Relationships between the inhibitory efficacy and physicochemical properties of six organic acids and monolaurin against Bacillus weihenstephanensis KBAB4 growth in liquid medium.

Organic acids are widely used in foodstuffs to inhibit pathogen and spoiler growth. In this study, six organic acids (acetic, lactic, propionic, phenyllactic, caprylic, and lauric acid) and monolaurin were selected based on their physicochemical properties: their molecular structure (carbon chain length), their lipophilicity (logP), and their ability to dissociate in a liquid environment (pKa). The relation between these physicochemical properties and the inhibitory efficacy against B. weihenstephanensis KBAB4 growth was evaluated. After assessing the active form of these compounds against the strain (undissociated, dissociated or both forms), their MIC values were estimated in nutrient broth at pH 6.0 and 5.5 using two models (Lambert & Pearson, 2000; Luong, 1985). The use of two models highlighted the mode of action of an antibacterial compound in its environment, thanks to the additional estimation of the curve shape α or the Non-Inhibitory Concentration (NIC). The undissociated form of the tested acids is responsible for growth inhibition, except for lauric acid and monolaurin. Moreover, long-carbon chain acids have lower estimated MICs, compared to short-chain acids. Thus, the inhibitory efficacy of organic acids is strongly related to their carbon chain length and lipophilicity. Lipophilicity is the main mechanism of action of a membrane-active compound, it can be favored by long chain structure or high pKa in an acid environment like food.

Impact of carbon dioxide on the radial growth of fungi isolated from dairy environment.

In dairy industry, filamentous fungi are used as adjunct cultures in fermented products for their technological properties but they could also be responsible for food spoilage and mycotoxin production. The consumer demands about free-preservative products has increased in recent years and lead to develop alternative methods for food preservation. Modified Atmosphere Packaging (MAP) can inhibit fungal growth and therefore increase the food product shelf-life. This study aimed to evaluate radial growth as a function of CO2 and more particularly carbonic acid for fourteen adjuncts and/or fungal spoiler isolated from dairy products or dairy environment by using predictive mycology tools. The impact of the different chemical species linked to CO2 (notably carbonic acid) were study because it was reported previously that undissociated carbonic acid impacted bacterial growth and bicarbonates ions were involved in modifications of physiological process of fungal cells. A significant diversity in the responses of selected strains was observed. Mucor circinelloides had the fastest growth rates (µ > 11 mm. day-1) while Bisifusarium domesticum, Cladosporium herbarum and Penicillium bialowiezense had the slowest growth rates (µ < 1 mm. day-1). Independently of the medium pH, the majority of strains were sensitive to total carbonic acid. In this case, it was not possible to conclude if CO2 active form was gaseous or aqueous so modeling were performed as a function of CO2 percentage. Only Geotrichum candidum and M. circinelloides strains were sensitive to undissociated carbonic acid. Among the fourteen strains, P. bialowiezense was the less sensitive strain to CO2, no growth was observed at 50% of CO2 only for this strain. M. lanceolatus was the less sensitive strain to CO2, the CO250 which reduce the growth rates by 50% was estimated at 138% of CO2. Low CO2 percentage improved the growth of Penicillium expansum, Penicillium roqueforti and Paecilomyces niveus. Mathematical models (without and with optimum) were suggested to describe the impact of CO2 percentage or undissociated carbonic acid concentration on fungal growth rate.

Impact of sodium chloride and carbon dioxide on conidial germination and radial growth of Penicillium camemberti.

