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Predicting the microbial safety of food products stored in modified atmosphere packaging implies taking into account the effect of oxygen reduction on microbial growth. According to their respiratory-type, the micro-organisms are not impacted similarly by the oxygen concentration. The aim of this article was to quantify and model the oxygen effect on the growth rates of 5 bacterial species: Listeria monocytogenes and Bacillus weihenstephanensis (facultative anaerobic), Pseudomonas fluorescens (strict aerobic), Clostridium perfringens and Clostridium sporogenes (strict anaerobic). The results showed the oxygen concentration doesn't modify the behavior of both facultative anaerobic strains. The growth rate of P. fluorescens decreased with the oxygen concentration, but the effect is only noticeable when the oxygen concentration fell below 3% in the gaseous phase. Conversely, the oxygen acted as a growth inhibitor for both Clostridium species. But total inhibition is reached only for 3.26% and 6.61% respectively for C. sporogenes and C. perfringens. Two models have been fitted for both respiratory-types, the first is the Monod model considering oxygen as a substrate for growth, and the second is the classic inhibitory model based on minimal inhibitory concentration.

The genetic diversity of two major yeast species found in cheese, Debaryomyces hansenii and Kluyveromyces marxianus, was analyzed within the yeast flora in French traditional cheesemaking. Based on the amplification of sequences separating long terminal repeat (LTR) retrotransposon sequences, a molecular typing method was developed for D. hansenii and K. marxianus. This method was applied to a total of 56 D. hansenii strains and 61 K. marxianus strains, mostly isolated during fermentation and ripening of traditional cheese from Normandy and Haute-Savoie (French Alps) regions. A total of 32 and 43 robust profiles were obtained for D. hansenii and K. marxianus, respectively. Cluster analysis confirmed the large genetic diversity already shown for D. hansenii and revealed an even larger diversity for K. marxianus. After its use with Saccharomyces cerevisiae, the inter-LTR PCR proved to be efficient to discriminate between strains of the two species, D. hansenii and K. marxianus, isolated from the same ecological niches, confirming the high intra-specific variability of species found in cheese. This strain typing could not correlate the analyzed strains with their origin, would it be the cheese type, the cheese-making facility or the cheese batch, showing a high discrimination power. The method described here will provide a fast and reliable tool for the biodiversity study of these two major cheese yeasts.

Predicting microbial safety of fresh products in modified atmosphere packaging implies to take into account the dynamic of O2, CO2 and N2 exchanges in the system and its effect on microbial growth. In this paper a mechanistic model coupling gas transfer and predictive microbiology was validated using dedicated challenge-tests performed on poultry meat, fresh salmon and processed cheese, inoculated with either Listeria monocytogenes or Pseudomonas fluorescens and packed in commercially used packaging materials (tray + lid films). The model succeeded in predicting the relative variation of O2, CO2 and N2 partial pressure in headspace and the growth of the studied microorganisms without any parameter identification. This work highlighted that the respiration of the targeted microorganism itself and/or that of the naturally present microflora could not be neglected in most of the cases, and could, in the particular case of aerobic microbes contribute to limit the growth by removing all residual O2 in the package. This work also confirmed the low sensitivity of L. monocytogenes toward CO2 while that of P. fluorescens permitted to efficiently prevent its growth by choosing the right combination of packaging gas permeability value and initial % of CO2 initially flushed in the pack.

Depending on environmental factors, the prediction of bacterial growth is made difficult by the complexity of foodstuff. Although the influence of temperature, pH, and water activity are usually taken into account, models have to be completed with the influence of acid mixture. Nine strains of Listeria spp., four Salmonella spp., one Staphylococcus aureus, one Escherichia coli, and Listeria innocua ATCC 33090 were used for this study to extend model proposed by [Le Marc, Y., Huchet, V., Bourgeois, C., Guyonnet, J., Mafart, P., Thuault, D., 2002. Modelling the growth kinetics of Listeria as a function of temperature, pH and organic acid concentration. International Journal of Food Microbiology 73, 219-237]. Derived from data of [Houtsma, P.C., Kusters, B.J., De Wit, J.C., Rombouts, F.M., Zwietering, M.H., 1994. Modelling growth rates of Listeria innocua as a function of lactate concentration. International Journal of Food Microbiology 24, 113-123] and our own data, the extended model described accurately different effects of addition of acid salts in the medium (decrease of water activity and pH, variation of undissociated weak acid form, and variation of synergetic effect between environmental factors). This previous model was implemented to describe the observed variability of behaviour of the different studied strains. alpha reflected the general behaviour of species (sensitiveness to low or high undissociated acid concentration), and MIC(U) reflected the various resistances of strains. From this simple model, a new model was built for describing the effects of concentrations of several mixed acids on bacterial growth rates. Simulations of growth were carried out from three acids mixtures by inputting parameter estimates previously obtained. Despite a very variable effect of investigated acids on growth, the new model yielded fair predictions.

