RESUMEN
Perception of flavor is a dynamic process during which the concentration of aroma molecules at the olfactory epithelium varies with time as they are released progressively from the food in the mouth during consumption. The release kinetics depends on the food matrix itself but also on food oral processing, such as mastication behavior and food bolus formation with saliva, for which huge inter-individual variations exist due to physiological differences. Sensory methods such as time intensity (TI) or the more-recent methods temporal dominance of sensations (TDS) and temporal check-all-that-apply (TCATA) are used to account for the dynamic and time-related aspects of flavor perception. Direct injection mass spectrometry (DIMS) techniques that measure in real time aroma compounds directly in the nose (nosespace), aimed at obtaining data that reflect the pattern of aroma release in real time during food consumption and supposed to be representative of perception, have been developed over the last 25 years. Examples obtained with MS operated in chemical ionization mode at atmospheric or sub-atmospheric pressure (atmospheric pressure chemical ionization APCI or proton-transfer reaction PTR) are given, with emphases on studies conducted with simultaneous dynamic sensory evaluation. Inter-individual variations in terms of aroma release and their relevance for understanding flavor perception are discussed as well as the evidenced cross-modal interactions.
Asunto(s)
Líquidos Corporales , Odorantes , Presión Atmosférica , Cara , PercepciónRESUMEN
Since 1975, the Weurman Flavour Research Symposium has been held every three years in different European countries, and has been finally established as an international event that offers unique opportunities for distinguished scientists from academia and industry, from different disciplines, and from all over the world, to discuss trends and new paradigms in the field of flavour research [...].
Asunto(s)
Aromatizantes , Gusto , Europa (Continente)RESUMEN
The headspace volatile organic compound (VOC) fingerprints (volatilome) of French wine brandies were investigated by proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS). Protonated ethanol chemical ionization was used with dedicated experimental conditions that were previously validated for model wines. These included a reference vial containing a hydro-alcoholic solution with the same ethanol content (20% v/v) as the diluted sample spirits, which was used to establish steady-state ionization conditions. A low electric field strength to number density ratio E/N (85 Td) was used in the drift tube in order to limit the fragmentation of the protonated analytes. The obtained headspace fingerprints were used to investigate the origin of French brandies produced within a limited geographic production area. Brandies of two different vintages (one freshly distilled and one aged for 14 years in French oak barrels) were successfully classified according to their growth areas using unsupervised (principal component analysis, PCA) and supervised (partial least squares regression discriminant analysis, PLS-DA) multivariate analyses. The models obtained by PLS-DA allowed the identification of discriminant volatile compounds that were mainly characterised as key aroma compounds of wine brandies. The discrimination was supported by sensory evaluation conducted with free sorting tasks. The results showed that this ethanol ionization method was suitable for direct headspace analysis of brandies. They also demonstrated its ability to distinguish French brandies according to their growth areas, and this effect on brandy VOC composition was confirmed at a perceptive level.
Asunto(s)
Etanol/química , Espectrometría de Masas/métodos , Compuestos Orgánicos Volátiles/análisis , Vino/análisis , FranciaRESUMEN
Dark chocolate samples were previously classified into four sensory categories. The classification was modelled based on volatile compounds analyzed by direct introduction mass spectrometry of the chocolates' headspace. The purpose of the study was to identify the most discriminant odor-active compounds that should characterize the four sensory categories. To address the problem, a gas chromatography-olfactometry (GC-O) study was conducted by 12 assessors using a comparative detection frequency analysis (cDFA) approach on 12 exemplary samples. A nasal impact frequency (NIF) difference threshold combined with a statistical approach (Khi² test on k proportions) revealed 38 discriminative key odorants able to differentiate the samples and to characterize the sensory categories. A heatmap emphasized the 19 most discriminant key odorants, among which heterocyclic molecules (furanones, pyranones, lactones, one pyrrole, and one pyrazine) played a prominent role with secondary alcohols, acids, and esters. The initial sensory classes were retrieved using the discriminant key volatiles in a correspondence analysis (CA) and a hierarchical cluster analysis (HCA). Among the 38 discriminant key odorants, although previously identified in cocoa products, 21 were formally described for the first time as key aroma compounds of dark chocolate. Moreover, 13 key odorants were described for the first time in a cocoa product.