Penicillium camemberti is a domesticated species adapted to the dairy environment, which is used as adjunct cultures to ripen soft cheeses. A recent population genomics analysis on P. camemberti revealed that P. camemberti is a clonal lineage with two varieties almost identical genetically but with contrasting phenotypes in terms of growth, color, mycotoxin production and inhibition of contaminants. P. camemberti variety camemberti is found on Camembert and Brie cheeses, and P. camemberti variety caseifulvum is mainly found on other cheeses like Saint-Marcellin and Rigotte de Condrieu. This study aimed to evaluate the impact of water activity (aw) reduced by sodium chloride (NaCl) and the increase of carbon dioxide (CO2) partial pressure, on conidial germination and growth of two varieties of P. camemberti: var. Camemberti and var. Caseifulvum. Mathematical models were used to describe the responses of P. camemberti strains to both abiotic factors. The results showed that these genetically distant strains had similar responses to increase in NaCl and CO2 partial pressure. The estimated cardinal values were very close between the strains although all estimated cardinal values were significantly different (Likelihood ratio tests, pvalue = 0.05%). These results suggest that intraspecific variability could be more exacerbated during fungal growth compared with conidial germination, especially in terms of macroscopic morphology. Indeed, var. Caseifulvum seemed to be more sensitive to an increase of CO2 partial pressure, as shown by the fungal morphology, with the occurrence of irregular outgrowths, while the morphology of var. Camemberti remains circular. These data could make it possible to improve the control of fungal development as a function of salt and carbon dioxide partial pressure. These abiotic factors could serve as technological barriers to prevent spoilage and increase the shelf life of cheeses. The present data will allow more precise predictions of fungal proliferation as a function of salt and carbon dioxide partial pressure, which are significant technological hurdles in cheese production.

Synergistic interaction between pH and NaCl in the limits of germination and outgrowth of Clostridium sporogenes and Group I Clostridium botulinum vegetative cells and spores after heat treatment.

Group I Clostridium botulinum and Clostridium sporogenes are physiologically and genetically closely related. Both are widely distributed in the environment and can cause foodborne botulism. In this work, a physiological study was conducted with 37 isolates from spoiled canned food and five referenced strains of C. sporogenes (three isolates) and Group I C. botulinum (two isolates). Growth limits of vegetative cells were established as a function of pH and NaCl concentration in PYG modified medium (PYGm) at 30 °C for 48 days. The heat resistance of the spores was studied for 2 min and 10 min at 102 °C and 110 °C. This physiological study (pH, NaCl growth limits and heat resistance) allowed the selection of 14 isolates of C. sporogenes (twelve isolates) and Group I C. botulinum (two isolates) representative of the diversity found. This panel of 14 selected isolates (11 isolated from spoiled canned food and three reference strains), were whole genome sequenced, but no association of physiological and genetic characteristics could be detected. Finally, we studied the ability of spores to germinate and grow from 5 isolates (four C. sporogenes and one Group I C. botulinum), under stress conditions generated by pH and NaCl following a low intensity heat treatment. The accumulation of these 3 stresses creates synergies that will strongly reduce the probability of spore growth in pH and salt conditions where they usually proliferate. The effect is progressive as the conditions become drastic: the number of decimal reduction observed increases translating a probability of growth which decreases. This study provides a better understanding of the behaviour of C. sporogenes and Group I C. botulinum isolates and shows how the combination of pH, NaCl and heat treatment can help prevent or minimise foodborne botulism outbreaks.

The synergic interaction between environmental factors (pH and NaCl) and the physiological state (vegetative cells and spores) provides new possibilities for optimizing processes to manage risk of C. sporogenes spoilage.

Clostridium sporogenes has been widely used as a surrogate for proteolytic C. botulinum for validating thermal processes in low-acid cans. To limit the intensity of heat treatments, industrials must use other ways of control as an association of acidic and saline environment after a low heat treatment. The probability of growth of pH (7-4.4), sodium chloride concentration (0-11%) and heat treatment (80°C-10 min; 100°C-1.5 min and 5.2 min) were studied on C. sporogenes PA 3679 spores and vegetative cells. Vegetative cells or heat-treated spores were inoculated in PYGm broth at 30 °C for 48 days in anaerobic conditions. Vegetative cells growth (pH 4.6-pH 4.5; 7%-8% NaCl) range is larger than the spore one (pH 5.2-pH 5.0; 6%-7% NaCl). Spores germination and outgrowth rage is decreased if the spores are heat-treated at 100 °C for 1.5 min (pH 5.5-5.3; 4%-5% NaCl) and 5.2 min (pH 5.7-5.3; 4%-5% NaCl). The C. sporogenes PA 3679 spores germination and outgrowth is impacted by their physiological state. The synergic interaction between environmental factors (pH and NaCl) and the physiological state (vegetative cells and spores) opening new possibilities for optimizing food formulation processes to manage the risks of C. sporogenes spoilage.

