Microbiological, physicochemical and sensory changes throughout ripening of an experimental soft smear-ripened cheese in relation to salt concentrations.

To evaluate the sodium chloride content effect on microbiological, biochemical, physicochemical and sensorial characteristics, Munster cheeses were prepared from pasteurized milk seeded with 3 yeasts (Kluyveromyces marxianus, Debaryomyces hansenii, Geotrichum candidum) and 5 ripening bacteria (Arthrobacter arilaitensis, Brevibacterium aurantiacum, Corynebacterium casei, Hafnia alvei, and Staphylococcus equorum). Experiments were performed under 1.0%, 1.7% and 2.4% NaCl levels in cheese in triplicate. Ripening (d2 - d27) was carried under 12°C and 96% RH. These kinetics were both reproducible and repeatable at 99% confidence level. For each microbial, biochemical and physicochemical parameter, 2 kinetic descriptors (maximal or minimal rate and its occurrence time) were defined. On d2 the physicochemical variables (water activity, dry matter, water content) were strongly dependent on the salting level. From d2 to d27 K. lactis was insensitive to salt while D. hansenii was stimulated. G. candidum growth appeared very sensitive to salt in cheese: at 1.0% NaCl G. candidum exhibited overgrowth, negatively impacting rind appearance, underrind consistency and thickness and off-flavor flaws. Salt concentration of 2.4% induced death of G. candidum. Four bacteria (A. arilaitensis, B. aurantiacum, C. casei, and H. alvei) were moderately sensitive to salt while S. equorum was insensitive to it. Salt level in cheese had a significant effect on carbon substrate consumption rates. Lactate consumption rate in 1.0% salted cheeses was approximately twice higher than under 2.4% NaCl. Data analysis of microorganism, biochemical, and physicochemical kinetics and sensory analysis showed that the best salt level in Munster-type cheeses to achieve an optimum balance between cheese characteristics, sensory qualities and marketability was 1.7% NaCl.

A rapid method for the assessment of the vitality of microorganisms using flow cytometry.

Cultivability, viability, and vitality make it possible to characterize the behavior of a cellular population. Vitality was assessed using the kinetic parameters of specific metabolisms depending on whether the strains were used, for example, for the acidification of lactic acid bacteria or for CO2 production in fermenting yeasts. However, these methods are time-consuming. We developed a cytometric descriptor based on the energy-dependent extrusion of carboxyfluorescein from cells, subsequent to carboxyfluorescein diacetate staining, and compared it to the measurements of metabolic activities of various bacteria and yeasts. For all of the microorganisms tested, the cytometric descriptor ΔFI15 was well correlated with the results of the metabolic measurements and, moreover, has the advantage of being easier and faster to use than metabolic methods. It can be very useful for evaluating the vitality of the starters before inoculation in industrial processes.

Food-dependent set-up of the DiDGI® dynamic in vitro system: Correlation with the porcine model for protein digestion of soya-based food.

The present study compared in vivo protein digestion in a miniature pig model with the dynamic in vitro system DiDGI®, using three digestive compartments (stomach, duodenum, and jejunum + ileum). Two soya-based meals-commercial soya milk and tofu-were studied, each with the same macronutrient content but different macrostructures. Our aim was to first deduce from the in vivo experiments in pigs key digestive parameters such as gastric pH, stomach emptying kinetics, and intestinal transit time, in order to design a relevant set-up for the dynamic in vitro system. Then, we compared digestive samples collected at fixed sampling times from both in vivo and in vitro models regarding different values related to proteolysis. We observed similar evolutions of gastric peptide distribution and duodenal proteolysis between models. Overall, apparent ileal digestibility of nitrogen was similar in vitro and in vivo and the differences between the two meals were conserved between models.

Physicochemical and sensory evolutions of the lactic goat cheese Picodon in relation to temperature and relative humidity used throughout ripening.