Asunto(s)
Chocolate/análisis , Odorantes/análisis , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/química , Cromatografía de Gases y Espectrometría de Masas , SensaciónRESUMEN
Olfactory mucosa (OM) can metabolise odorant volatile organic compounds through various enzymatic mechanisms to produce odorous or non-odorous metabolites. Preliminary ex vivo studies using headspace-gas chromatography (HS-GC) revealed the formation of metabolites when odorant molecules were injected in the headspace above a fresh explant of rat olfactory mucosa. However, this method did not allow accessing the data during the first 5 min of contact between the odorant and the mucosa; thus limiting the olfactory biological significance. Using a direct-injection mass spectrometry technique with a proton transfer reaction instrument (PTR-MS), we have been able, for the first time, to investigate the first moments of the enzymatic process of the metabolic capacity of ex vivo rat olfactory mucosa in real time. Using ethyl acetate as a model volatile odorous substrate, we demonstrated here for the first time that this odorant could be metabolised by an ex vivo olfactory mucosa within seconds, producing ethanol as metabolite.
Asunto(s)
Espectrometría de Masas/métodos , Monitoreo Fisiológico/métodos , Odorantes/análisis , Mucosa Olfatoria/metabolismo , Compuestos Orgánicos Volátiles/análisis , Acetatos/análisis , Acetatos/metabolismo , Animales , Masculino , Espectrometría de Masas/instrumentación , Técnicas de Cultivo de Órganos , Protones , Ratas Wistar , Compuestos Orgánicos Volátiles/metabolismoRESUMEN
Within the peripheral olfactory process, odorant metabolizing enzymes are involved in the active biotransformation of odorants, thus influencing the intensity and quality of the signal, but little evidence exists in humans. Here, we characterized the fast nasal metabolism of the food aroma pentane-2,3-dione in vivo and identified two resulting metabolites in the nasal-exhaled air, supporting the metabolizing role of the dicarbonyl/l-xylulose reductase. We showed in vitro, using the recombinant enzyme, that pentane-2,3-dione metabolism was inhibited by a second odorant (e.g., butanoic acid) according to an odorant-odorant competitive metabolic mechanism. Hypothesizing that such mechanism exists in vivo, pentane-2,3-dione, presented with a competitive odorant, both at subthreshold concentrations, was actually significantly perceived, suggesting an increase in its nasal availability. Our results, suggesting that odorant metabolizing enzymes can balance the relative detection of odorants in a mixture, in turn influencing the intensity of the signal, should be considered to better manage flavor perception in food.
Asunto(s)
Odorantes , Receptores Odorantes , Humanos , Pentanos , Receptores Odorantes/metabolismo , OlfatoRESUMEN
Addition of oenological tannins claims to have a positive impact on wine stability, protection from oxidation and likely sensory persistence. However, their role on red wine aroma during oxidation is controversial. The present study aims at investigating the effect of addition of oenological tannins on wine flavour (mainly aroma) before and after air exposure. Temporal Dominance of Sensations, a dynamic sensory evaluation, was coupled with a dynamic chemical measurement (nosespace analysis) using a Proton-Transfer-Reaction Mass-Spectrometer connected to the nasal cavity of 17 assessors. Results showed that the oxidation of a non-oaked Pinot Noir red wine decreases the fruity aroma dominance and increases the maderised and prune one. A contextual decrease of the fruity ethyl decanoate and increase of oxidative Strecker aldehydes are observed. Ellagitannins but not proanthocyanidins preserved perception of fruitiness and prevented increase of maderised notes. Moreover, ellagitannins increase the aroma persistence mainly in the non-oxidized wine.