Differentiation of Vegetative Cells into Spores: a Kinetic Model Applied to Bacillus subtilis.

Spore-forming bacteria are natural contaminants of food raw materials, and sporulation can occur in many environments from farm to fork. In order to characterize and to predict spore formation over time, we developed a model that describes both the kinetics of growth and the differentiation of vegetative cells into spores. The model is based on a classical growth model and enables description of the kinetics of sporulation with the addition of three parameters specific to sporulation. Two parameters are related to the probability of each vegetative cell to commit to sporulation and to form a spore, and the last one is related to the time needed to form a spore once the cell is committed to sporulation. The goodness of fit of this growth-sporulation model was assessed using growth-sporulation kinetics at various temperatures in laboratory medium or in whey for Bacillus subtilis, Bacillus cereus, and Bacillus licheniformis The model accurately describes the kinetics in these different conditions, with a mean error lower than 0.78 log10 CFU/ml for the growth and 1.08 log10 CFU/ml for the sporulation. The biological meaning of the parameters was validated with a derivative strain of Bacillus subtilis 168 which produces green fluorescent protein at the initiation of sporulation. This model provides physiological information on the spore formation and on the temporal abilities of vegetative cells to differentiate into spores and reveals the heterogeneity of spore formation during and after growth.IMPORTANCE The growth-sporulation model describes the progressive transition from vegetative cells to spores with sporulation parameters describing the sporulation potential of each vegetative cell. Consequently, the model constitutes an interesting tool to assess the sporulation potential of a bacterial population over time with accurate parameters such as the time needed to obtain one resistant spore and the probability of sporulation. Further, this model can be used to assess these data under various environmental conditions in order to better identify the conditions favorable for sporulation regarding the time to obtain the first spore and/or the concentrations of spores which could be reached during a food process.

Walking dead: Permeabilization of heat-treated Geobacillus stearothermophilus ATCC 12980 spores under growth-preventing conditions.

Although heat treatment is probably the oldest and the most common method used to inactivate spores in food processes, the specific mechanism of heat killing of spores is still not fully understood. The purpose of this study is to investigate the evolution of the permeabilization and the viability of heat-treated spores during storage under growth-preventing conditions. Geobacillus stearothermophilus spores were heat-treated under various conditions of temperature and pH, and then stored under conditions of temperature and pH that prevent growth. Spore survival was evaluated by count plating immediately after heat treatment, and then during storage over a period of months. Flow cytometry analyses were performed to investigate the Syto 9 permeability of heat-treated spores. Sub-lethally heat-treated spores of G. stearothermophilus were physically committed to permeabilization after heat treatment. However, prolonged heat treatment may abolish the spore permeabilization and block heat-treated spores in the refractive state. However, viability loss and permeabilization during heat treatment seem to be two different mechanisms that occur independently, and the loss of permeabilization properties takes place at a much slower rate than spore killing. Under growth-preventing conditions, viable heat-treated spores presumably lose their viability due to the permeabilization phenomena, which makes them more susceptible to the action of adverse conditions precluding growth.

Modelling the effect of water activity reduction by sodium chloride or glycerol on conidial germination and radial growth of filamentous fungi encountered in dairy foods.

Water activity (aw) is one of the most influential abiotic factors affecting fungal development in foods. The effects of aw reduction on conidial germination and radial growth are generally studied by supplementing culture medium with the non-ionic solute glycerol despite food aw can also depend on the concentration of ionic solutes such as sodium chloride (NaCl). The present study aimed at modelling and comparing the effects of aw, either modified using NaCl or glycerol, on radial growth and/or conidial germination parameters for five fungal species occurring in the dairy environment. The estimated cardinal values were then used for growth prediction and compared to growth kinetics observed on commercial fresh cheese. Overall, as compared to glycerol, NaCl significantly increased the fungistatic effect resulting from aw reduction by extending latency and/or reducing radial growth rates of Paecilomyces niveus, Penicillium brevicompactum, Penicillium expansum and Penicillium roqueforti but not of Mucor lanceolatus. Besides, NaCl significantly reduced aw range for conidial germination and delayed median germination time of P. expansum but not of P. roqueforti. Despite these observations, cardinal aw values obtained on glycerol-medium yielded similar predictions of radial growth and germination time in commercial fresh cheese as those obtained with NaCl. Thus, it indicates that, for the studied species and aw range used for model validation, the use of NaCl instead of glycerol as a aw depressor had only limited impact for fungal behavior prediction.

