A stochastic approach for integrating strain variability in modeling Salmonella enterica growth as a function of pH and water activity

Strain variability of the growth behavior of foodborne pathogens has been acknowledged as an important issue in food safety management. A stochastic model providing predictions of the maximum specific growth rate (μ_max) of Salmonella enterica as a function of pH and water activity (a_w) and integrating intra-species variability data was developed. For this purpose, growth kinetic data of 60 S. enterica isolates, generated during monitoring of growth in tryptone soy broth of different pH (4.0-7.0) and a_w (0.964-0.992) values, were used. The effects of the environmental parameters on μ_max were modeled for each tested S. enterica strain using cardinal type and gamma concept models for pH and a_w, respectively. A multiplicative without interaction-type model, combining the models for pH and a_w, was used to describe the combined effect of these two environmental parameters on μ_max. The strain variability of the growth behavior of S. enterica was incorporated in the modeling procedure by using the cumulative probability distributions of the values of pH_min, pH_opt and a_wmin as inputs to the growth model. The cumulative probability distribution of the observed μ_max values corresponding to growth at pH 7.0-a_w 0.992 was introduced in the place of the model's parameter μ_opt. The introduction of the above distributions into the growth model resulted, using Monte Carlo simulation, in a stochastic model with its predictions being distributions of μ_max values characterizing the strain variability. The developed model was further validated using independent growth kinetic data (μ_max values) generated for the 60 strains of the pathogen at pH 5.0-a_w 0.977, and exhibited a satisfactory performance. The mean, standard deviation, and the 5th and 95th percentiles of the predicted μ_max distribution were 0.83, 0.08, and 0.69 and 0.96 h^− 1, respectively, while the corresponding values of the observed distribution were 0.73, 0.09, and 0.61 and 0.85 h^− 1. The stochastic modeling approach developed in this study can be useful in describing and integrating the strain variability of S. enterica growth kinetic behavior in quantitative microbiology and microbial risk assessment.     

On the selection of relevant environmental factors to predict microbial dynamics in solidified media

Several studies have shown that food structure causes slower growth rates and narrower growth boundaries of bacteria compared to laboratory media. In predictive microbiology, both a_w or corresponding solute concentration (mainly NaCl) have been used as a growth influencing factor for kinetic models or growth/no growth interface models. The majority of these models have been based on data generated in liquid broth media with NaCl as the predominant a_w influencing solute. However, in complex food systems, other a_w influencing components might be present, next to NaCl. In this study, the growth rate of Salmonella typhimurium was studied in the growth region and the growth/no growth response was tested in Tryptic Soy Broth at 20 °C at varying gelatin concentration (0, 10, 50 g L⁻¹ gelatin), pH (3.25–5.5) and water activity (a_w) (0.929–0.996). From the viewpoint of water activity, the results suggest that NaCl is the main a_w affecting compound. However, gelatin seemed to have an effect on medium a_w too. Moreover, there is also an interaction effect between NaCl and gelatin. From the microbial viewpoint, the results confirmed that the a_w decreasing effect of gelatin is less harmful to cells than the effect of Na⁺ ions. The unexpected shift of the growth/no growth interface to more severe conditions when going from a liquid medium to a medium with 10 g L⁻¹ gelatin is more pronounced when formulating the models in terms of a_w than in terms of NaCl concentrations. At 50 g L⁻¹ gelatin, the model factored with NaCl concentration shifts to milder conditions (concordant to literature results) while the model with a_w indicates a further shift to more severe conditions, which is due to the water activity lowering effect of gelatin and the interaction between gelatin and NaCl. The results suggest that solute concentration should be used instead of a_w, both for kinetic models in the growth region and for growth/no growth interface models, if the transferability of models to solid foods is to be increased.     

Modelling the effect of temperature and water activity on the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea

The purpose of the present study was to apply a modelling approach to define the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea on a synthetic medium as a function of temperature and water activity. Both fungal species were grown on malt extract agar at different temperatures (10, 15, 20, 25, 30, 35, 40 and 45 °C) and a_w levels (0.88, 0.90, 0.92, 0.94, 0.96 and 0.99) for a period of 30 days. Growth responses were evaluated over time in terms of colony diameter changes. Growth data were fitted to the primary model of Baranyi and the resulting growth rates were further modeled as a function of temperature and water activity using the cardinal model with inflection (CMI) ( Rosso et al., 1993). A logistic regression quadratic polynomial model was also employed to predict the probability of growth over storage time. Estimated parameters for minimum, maximum and optimum temperatures for growth were 9.1 °C, 46.4 °C and 32.1 °C for B. fulva and 10.5 °C, 43.2 °C and 32.1 °C for B. nivea. The respective values for a_w were 0.893, 0.993 and 0.985 for B. fulva and 0.892, 0.992 and 0.984 for B. nivea. No growth was observed at 0.88 a_w regardless of temperature for both species, whereas B. nivea ascospores could not grow at 10 and 45 °C irrespective of a_w. Regarding growth boundaries, the degree of agreement between predictions and observations was >98% concordant for both species. The erroneously predicted growth cases were 1.4–4.2% false positive and 2.1–3.5% false negative for B. nivea and B. fulva, respectively. The developed logistic model was validated with two literature data sets as well as with data from independent experiments carried out on fruit juices. Validation results showed that agreement with literature data for growth was 25 out of 36 (69.4%) cases, whereas validation on fruit juice data failed in only 6 cases (5 false positives and 1 false negative) out of 128 cases.     