Asunto(s)
Odorantes , Vino , Odorantes/análisis , Percepción , Taninos , Gusto , Vino/análisisRESUMEN
The present study aims at developing an analytical methodology which allows correlating sensory poles of chocolate to their chemical characteristics and, eventually, to those of the cocoa beans used for its preparation. Trained panelists investigated several samples of chocolate, and they divided them into four sensorial poles (characterized by 36 different descriptors) attributable to chocolate flavor. The same samples were analyzed by six different techniques: Proton Transfer Reaction-Time of Flight-Mass Spectrometry (PTR-ToF-MS), Solid Phase Micro Extraction-Gas Chromatography-Mass Spectroscopy (SPME-GC-MS), High-Performance Liquid Chromatography (HPLC) (for the quantification of eight organic acids), Ultra High Performance Liquid Chromatography coupled to triple-quadrupole Mass Spectrometry (UHPLC-QqQ-MS) for polyphenol quantification, 3D front face fluorescence Spectroscopy and Near Infrared Spectroscopy (NIRS). A multi-block classification approach (Sequential and Orthogonalized-Partial Least Squares - SO-PLS) has been used, in order to exploit the chemical information to predict the sensorial poles of samples. Among thirty-one test samples, only two were misclassified.
Asunto(s)
Cacao/química , Chocolate/análisis , Chocolate/clasificación , Análisis de los Alimentos/métodos , Cromatografía Líquida de Alta Presión , Análisis de los Alimentos/estadística & datos numéricos , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Análisis de los Mínimos Cuadrados , Espectrometría de Masas/métodos , Polifenoles/análisis , Microextracción en Fase Sólida , Espectrometría de Fluorescencia , Espectroscopía Infrarroja Corta , GustoRESUMEN
The feasibility of solid-state magic angle spinning (MAS) (31)P nuclear magnetic resonance (NMR) spectroscopy and (23)Na NMR spectroscopy to investigate both phosphates and Na(+) ions distribution in semi-hard cheeses in a non-destructive way was studied. Two semi-hard cheeses of known composition were made with two different salt contents. (31)P Single-pulse excitation and cross-polarization MAS experiments allowed, for the first time, the identification and quantification of soluble and insoluble phosphates in the cheeses. The presence of a relatively 'mobile' fraction of colloidal phosphates was evidenced. The detection by (23)Na single-quantum NMR experiments of all the sodium ions in the cheeses was validated. The presence of a fraction of 'bound' sodium ions was evidenced by (23)Na double-quantum filtered NMR experiments. We demonstrated that NMR is a suitable tool to investigate both phosphates and Na(+) ions distributions in cheeses. The impact of the sodium content on the various phosphorus forms distribution was discussed and results demonstrated that NMR would be an important tool for the cheese industry for the processes controls.
Asunto(s)
Queso , Fosfatos/química , Fósforo/química , Teoría Cuántica , Sodio/química , Intercambio Iónico , Iones , Espectroscopía de Resonancia MagnéticaRESUMEN
(23)Na NMR spectroscopy has been used to study the effects of Na(+) ion concentrations on the structure of 1% (w/w) iota-carrageenan systems, a natural gelling polysaccharide used as a thickener in the food industry. Rheological and (23)Na T(1) relaxation time measurements revealed that gel formation correlates with decreases in ion mobility over the range of 0-3% (w/w) sodium content. (23)Na single-quantum (SQ) and double-quantum-filtered (DQF) NMR experiments performed on these systems provided evidence for a 'bound' sodium ion fraction in a specifically ordered environment. These results have allowed us to propose a model for the carrageenan gelation mechanism in the presence of Na(+) ions.
Asunto(s)
Carragenina/química , Teoría Cuántica , Cloruro de Sodio/química , Espectroscopía de Resonancia Magnética/métodos , Estructura Molecular , Reología , Isótopos de SodioRESUMEN
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
RESUMEN
Odorant-metabolizing enzymes are critically involved in the clearance of odorant molecules from the environment of the nasal neuro-olfactory tissue to maintain the sensitivity of olfactory detection. Odorant metabolism may also generate metabolites in situ, the characterization and function of which in olfaction remain largely unknown. Here, we engineered and validated an ex vivo method to measure odorant metabolism in real-time. Glassware containing an explant of rat olfactory mucosa was continuously flushed with an odorant flow and was coupled to a proton transfer reaction-mass spectrometer for volatile compound analysis. Focusing on carboxylic esters and diketone odorants, we recorded the metabolic uptake of odorants by the mucosa, concomitantly with the release of volatile odorant metabolites in the headspace. These results significantly change the picture of real-time in situ odorant metabolism and represent a new step forward in the investigation of the function of odorant metabolites in the peripheral olfactory process. Our method allows the systematic identification of odorant metabolites using a validated animal model and permits the screening of olfactory endogenously produced chemosensory molecules.