Effect of temperature, pH, and water activity on Mucor spp. growth on synthetic medium, cheese analog and cheese.

The Mucor genus includes a large number of ubiquitous fungal species. In the dairy environment, some of them play a technological role providing typical organoleptic qualities to some cheeses while others can cause spoilage. In this study, we compared the effect of relevant abiotic factors for cheese production on the growth of six strains representative of dairy technological and contaminant species as well as of a non cheese related strain (plant endophyte). Growth kinetics were determined for each strain in function of temperature, water activity and pH on synthetic Potato Dextrose Agar (PDA), and secondary models were fitted to calculate the corresponding specific cardinal values. Using these values and growth kinetics acquired at 15 °C on cheese agar medium (CA) along with three different cheese types, optimal growth rates (µopt) were estimated and consequently used to establish a predictive model. Contrarily to contaminant strains, technological strains showed higher µopt on cheese matrices than on PDA. Interestingly, lag times of the endophyte strain were strongly extended on cheese related matrices. This study offers a relevant predictive model of growth that may be used for better cheese production control but also raises the question of adaptation of some Mucor strains to the cheese.

Effect of pH on Thermoanaerobacterium thermosaccharolyticum DSM 571 growth, spore heat resistance and recovery.

Thermophilic spore-forming bacteria are potential contaminants in several industrial sectors involving high temperatures (40-65 °C) in the manufacturing process. Among those thermophilic spore-forming bacteria, Thermoanaerobacterium thermosaccharolyticum, called "the swelling canned food spoiler", has generated interest over the last decade in the food sector. The aim of this study was to investigate and to model pH effect on growth, heat resistance and recovery abilities after a heat-treatment of T. thermosaccharolyticum DSM 571. Growth and sporulation were conducted on reinforced clostridium media and liver broth respectively. The highest spore heat resistances and the greatest recovery ability after a heat-treatment were obtained at pH condition allowing maximal growth rate. Growth and sporulation boundaries were estimated, then models using growth limits as main parameters were extended to describe and quantify the effect of pH on recovery of injured spores after a heat-treatment. So, cardinal values were used as a single set of parameters to describe growth, sporulation and recovery abilities. Besides, this work suggests that T. thermosaccharolyticum preserve its ability for germination and outgrowth after a heat-treatment at a low pH where other high resistant spore-forming bacteria like Geobacillus stearothermophilus are unable to grow.

Die another day: Fate of heat-treated Geobacillus stearothermophilus ATCC 12980 spores during storage under growth-preventing conditions.

Geobacillus stearothermophilus spores are recognized as one of the most wet-heat resistant among aerobic spore-forming bacteria and are responsible for 35% of canned food spoilage after incubation at 55 °C. The purpose of this study was to investigate and model the fate of heat-treated survivor spores of G. stearothermophilus ATCC 12980 in growth-preventing environment. G. stearothermophilus spores were heat-treated at four different conditions to reach one or two decimal reductions. Heat-treated spores were stored in nutrient broth at different temperatures and pH under growth-preventing conditions. Spore survival during storage was evaluated by count plating over a period of months. Results reveal that G. stearothermophilus spores surviving heat treatment lose their viability during storage under growth-preventing conditions. Two different subpopulations were observed during non-thermal inactivation. They differed according to the level of their resistance to storage stress, and the proportion of each subpopulation can be modulated by heat treatment conditions. Finally, tolerance to storage stress under growth-preventing conditions increases at refrigerated temperature and neutral pH regardless of heat treatment conditions. Such results suggest that spore inactivation due to heat treatment could be completed by storage under growth-preventing conditions.