To produce a wide variety of cheeses, it is necessary to control the ripening process. To do that, artisanal goat cheeses were ripened to evaluate the effects of temperature (10 and 14°C) and relative humidity (RH; 88 and 98%) on (1) 16 physicochemical characteristics throughout ripening and (2) 19 sensory characteristics at the end of ripening (d 12). Whatever the ripening time, the physicochemical characteristics were strongly dependent on the daily productions, which affected the sensory perception of the cheeses. Both physicochemical and sensory characteristics were strongly reliant on RH, whereas only a few of the characteristics were influenced by temperature changes. On d 12, whatever the ripening temperature, an RH increase from 88% to 98% modified many cheese characteristics (core pH, lactate consumption, underrind thickening, dry matter content, and hardness). As a result of these physicochemical properties, changes in perception were observed: the cheeses ripened under 88% RH were dry and hard compared with those ripened under 98% RH. An RH of 98% led to an acceleration of the ripening process, inducing a slightly ammonia and milky flavor and a sticky and creamy texture in the mouth. However, cheeses ripened under 14°C and 98% RH were also indicative of overripened cheeses: a temperature of 14°C induced an acceleration of the ripening process due to physicochemical modifications compared with a temperature of 10°C. Nevertheless, when the cheeses on d 0 were still very humid and soft, those ripened under 98% RH collapsed and were overripened with a liquid underrind. This study provides a means for achieving a better and more rational control of the ripening process in artisanal lactic goat cheeses.

Effect of dairy matrices on the survival of Streptococcus thermophilus, Brevibacterium aurantiacum and Hafnia alvei during digestion.

This study evaluated the ability of dairy matrices, different in composition (with and without fat) and structure (liquid and gel), to enhance microorganisms survival through digestion. The viability of three dairy microorganisms Streptococcus thermophilus, Brevibacterium aurantiacum and Hafnia alvei was measured during in vitro and in vivo digestion. S. thermophilus was highly sensitive to gastric stress, and was not found in the duodenal compartment. B. auranticum was moderately sensitive to gastric stress but resistant to duodenal stress. H. alvei was highly resistant to both stresses. LIVE/DEAD confocal microscopy's images, probed the effect of low pH on microorganisms survival. However, in vivo analyses (16S rRNA gene metabarcoding) failed to confirm in vitro observations since tested microorganisms were not detected. Despite of the different evolutions during digestion on buffer capacity, lipolysis, and rheological characteristics, we did not observe any protective effect of the dairy matrices on microorganisms survival.

Survival of cheese-ripening microorganisms in a dynamic simulator of the gastrointestinal tract.

A mixture of nine microorganisms (six bacteria and three yeasts) from the microflora of surface-ripened cheeses were subjected to in vitro digestive stress in a three-compartment "dynamic gastrointestinal digester" (DIDGI). We studied the microorganisms (i) grown separately in culture medium only (ii) grown separately in culture medium and then mixed, (iii) grown separately in culture medium and then included in a rennet gel and (iv) grown together in smear-ripened cheese. The yeasts Geotrichum candidum, Kluyveromyces lactis and Debaryomyces hansenii, were strongly resistant to the whole DIDGI process (with a drop in viable cell counts of less than <1 log CFU mL(-1)) and there were no significant differences between lab cultures and cheese-grown cultures. Ripening bacteria such as Hafnia alvei survived gastric stress less well when grown in cheese (with no viable cells after 90 min of exposure of the cheese matrix, compared with 6 CFU mL(-1) in lab cultures). The ability of Corynebacterium casei and Staphylococcus equorum to withstand digestive stress was similar for cheese and pure culture conditions. When grow in a cheese matrix, Brevibacterium aurantiacum and Arthrobacter arilaitensis were clearly more sensitive to the overall digestive process than when grown in pure cultures. Lactococcus lactis displayed poorer survival in gastric and duodenal compartments when it had been grown in cheese. In vivo experiments in BALB/c mice agreed with the DIDGI experiments and confirmed the latter's reliability.

A Decision Support System Coupling Fuzzy Logic and Probabilistic Graphical Approaches for the Agri-Food Industry: Prediction of Grape Berry Maturity.

Agri-food is one of the most important sectors of the industry and a major contributor to the global warming potential in Europe. Sustainability issues pose a huge challenge for this sector. In this context, a big issue is to be able to predict the multiscale dynamics of those systems using computing science. A robust predictive mathematical tool is implemented for this sector and applied to the wine industry being easily able to be generalized to other applications. Grape berry maturation relies on complex and coupled physicochemical and biochemical reactions which are climate dependent. Moreover one experiment represents one year and the climate variability could not be covered exclusively by the experiments. Consequently, harvest mostly relies on expert predictions. A big challenge for the wine industry is nevertheless to be able to anticipate the reactions for sustainability purposes. We propose to implement a decision support system so called FGRAPEDBN able to (1) capitalize the heterogeneous fragmented knowledge available including data and expertise and (2) predict the sugar (resp. the acidity) concentrations with a relevant RMSE of 7 g/l (resp. 0.44 g/l and 0.11 g/kg). FGRAPEDBN is based on a coupling between a probabilistic graphical approach and a fuzzy expert system.