Asunto(s)
Metabolómica/métodos , Mucosa Nasal/química , Odorantes/análisis , Animales , Ésteres/química , Cromatografía de Gases y Espectrometría de Masas , Cetonas/química , Modelos Animales , Ratas , Ratas WistarRESUMEN
Direct-injection mass spectrometry (DIMS) techniques have evolved into powerful methods to analyse volatile organic compounds (VOCs) without the need of chromatographic separation. Combined to chemometrics, they have been used in many domains to solve sample categorization issues based on volatilome determination. In this paper, different DIMS methods that have largely outperformed conventional electronic noses (e-noses) in classification tasks are briefly reviewed, with an emphasis on food-related applications. A particular attention is paid to proton transfer reaction mass spectrometry (PTR-MS), and many results obtained using the powerful PTR-time of flight-MS (PTR-ToF-MS) instrument are reviewed. Data analysis and feature selection issues are also summarized and discussed. As a case study, a challenging problem of classification of dark chocolates that has been previously assessed by sensory evaluation in four distinct categories is presented. The VOC profiles of a set of 206 chocolate samples classified in the four sensory categories were analysed by PTR-ToF-MS. A supervised multivariate data analysis based on partial least squares regression-discriminant analysis allowed the construction of a classification model that showed excellent prediction capability: 97% of a test set of 62 samples were correctly predicted in the sensory categories. Tentative identification of ions aided characterisation of chocolate classes. Variable selection using dedicated methods pinpointed some volatile compounds important for the discrimination of the chocolates. Among them, the CovSel method was used for the first time on PTR-MS data resulting in a selection of 10 features that allowed a good prediction to be achieved. Finally, challenges and future needs in the field are discussed.
Asunto(s)
Chocolate/análisis , Compuestos Orgánicos Volátiles/análisis , Calidad de los Alimentos , Espectrometría de Masas/métodos , SensaciónRESUMEN
With proton transfer reaction-mass spectrometry standard operating conditions, analysis of alcoholic beverages is an analytical challenge. Ethanol reacts with the primary ion H3 O+ leading to its depletion and to formation of ethanol-related ions and clusters, resulting in unstable ionization and in significant fragmentation of analytes. Different methods were proposed but generally resulted in lowering the sensitivity and/or complicating the mass spectra. The aim of the present study was to propose a simple, sensitive, and reliable method with fragmentation as low as possible, linearity within a realistic range of volatile organic compounds concentrations, and applicability to in vivo dynamic aroma release (nosespace) studies of wines. For in vitro analyses, a reference flask containing a hydro-alcoholic solution (10% ethanol) was permanently connected to the PTR-MS inlet in order to establish ethanol chemical ionization conditions. A low electric field strength to number density ratio E/N (80 Td) was used in the drift-tube. A stable reagent ion distribution was obtained with the primary protonated ethanol ion C2 H5 OH2+ accounting for more than 80% of the ionized species. The ethanol dimer (C2 H5 OH)2 H+ accounted for only 10%. Fragmentation of some aroma molecules important for white wine flavor (various esters, linalool, cis-rose oxide, 2-methylpropan-1-ol, 3-methylbutan-1-ol, and 2-phenylethanol) was studied from same ethanol content solutions connected alternatively with the reference solution to the instrument inlet. Linear dynamic range and limit of detection (LOD) were determined for ethyl hexanoate. Fragmentation of the protonated analytes was limited to a few ions of low intensity, or to specific fragment ions with no further fragmentation. Association and/or ligand switching reactions from ethanol clusters were only significant for the primary alcohols. Interpretation of the mass spectra was straightforward with easy detection of diagnostic ions. These results made this ethanol ionization method suitable for direct headspace analyses of model wines and to their nosespace analyses.