Extending the gamma concept to non-thermal inactivation: a dynamic model to predict the fate of Salmonella during the dried sausages process.

The process of dried fermented sausages is recognized to be favourable to the reduction of the Salmonella population. The objective of this study was to develop a model describing the evolution of Salmonella during the fabrication process of dried sausages and to optimize the food formulation to prevent pathogen presence at the end of the process. An experimental design was set to investigate the effects of the fermentation and drying process for several formulations, taking into account the type of starter culture, the sodium chloride concentration, the dextrose and lactose concentration on the Salmonella Typhimurium strain behaviour. A growth-inactivation model based on the gamma concept was then developed to quantify Salmonella behaviour in dynamic process conditions of temperature, pH, lactic acid and water activity. This behaviour was characterized by a first growth step, followed by an inactivation step. The Salmonella fate was well described by the model in terms of population size variation and transition from growth to inactivation. The Salmonella behaviour was influenced by the initial sugar concentration and the starter type but not by sodium chloride content. This model can be a valuable tool to design the food process and formulation to control Salmonella.

mRNA biomarkers selection based on Partial Least Square algorithm in order to further predict Bacillus weihenstephanensis acid resistance.

In order to integrate omics data to quantitative microbiological risk assessment in foods, gene expressions may serve as bacterial behaviour biomarkers. In this study an integrative approach encompassing predictive modelling and mRNAs quantifications, was followed to select molecular biomarkers to further predict the acid resistance of Bacillus weihenstephanensis. A multivariate analysis was performed to correlate the acid bacterial resistance and the gene expression of vegetative cells with or without exposure to stressing conditions. This mathematical method provides the advantage to take gene expressions and their interactions into account. The use of the Partial Least Squares algorithm allowed the selection of nine genes as acid resistance biomarkers among thirty targeted genes. According to their involvement in the general acid stress response of Bacillus, these genes were assigned to three different biological modules namely, metabolic rearrangements, general stress response and oxidative stress response. The oxidative stress response appeared as the major activated biological module in B. weihenstephanensis cells submitted to acid stress conditions. Furthermore, as a firstly described model, the developed concept showed promising results to further be used to predict bacterial resistance using gene expression. Thus, this study underlines the possibility to integrate the bacterial physiology state, using omics biomarkers, into bacterial behaviour modelling and provide mechanistic understanding in acid bacterial resistance mechanisms.

Modeling the behavior of Geobacillus stearothermophilus ATCC 12980 throughout its life cycle as vegetative cells or spores using growth boundaries.

Geobacillus stearothermophilus is recognized as one of the most prevalent micro-organism responsible for flat sour in the canned food industry. To control these highly resistant spore-forming bacteria, the heat treatment intensity could be associated with detrimental conditions for germination and outgrowth. The purpose of this work was to study successively the impact of temperature and pH on the growth rate of G. stearothermophilus ATCC 12980, its sporulation ability, its heat resistance in response to various sporulation conditions, and its recovery ability after a heat treatment. The phenotypic investigation was carried out at different temperatures and pHs on nutrient agar and the heat resistance was estimated at 115 °C. The greatest spore production and the highest heat resistances were obtained at conditions of temperature and pH allowing maximal growth rate. The current observations also revealed that growth, sporulation and recovery boundaries are close. Models using growth boundaries as main parameters were extended to describe and quantify the effect of temperature and pH throughout the life cycle of G. stearothermophilus as vegetative cells or as spore after a heat treatment and during recovery.

An integrative approach to identify Bacillus weihenstephanensis resistance biomarkers using gene expression quantification throughout acid inactivation.

