii) ARDRA (amplified ribosomal DNA restriction analysis) and (iii) peptidoglycan hydrolase (PGH) activity profiles to improve the identification of Propionibacterium thoenii, Propionibacterium jensenii, Propionibacterium acidipropionici and Propionibacterium microaerophilum. The species Propionibacterium freudenreichii and Propionibacterium cyclohexanicum have previously been shown to be easily distinguishable from the other species. Principal component analysis of the carbohydrate fermentation profiles of 113 P. thoenii, P. jensenii, P. acidipropionici and P. microaerophilum strains correctly classified 85% of the strains based on the fermentation of seven carbohydrates. Regarding PGH profiles, optimized conditions of PGH-renaturing SDS–PAGE were applied to 34 of the strains. The PGH profiles of P. acidipropionici and P. microaerophilum were indistinguishable from one another, but were easily distinguished from P. jensenii and P. thoenii. However, four strains exhibited atypical profiles. Hence, in general, the PGH profiles were shown to be conserved within a species, with some exceptions. Four endonucleases were tested for ARDRA and the four species differentiated by combining the profiles obtained with MspI and HaeIII. P. freudenreichii and P. cyclohexanicum profiles were also performed but showed wide differences. Consequently, ARDRA was shown to be the most appropriate method for rapidly distinguishing strains of propionibacteria. Carbohydrate fermentation and peptidoglycan hydrolase activity profiles are useful as complementary identification tools, since about 15% of the 34 strains tested showed atypical profiles

Apo α-lactalbumin and lysozyme are colocalized in their subsequently formed spherical supramolecular assembly

We have reported previously that the calcium-depleted form of bovine a-lactalbumin (apo a-LA) interacts with hen egg-white lysozyme (LYS) to form spherical supramolecular structures. These supramolecular structures contain an equimolar ratio of the two proteins. We further explore here the organization of these structures. The spherical morphology and size of the assembled LYS⁄ apo a-LA supramolecular structures were demonstrated using confocal scanning laser microscopy and scanning electron microscopy. From confocal scanning laser microscopy experiments with labelled proteins, it was found that LYS and apo a-LA were perfectly colocalized and homogeneously distributed throughout the entire three-dimensional structure of the microspheres formed. The spatial colocalization of the two proteins was also confirmed by the occurrence of a fluorescence resonance energy transfer phenomenon between labelled apo a-LA and labelled LYS. Polarized light microscopy analysis revealed that the microspheres formed differ from spherulites, a higher order semicrystalline structure. As the molecular mechanism initiating the formation of these microspheres is still unknown, we discuss the potential involvement of a LYS⁄ apo a-LA heterodimer as a starting block for such a supramolecular assembly