Asunto(s)
Espectrometría de Masas/métodos , Odorantes/análisis , Vino/análisis , Etanol/química , Humanos , Límite de Detección , Protones , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/químicaRESUMEN
Interactions between a well-characterized protein, beta-lactoglobulin, and two flavor compounds, beta-ionone and gamma-decalactone, were studied by 2D NMR spectroscopy. NMR spectra were recorded in aqueous solution (pH 2.0, 12 mM NaCl, 10% D(2)O) under conditions such that beta-lactoglobulin is present in a monomeric state. TOCSY and NOESY spectra were recorded on the protein and the complexes between protein and ligands. The spectra of the NH-CH(alpha) region showed the cross-signals due to the coupling between N- and C-bonded protons in the polypeptide backbone. The observed chemical shift variations in the presence of ligands can be assigned to changes in the protein conformation. It appears that the side chains of several amino acids are affected by binding of gamma-decalactone point into the central cavity (Leu46, Ile56, Met107, and Gln120), whereas binding of beta-ionone affects amino acids located in a groove near the outer surface of the protein (Leu104, Tyr120, and Asp129), as illustrated by molecular visualization. This NMR study provides precise information of the location of binding and confirms the existence of two different binding sites for aroma compounds on beta-lactoglobulin, which was suggested in previous competition studies by fluorometry or affinity chromatography and by structural information obtained from infrared spectroscopy.
Asunto(s)
Lactoglobulinas/metabolismo , Lactonas/metabolismo , Espectroscopía de Resonancia Magnética , Norisoprenoides , Terpenos/metabolismo , Sitios de Unión , Lactoglobulinas/química , Lactonas/química , Modelos Moleculares , Estructura Molecular , Odorantes/análisis , Conformación Proteica , Terpenos/químicaRESUMEN
This study aims to follow the kinetics of aroma compound release during model cheese consumption in order to clarify the relationships between flavor release and some oral parameters. Eight subjects participated in the study. Breathing, salivation, chewing, and swallowing were monitored during the eating process. Temporal nosespace analyses were performed using on-line atmospheric pressure ionization-mass spectrometry (API-MS) and off-line solid-phase Micro extraction-gas chromatography-mass spectrometry (SPME-GC-MS). Flavor release profiles were obtained only for ethyl hexanoate, heptan-2-one, and heptan-2-ol. Among them, only the concentrations of ethyl hexanoate and heptan-2-one could be determined by API-MS. Absence of competition between the aroma compounds was checked for both techniques. In-nose maximum concentration (C(max)) varied with aroma compounds. However, C(max) was reached at the same time (T(max)) for the three compounds. Interindividual differences were observed for most of the parameters studied and for all of the aroma compounds. They were related to the interindividual differences among the oral parameters. The aroma release parameters C(max) and AUC (area under the curve) could be related to respiratory and masticatory parameters. In most cases, the same relationships were observed whatever the nature of the aroma compound.
Asunto(s)
Queso/análisis , Fenómenos Fisiológicos del Sistema Digestivo , Ingestión de Alimentos , Odorantes/análisis , Deglución , Electromiografía , Humanos , Espectrometría de Masas , Masticación , Respiración , SalivaciónRESUMEN
This study deals with the release kinetics of nonvolatile compounds (NVC) (leucine, phenylalanine, glutamic acid, citric acid, lactic acid, propanoic acid, sodium, potassium, calcium, magnesium, chloride, and phosphates) during the eating of a model cheese and the relationships to some oral (salivary and masticatory) parameters. The aroma release has previously been characterized in similar conditions [Pionnier, E.; Chabanet, C.; Mioche, L.; Le Quéré, J.-L.; Salles, C. J. Agric. Food Chem. 2004, 52, xxx-xxx (1)]. Saliva samples were collected from the tongues of eight assessors at different times during and after the chewing sequence. Atmospheric pressure ionization-mass spectrometry and/or high-performance liquid chromatography analyses have been performed on these samples in order to quantify the 12 NVC released in saliva. The maximum concentration (C(max)) in saliva varied significantly according to the compound. However, there was no significant effect of the compound on the time to reach maximum concentration (T(max)). Interindividual differences were observed for most of the parameters and for all of the NVC studied. The parameters extracted from the release profiles of the NVC were closely correlated. High T(max) and AUC (area under the curve) values could be related to high chewing time and low saliva flow rates, low chewing rates, low masticatory performances, and low swallowing rates.