Evaluation of ability of ferulic acid to control growth and fumonisin production of Fusarium verticillioides and Fusarium proliferatum on maize based media

The aim of this study was to evaluate the efficacy of ferulic acid (1, 10, 20 and 25 mM) at different water activity (a_w) values (0.99, 0.98, 0.96 and 0.93) at 25 °C on growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on maize based media. For both Fusarium species, the lag phase significantly decreased (p ≤ 0.001), and the growth rates increased (p ≤ 0.001) at the lowest ferulic acid concentration used (1 mM), regardless of the a_w. However, high doses of ferulic acid (10 to 25 mM) significantly reduced (p ≤ 0.001) the growth rate of both Fusarium species, regardless of the a_w. In general, growth rate inhibition increased as ferulic acid doses increased and as media a_w decreased. Fumonisin production profiles of both Fusarium species showed that low ferulic acid concentrations (1–10 mM) significantly increased (p ≤ 0.001) toxin production, regardless of the a_w. High doses of ferulic acid (20–25 mM) reduced fumonisin production, in comparison with the controls, by both Fusarium species but they were not statistically significant in most cases. The results show that the use of ferulic acid as a post-harvest strategy to reduce mycotoxin accumulation on maize needs to be discussed.     

Rheological evaluation of gelatin–xanthan gum system with high levels of co-solutes in the rubber-to-glass transition region

Effects of moisture content, xanthan gum (XG) addition, and glucose syrup (GS):sucrose ratio on the gelation of gelatin-XG systems with high levels of co-solutes were investigated in the rubbery and the glass transition regions. Frequency sweep tests were performed between 0.1 and 100 rad and the storage (G′) and loss (G″) moduli of the system were measured in the temperature range of 60 to −15 °C. The onset of glass transition region increased with decreasing moisture content. The time–temperature superposition yielded master curves of G′ and G″ as a function of timescale of measurement. G″ and G″ were superimposed with the horizontal shift factor a_T, which was temperature dependent according to the Williams–Landel–Ferry (WLF) equation. Glass transition temperature (T_g) of the samples were determined by dynamic mechanical analysis (DMA) from the peak of tan δ. T_g decreased with XG addition. The energy of vitrification of samples with XG increased compared to samples containing only gelatin. Relaxation spectra of the samples were calculated from rheological measurements using the first and second approximations. The Rouse theory was more closely followed with the second approximation.     

Influence of water activity and temperature on growth and mycotoxin production by Alternaria alternata on irradiated soya beans

The aim of this study was to determine the effects of water activity (a_w) (0.99-0.90), temperature (15, 25 and 30 °C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 a_w and 25 °C. Minimum a_w level for growth was dependent on temperature. Both strains were able to grow at the lowest a_w assayed (0.90). Maximum amount of AOH was produced at 0.98 a_w but at different temperatures, 15 and 25 °C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 a_w and 30 °C for both strains. The concentration range of both toxins varied considerably depending on a_w and temperature interactions. The two metabolites were produced over the temperature range 15 to 30 °C and a_w range 0.99 to 0.96. The limiting a_w for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of a_w × temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and a_w conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.     

Effect of water activity, temperature and incubation time on growth and ochratoxin A production by Aspergillus niger and Aspergillus carbonarius on maize kernels