The aim of this study was to define an integrative approach to identify resistance biomarkers using gene expression quantification and mathematical modelling. Mid-exponentially growing cells were transferred into acid conditions (BHI, pH 4.6) to obtain inactivation kinetics, performed in triplicate. The inactivation curve was fitted with a mixed Weibull model. This model allowed to differentiate two subpopulations with various acid resistances among the initial population. In parallel, differential gene expression was quantified by RT-qPCR. While narL was down-regulated throughout acid inactivation, sigB and katA were up-regulated. sigB expression up-regulation peak was correlated to the less resistant subpopulation when katA up-regulation, was correlated to the more resistant subpopulation. Moreover, differences in population structure were highlighted between each replicate. The higher proportion of the more resistant subpopulation was linked to a higher katA gene expression. These results suggest that sigB and katA might be used as different types of biomarkers, for instance to track moderate and high acid-resistance, respectively. The use of this approach combining RT-qPCR and predictive modelling to track cellular biomarker variations appears as an interesting tool to take into account physiological cell responses into mathematical modelling, allowing an accurate prediction of microbial behaviour.

Modeling heat resistance of Bacillus weihenstephanensis and Bacillus licheniformis spores as function of sporulation temperature and pH.

Although sporulation environmental factors are known to impact on Bacillus spore heat resistance, they are not integrated into predictive models used to calculate the efficiency of heating processes. This work reports the influence of temperature and pH encountered during sporulation on heat resistance of Bacillus weihenstephanensis KBAB4 and Bacillus licheniformis AD978 spores. A decrease in heat resistance (δ) was observed for spores produced either at low temperature, at high temperature or at acidic pH. Sporulation temperature and pH maximizing the spore heat resistance were identified. Heat sensitivity (z) was not modified whatever the sporulation environmental factors were. A resistance secondary model inspired by the Rosso model was proposed. Sporulation temperatures and pHs minimizing or maximizing the spore heat resistance (T(min(R)), T(opt(R)), T(max(R)), pH(min(R)) and pH(opt(R))) were estimated. The goodness of the model fit was assessed for both studied strains and literature data. The estimation of the sporulation temperature and pH maximizing the spore heat resistance is of great interest to produce spores assessing the spore inactivation in the heating processes applied by the food industry.

Sporulation boundaries and spore formation kinetics of Bacillus spp. as a function of temperature, pH and a(w).

Sporulation niches in the food chain are considered as a source of hazard and are not clearly identified. Determining the sporulation environmental boundaries could contribute to identify potential sporulation niches. Spore formation was determined in a Sporulation Mineral Buffer. The effect of incubation temperature, pH and water activity on time to one spore per mL, maximum sporulation rate and final spore concentration was investigated for a Bacillus weihenstephanensis and a Bacillus licheniformis strain. Sporulation boundaries of B. weihenstephanensis and of B. licheniformis were similar to, or included within, the range of temperatures, pH and water activities supporting growth. For instance, sporulation boundaries of B. weihenstephanensis were evaluated at 5°C, 35°C, pH 5.2 and a(w) 0.960 while growth boundaries were observed at 5°C, 37°C, pH 4.9 and a(w) 0.950. Optimum spore formation was determined at 30°C pH 7.2 for B. weihenstephanensis and at 45°C pH 7.2 for B. licheniformis. Lower temperatures and pH delayed the sporulation process. For instance, the time to one spore per mL was tenfold longer when sporulation occurred at 10°C and 20°C, for each strain respectively, than at optimum sporulation temperature. The relative effect of temperature and pH on sporulation rates and on growth rates is similar. This work suggests that the influence of environmental factors on the quantitative changes in sporulation boundaries and rates was similar to their influence on changes in growth rate.

Effect of abiotic factors and culture media on the growth of cheese-associated Nectriaceae species.