The combined effects of temperature, pH and organic acids (lactic, acetic and propionic) on the growth kinetics of Listeria innocua ATCC 33090 were studied. First, a multiplicative model was built assuming independent effects of all environmental factors. Thus, the model was expanded by the inclusion of a novel term describing the effects of interactions on the growth/no growth limits. The proposed approach allows an accurate description of the boundary between growth and no growth of Listeria.

The aim of this study was to model the growth, in a model system, of toxigenic strains of Bacillus cereus as a function of temperature, pH and water activity. Optical density (OD) values were transformed into numbers of colony-forming units (CFU) by the use of a `calibrating' relation. The growth curves were then fitted to the Gompertz function which allowed the estimation of the lag-time ( λ ) and the growth rate ( μ ). These two parameters were then modelled according to the controlling factors which were pH and water activity at 20 and 30°C.

A study of the factors influencingClostridium tyrobutyricumgrowth is necessary to improve the control of butyric fermentation and avoid late blowing during cheese making. Lactic acid, one of the substrates of this fermentation is present in two optical forms in cheeses and their relative amounts may be a controlling factor. In order to determine whether these proportions could influenceC. tyrobutyricumgrowth, the micro-organism was inoculated in culture media containing different proportions of these isomers. In the racemic mix, the results show the consumption of 90% of D (−) and 25% of L (+) lactic acid during the exponential growth phase. When both isomers are present,C. tyrobutyricumpreferentially uses the D (−) lactic acid, and if only one of the two isomers is present its growth rate is three times higher with the D (−) than with the L (+) isomer.

Lactococcin, a bacteriocin produced by Lactococcus lactis subsp. lactis ADRIA 85LO30, was purified as a 2.3-2.4 kDa peptide. Six non-bacteriocin-producing (Bac-) and non-immune (Imm-) strains were isolated after curing experiments. These strains had in common the loss or modification of two plasmids: pOS4 (32 kb) and pOS5 (70 kb). By comparing pOS5 and several modified plasmids, a DNA region from pOS5 of about 10 kb, which was necessary for wild-type bacteriocin production and immunity, was identified.

An experimental protocol to validate secondary-model application to foods was suggested. Escherichia coli , Listeria monocytogenes , Bacillus cereus , Clostridium perfringens , and Salmonella were observed in various food categories, such as meat, dairy, egg, or seafood products. The secondary model validated in this study was based on the gamma concept, in which the environmental factors temperature, pH, and water activity (aw) were introduced as individual terms with microbe-dependent parameters, and the effect of foodstuffs on the growth rates of these species was described with a food- and microbe-dependent parameter. This food-oriented approach was carried out by challenge testing, generally at 15 and 10°C for L. monocytogenes , E. coli , B. cereus , and Salmonella and at 25 and 20°C for C. perfringens . About 222 kinetics in foods were generated. The results were compared to simulations generated by existing software, such as PMP. The bias factor was also calculated. The methodology to obtain a food-dependent parameter (fitting step) and therefore to compare results given by models with new independent data (validation step) is discussed in regard to its food safety application. The proposed methods were used within the French national program of predictive microbiology, Sym′Previus, to include challenge test results in the database and to obtain predictive models designed for microbial growth in food products.

Lactic acid bacteria were selected for their inhibitory activity against Clostridium tyrobutyricum under conditions that eliminate the effects of lactic acid and hydrogen peroxide. Four strains were isolated belonging to the species Lactococcus lactis ssp. lactis. The sensitivity of the inhibitory substances to pronase and trypsine indicates that they are proteins or peptides different from nisin. Their resistance to phospholipase D indicates that they are also different from lactostrepcin. The inhibitory substances are produced during the exponential phase of growth. Their activity is bactericidal and directed toward some strains of Clostridium tyrobutyricum, Lactobacillus helveticus, and Streptococcus thermophilus, but strains used as dairy starters, Lactobacillus lactis, Streptococcus thermophilus, and Propionibacterium shermanii, are not all affected by the inhibition.