In vitro characterization of the digestive stress response and immunomodulatory properties of microorganisms isolated from smear-ripened cheese.

Thirty-six microorganisms (twenty-one bacteria, twelve yeasts and three fungi) were isolated from surface-ripened cheeses and subjected to in vitro digestive stress. The approach mimicked gastric and/or duodenal digestion. Lactobacillus rhamnosus GG, Escherichia coli Nissle 1917 and Saccharomyces boulardii were used as reference strains. We studied the microorganisms grown separately in culture medium and then included (or not) in a rennet gel. The microorganisms' immunomodulatory abilities were also assessed by profiling cytokine induction in human peripheral blood mononuclear cells (PBMCs). The loss of viability was less than 1 log CFU/mL for yeasts under all conditions. In contrast, Gram-negative bacteria survived gastric and/or duodenal stress well but most of the Gram-positive bacteria were more sensitive (especially to gastric stress). Inclusion of sensitive Gram-positive bacteria in rennet gel dramatically improved gastric survival, when compared with a non-included cultured (with a 4 log CFU/mL change in survival). However, the rennet gel did not protect the bacteria against duodenal stress. The PBMC cytokine assay tests showed that the response to yeasts was usually anti-inflammatory, whereas the response to bacteria varied from one strain to another.

Validation of a new in vitro dynamic system to simulate infant digestion.

Understanding the mechanisms of infant formula disintegration in the infant gastrointestinal tract is a key step for developing new formulas with health benefits for the neonate. For ethical reasons, the access to in vivo data obtained on infants is limited. The use of animal models can be an alternative but these experiments are labour intensive, expensive and results obtained show high inter-individual variability, making their interpretation difficult. The aim of this work was to develop a simple in vitro dynamic gastrointestinal digestion system, for studying infant formula digestion, and to validate it by comparing the kinetics of proteolysis obtained in vitro with in vivo data collected from piglets. Results showed a good correlation between in vitro and in vivo data and confirmed the rapid hydrolysis of caseins in gastric conditions, whereas whey proteins appeared more resistant to digestion.

Dynamics of Penicillium camemberti growth quantified by real-time PCR on Camembert-type cheeses under different conditions of temperature and relative humidity.

Penicillium camemberti plays a major role in the flavor and appearance of Camembert-type cheeses. However, little is known about its mycelium growth kinetics during ripening. We monitored the growth of P. camemberti mycelium in Camembert-type cheeses using real-time PCR in 4 ripening runs, performed at 2 temperatures (8 and 16°C) and 2 relative humidities (88 and 98%). These findings were compared with P. camemberti quantification by spore concentration. During the first phase, the mycelium grew but no spores were produced, regardless of the ripening conditions. During the second phase, which began when lactose was depleted, the concentration of spores increased, especially in the cheeses ripened at 16°C. Sporulation was associated with a large decrease in the mycelial concentration in the cheeses ripened at 16°C and 98% relative humidity. It was hypothesized that lactose is the main energy source for the growth of P. camemberti mycelium at the beginning of ripening and that its depletion would trigger stress, resulting in sporulation.

Discrimination of Cognacs and other distilled drinks by mid-infrared spectropscopy.

Mid-infrared spectroscopy was applied to the analysis and discrimination of Cognacs and other distilled drinks (Armagnacs, whiskies, brandies, bourbons, rums, and counterfeit products). Strong correlations were found between dry extract spectra, polyphenolic dry extract spectra, and the total polyphenol concentration of samples, notably of Cognacs. Principal component analysis applied to spectral data made it possible to emphasize the importance of dry extract data when a distinction is made between Cognacs and Armagnacs, whiskies, bourbons, and rums, and of polyphenol concentration when Cognacs, brandies, and counterfeit products are separated. Ninety-six percent of samples in the test set were correctly assigned to Cognacs and non-Cognacs by partial least-squares discriminant analysis.