In most ripened cheeses, bacteria are responsible for the ripening process. Immobilized in the cheese matrix, they grow as colonies. Therefore, their distribution as well as the distance between them are of major importance for ripening steps since metabolites diffuse within the cheese matrix. No data are available to date about the spatial distribution of bacterial colonies in cheese. This is the first study to model the distribution of bacterial colonies in a food-type matrix using nondestructive techniques. We compared (i) the mean theoretical three-dimensional (3D) distances between colonies calculated on the basis of inoculation levels and considering colony distribution to be random and (ii) experimental measurements using confocal microscopy photographs of fluorescent colonies of a Lactococcus lactis strain producing green fluorescent protein (GFP) inoculated, at different levels, into a model cheese made by ultrafiltration (UF). Enumerations showed that the final numbers of cells were identical whatever the inoculation level (10 4 to 10 7 CFU/g). Bacterial colonies were shown to be randomly distributed, fitting Poisson's model. The initial inoculation level strongly influenced the mean distances between colonies (from 25 m to 250 m) and also their mean diameters. The lower the inoculation level, the larger the colonies were and the further away from each other. Multiplying the inoculation level by 50 multiplied the interfacial area of exchange with the cheese matrix by 7 for the same cell biomass. We finally suggested that final cell numbers should be discussed together with inoculation levels to take into account the distribution and, consequently, the interfacial area of colonies, which can have a significant influence on the cheese-ripening process on a microscopic scale.During cheese making, regardless of the cheese type, bacteria are immobilized in the curd during the coagulation step. It is generally accepted that 90% of the bacteria present in the milk are retained, trapped in the curd, while only 10% are lost in the whey during draining (16). In cheeses made by ultrafiltration (UF), the draining step is absent, and 100% of the cells are then retained in the curd. In any case, after immobilization by coagulation, each inoculated bacterial cell is assumed to grow, generating a colony inside the curd. Colonies are then spread within the cheese curd, and they interact with the cheese matrix during ripening. Consequently, the ripening process must take place on a microscopic scale around colonies. Only studies showing microscopic examinations of bacterial colonies in cheese either by electronic microscopy (24) or, more recently, by confocal laser scanning microscopy (7, 19) have been reported.The ripening process (proteolysis, lipolysis, amino acid catabolism, and the production of organic acids, etc.) relies on the metabolic activities of bacterial colonies, leading to the formation of flavors and textures of cheese (11,25). So far, ripening has always been described with average processes on the che...
-In cheese technology, the mass transfer of small solutes, such as salt, moisture and metabolites during brining and ripening, is very important for the final quality of the cheese. This paper has the following objectives: (i) to review the data concerning the diffusion coefficients of solutes in different cheese types; (ii) to review the experimental methods available to model the mass transfer properties of small solutes in complex matrices such as cheese; and (iii) to consider some potential alternative approaches. Numerous studies have reported the transfer of salt in cheese during brining and ripening. Regardless of the type of cheese and its composition, the effective diffusion coefficients of salt have been reported to be between 1 and 5.3 × 10 −10 m 2 ·s −1 at 10-15°C. However, few papers have dealt with the mass transfer properties of other small solutes in cheese. Most of the reported effective diffusion coefficient values have been obtained by macroscopic and destructive concentration profile methods. More recently, some other promising techniques, such as nuclear magnetic resonance, magnetic resonance imaging or fluorescence recovery after photobleaching, are currently being developed to measure the mass transfer properties of solutes in heterogeneous media at microscopic scales. However, these methods are still difficult to apply to complex matrices such as cheese. Further research needs to focus on: (i) the development of nondestructive techniques to determine the mass transfer properties of small solutes at a microscopic level in complex matrices such as cheese; and (ii) the determination of the mass transfer properties of metabolites that are involved in enzymatic reactions during cheese ripening. cheese / mass transfer / diffusion / modelling / solute Article published by EDP SciencesRésumé -Détermination des coefficients de diffusion de petits solutés dans le fromage : une synthèse. En technologie fromagère, le transfert de petits solutés, tels que le sel, l'eau et les métabolites au cours du saumurage et de l'affinage, joue un rôle majeur sur la qualité finale du fromage. Cette revue bibliographique a pour objectifs principaux : (i) de faire le bilan des valeurs publiées des coefficients de diffusion de différents solutés dans les fromages ; (ii) de passer en revue les méthodes expérimentales disponibles pour déterminer les propriétés de transfert des petits solutés dans des milieux complexes comme le fromage ; (iii) de considérer les méthodes alternatives potentiellement applicables aux fromages. Dans la littérature, de nombreuses études ont été publiées au sujet du transfert de sel dans les fromages au cours du saumurage et de l'affinage. En fonction du type de fromage et de sa composition, les coefficients de diffusion effectifs du sel sont compris entre 1 et 5,3 × 10 −10 m 2 ·s −1 à des températures comprises entre 10 et 15°C. Très peu d'études concernant les propriétés de transfert d'autres petits solutés dans les fromages ont été publiées. La plupart des coefficients de diffusi...
The diffusion of small solutes in cheese is of key importance for most enzymatic reactions involved in the ripening process. However, only a limited amount of data is available on salt diffusion and practically none on peptide diffusion. Nisin, a bacteriocin peptide, migrated in model cheeses made from ultrafiltered (UF) retentate. A profile concentration device and an enzyme-linked immunosorbent assay (ELISA), specifically developed for nisin quantification in cheese, were used to model the apparent diffusion coefficients for nisin according to Fick's law. This average coefficient was 49.5 μm(2)/s in UF cheese (n = 2). When 10% gelatin was added to the retentate, this value decreased to 34.4 μm(2)/s (n = 2). The two cheeses differed in their macrostructure (rheology) and microstructure (confocal microscopy). This study provides the first apparent diffusion coefficients for a peptide in cheese and supports the hypothesis that composition and structure influence the diffusion of small solutes such as peptides.
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