Nectriaceae species have been described in various natural environments or as plant or human pathogens. Within this family, the Bisifusarium domesticum species is of particular interest for food mycologists as it is used for technological functions in various cheese productions. Moreover, it has only been isolated from the cheese environment so far and, until recently, was the only Nectriaceae species described in this food product. Recently, four novel cheese-associated Nectriaceae species have been described, including two associated to the Bisifusarium genus and two to a new genus, Longinectria gen. nov.. These observations raise questions concerning the potential adaptation of these species to the cheese environment. In this context, this study first focused on determining the impact of abiotic factors on the growth of isolates belonging to the five cheese-associated species (i.e. B. allantoides sp. nov., B. domesticum, B. penicilloides sp. nov., L. lagenoides gen. nov. sp. nov. and L. verticilliforme gen. nov. sp. nov.) but also included phylogenetically close species. To do so, fungal growth kinetics in liquid medium (Potato Dextrose Broth) were determined by laser nephelometry at different temperatures, pH and water activities using NaCl as a depressor. Growth modeling was then performed to estimate cardinal values for each abiotic factor. Secondly, fungal growth was also evaluated on Potato Dextrose Agar (synthetic medium), cheese agar (cheese-mimicking medium) and Raclette de Savoie cheese (actual cheese). Our results clearly highlighted physiological differences in growth characteristics between the studied cheese-associated Nectriaceae spp. and the "non-cheese" species which could suggest, for the former, an adaptation to this food matrix. Indeed, regarding the impact of the tested abiotic factors, statistical analyses confirmed this dichotomy, with for example the lowest optimal temperatures estimated for the cheese-associated species (Topt 19.1-23.1 °C) while the other Bisifusarium species exhibited the highest optimal temperatures (Topt 26.1-36.2 °C). As for the impact of growth media, radial growth measurements highlighted that B. domesticum was the least affected species for growth on Raclette de Savoie and even grew faster on cheese agar than on synthetic medium confirming its strong adaptation to the cheese environment.

Impact of water activity on the radial growth of fungi in a dairy environment.

Filamentous fungi are used in the dairy industry as adjunct cultures in fermented products, but can also lead to food spoilage, waste and economic losses. The control of filamentous fungi with abiotic factors contributes to longer food shelf life and prevention of fungal spoilage. One of the main abiotic factors for controlling fungal growth in foods is water activity (aw). This study aimed to evaluate radial growth as a function of aw for sixteen fungal adjuncts and/or spoilers isolated from dairy products or a dairy environment. Glycerol (a non-ionic compound) and sodium chloride (NaCl, an ionic compound) were used to adjust the aw of culture media. This study showed significant diversity in the responses of the tested fungal strains as a function of medium aw. The growth response of Penicillium bialowiezense and Sporendonema casei was binary, with no clear decrease of growth rate until the growth limit, when the aw was reduced. For the strains of Bisifusarium domesticum, Mucor circinelloides and Penicillium camemberti, a decrease of aw had the same impact on radial growth rate regardless of the aw belonging to their growth range. For the strains of Aspergillus flavus, Cladosporium herbarum, Geotrichum candidum, Mucor lanceolatus, Penicillium expansum, Penicillium fuscoglaucum, Penicillium nalgiovense, Paecilomyces niveus, Penicillium roqueforti, Penicillium solitum and Scopulariopsis asperula, the impact of a decrease in aw was more pronounced at high aw than at low aw. A mathematical model was suggested to describe this impact on the radial growth rate. For all tested species, radial growth was more sensitive to NaCl than glycerol. The ionic strength of NaCl mainly explains the difference in the effects of the two solutes.

A Bayesian Approach to Describe and Simulate the pH Evolution of Fresh Meat Products Depending on the Preservation Conditions.

Measuring the pH of meat products during storage represents an efficient way to monitor microbial spoilage, since pH is often linked to the growth of several spoilage-associated microorganisms under different conditions. The present work aimed to develop a modelling approach to describe and simulate the pH evolution of fresh meat products, depending on the preservation conditions. The measurement of pH on fresh poultry sausages, made with several lactate formulations and packed under three modified atmospheres (MAP), from several industrial production batches, was used as case-study. A hierarchical Bayesian approach was developed to better adjust kinetic models while handling a low number of measurement points. The pH changes were described as a two-phase evolution, with a first decreasing phase followed by a stabilisation phase. This stabilisation likely took place around the 13th day of storage, under all the considered lactate and MAP conditions. The effects of lactate and MAP on pH previously observed were confirmed herein: (i) lactate addition notably slowed down acidification, regardless of the packaging, whereas (ii) the 50%CO2-50%N2 MAP accelerated the acidification phase. The Bayesian modelling workflow-and the script-could be used for further model adaptation for the pH of other food products and/or other preservation strategies.