Asunto(s)
Queso/análisis , Fenómenos Fisiológicos del Sistema Digestivo , Ingestión de Alimentos , Saliva/química , Aminoácidos/análisis , Cloruros/análisis , Cromatografía Líquida de Alta Presión , Citratos/análisis , Electromiografía , Humanos , Ácido Láctico/análisis , Espectrometría de Masas , Masticación , Minerales/análisisRESUMEN
An atmospheric pressure chemical ionization ion-trap mass spectrometer was set up for the on-line analysis of aroma compounds. This instrument, which has been successfully employed for some years in several in vitro and in vivo flavour release studies, is described for the first time in detail. The ion source was fashioned from polyether ether ketone and operated at ambient pressure and temperature making use of a discharge corona pin facing coaxially the capillary ion entrance of the ion-trap mass spectrometer. Linear dynamic ranges (LDR), limits of detection (LOD) and other analytical characteristics have been re-evaluated. LDRs and LODs have been found fully compatible with the concentrations of aroma compounds commonly found in foods. Thus, detection limits have been found in the low ppt range for common flavouring aroma compounds (for example 5.3 ppt (0.82 ppbV) for ethyl hexanoate and 4.8 ppt (1.0 ppbV) for 2,5-dimethylpyrazine). This makes the instrument applicable for in vitro and in vivo aroma release investigations. The use of dynamic sensory techniques such as the temporal dominance of sensations (TDS) method conducted simultaneously with in vivo aroma release measurements allowed to get some new insights in the link between flavour release and flavour perception.
RESUMEN
For the on-line monitoring of flavour compound release, atmospheric pressure chemical ionization (APCI) and proton transfer reaction (PTR) combined to mass spectrometry (MS) are the most often used ionization technologies. APCI-MS was questioned for the quantification of volatiles in complex mixtures, but direct comparisons of APCI and PTR techniques applied on the same samples remain scarce. The aim of this work was to compare the potentialities of both techniques for the study of in vitro and in vivo flavour release. Aroma release from flavoured aqueous solutions (in vitro measurements in Teflon bags and glass vials) or flavoured candies (in vivo measurements on six panellists) was studied using APCI- and PTR-MS. Very similar results were obtained with both techniques. Their sensitivities, expressed as limit of detection of 2,5-dimethylpyrazine, were found equivalent at 12 ng/l air. Analyses of Teflon bag headspace revealed a poor repeatability and important ionization competitions with both APCI- and PTR-MS, particularly between an ester and a secondary alcohol. These phenomena were attributed to dependency on moisture content, gas/liquid volume ratio, proton affinities and product ion distribution, together with inherent drawbacks of Teflon bags (adsorption, condensation of water and polar molecules). Concerning the analyses of vial headspace and in vivo analyses, similar results were obtained with both techniques, revealing no competition phenomena. This study highlighted the equivalent performances of APCI-MS and PTR-MS for in vitro and in vivo flavour release investigations and provided useful data on the problematic use of sample bags for headspace analyses.
Asunto(s)
Dulces/análisis , Espectrometría de Masas/métodos , Odorantes/análisis , Compuestos Orgánicos Volátiles/análisis , Área Bajo la Curva , Femenino , Humanos , Cinética , Límite de Detección , MasculinoRESUMEN
In hard cooked cheeses, any interactions between the thermophilic starters as they grow during the cheese-making are critical, since they modify bacterial growth kinetics and acidification kinetics, so affecting the ripening process and the final characteristics of the cheese. Twenty-four experimental hard cooked cheeses were made under controlled conditions, the milk being inoculated with various combinations of thermophilic strains of Streptococcus thermophilus, Lactobacillus helveticus and Lactobacillus delbrueckii. Over the first day of manufacturing we recorded a wide range of different growth kinetics for each starter species used, and a wide range of pH kinetics, depending on the starter combination. Most of the bacterial variability could be statistically explained by the nature, quantity, and/or presence or absence of the different strains inoculated. Four main interactions between the three species were evidenced during cheese-making. There was antagonism between L. helveticus and L. delbrueckii. The lactobacilli had a positive effect on S. thermophilus, which was reciprocal for L. helveticus. L. helveticus had a negative effect on S. thermophilus cultivability. And the combination of S. thermophilus inoculated in large quantities and L. helveticus strain H2 had a negative effect on the growth of the L. delbrueckii strain D2. While the positive effect of L. delbrueckii on S. thermophilus probably corresponds to interactions in milk that have already been described and published, the other interactions were hitherto unknown. These interactions are of major importance for the growth kinetics of streptococci and thermophilic lactobacilli during cheese-making.