The aim of this study was to determine the effects of water activity (a_w) (0.92-0.98), temperature (5-45 °C) and incubation time (5-60 days) on growth and ochratoxin A (OTA) production by Aspergillus niger and Aspergillus carbonarius on maize kernels using a simple method. Colony diameters of both strains at 0.92 a_w were significantly lower than those at 0.96 and 0.98 a_w levels. The optimum growth temperature range for A. niger was 25-40 °C and for A. carbonarius 20-35 °C. A. niger produced OTA from 15 to 40 °C, and the highest OTA level was recorded at 15 °C. The concentration of OTA produced at 0.92 a_w was significantly lower than those at 0.96 and 0.98 a_w. A. carbonarius produced OTA from 15 to 35 °C and the maximum concentration was achieved at 15 °C, although not differing statistically from the concentration detected at 20 °C. At 0.98 a_w the OTA concentration was significantly higher than at 0.96 and 0.92 a_w. Our results show that maize supports both growth and OTA production by A. niger and A. carbonarius. The studied strains were able to produce OTA in maize kernels from the fifth day of incubation over a wide range of temperatures and water availabilities. Although the limit of quantification of our method was higher than that required for the analysis of OTA in food commodities, it has proved to be a useful and rapid way to detect OTA production by fungi inoculated onto natural substrates, in a similar way as for pure culture. Both species could be a source of OTA in this cereal in temperate and tropical zones of the world.     

Growth and survival of Salmonella in ground black pepper (Piper nigrum)

A four serovar cocktail of Salmonella was inoculated into ground black pepper (Piper nigrum) at different water activity (a_w) levels at a starting level of 4–5 log cfu/g and incubated at 25 and at 35 °C. At 35 °C and a_w of 0.9886 ± 0.0006, the generation time in ground black pepper was 31 ± 3 min with a lag time of 4 ± 1 h. Growth at 25 °C had a longer lag, but generation time was not statistically different from growth at 35 °C. The a_w threshold for growth was determined to be 0.9793 ± 0.0027 at 35 °C. To determine survival during storage conditions, ground black pepper was inoculated at approximately 8 log cfu/g and stored at 25 and 35 °C at high (97% RH) and ambient (≤40% RH) humidity. At high relative humidity, a_w increased to approximately 0.8–0.9 after approximately 20 days at both temperatures and no Salmonella was detected after 100 and 45 days at 25 and 35 °C, respectively. Under ambient humidity, populations showed an initial decrease of 3–4 log cfu/g, then remained stable for over 8 months at 25 and 35 °C. Results of this study indicate Salmonella can readily grow at permissive a_w in ground black pepper and may persist for an extended period of time under typical storage conditions.     

Mechanical spectra and calorimetric evaluation of gelatin–xanthan gum systems with high levels of co-solutes in the glassy state

Mechanical properties of gelatin–xanthan gum (XG) mixtures with high levels of co-solutes were examined by dynamic mechanical analysis (DMA). The mechanical spectra of the samples were modeled according to the Williams–Landel–Ferry (WLF) equation/free-volume theory, which requires an entropic lightly cross-linked network. For the α dispersion, E′ and E′′ superposed with the horizontal shift factor a_T, which was temperature-dependent according to the WLF equation; no other secondary dispersion mechanism was detected. The addition of XG to gelatin networks with high levels of co-solutes changed the glass transition temperature (T_g) and kinetics of glass transition and glassy states. In the glassy state, the WLF equation was unable to follow progress in the mechanical properties, which were better described by the Andrade equation. The calorimetric measurements of the gelatin–XG systems were made using a modulated temperature differential scanning calorimetry (MTDSC) to improve the determination of T_g. The samples were exposed to two cooling and heating cycles to provide a controlled recent thermal history in the temperature range of 40 °C to −70 °C. The T_g values of the samples were determined from the second heating cycle in the reversing heat signal. The calorimetric T_g values increased with increasing glucose syrup:sucrose ratio due to increased crosslinking, whereas mechanical T_g decreased with increased XG content due to network formation.     

Development and application of predictive models for fungal growth as tools to improve quality control in yogurt production

The effect of storage temperature (0–40 °C) and inoculum size (10¹–10⁵ spores) on the mycelium growth kinetics of 12 fungal species on yogurt were monitored. A cardinal model with inflection (CMI) was used to describe the effect of temperature on the growth rate (μ) and the lag time (λ) of each isolate. Significant differences on the temperature dependence of the mycelium growth between the tested species were observed. Depending on the strain, the estimated minimum, optimum and maximum temperature parameters for μ (T_min, T_opt, T_max) ranged from −7.6 to 9.6, 19.5 to 37.8 and 29.8 to 46.9 °C, respectively. Only λ was found to be affected by the inoculum size and a linear relation between Ln (λ) and Log (inoculum size) was revealed. The inoculum level did not influence the values of T_min, T_opt and T_max for λ. Based on the above observations, the combined effect of inoculum size and temperature on λ was modeled using a modified CMI. The parameter λ_opt (λ at optimum conditions) was expressed as a function of the inoculum size. Validation studies showed a good performance of the developed models. The application scheme of the models for improving fungi control in yogurt productions is discussed.     

Effect of antioxidants and competing mycoflora on Fusarium verticillioides and F. proliferatum populations and fumonisin production on maize grain

This study was carried out to determine the temporal effect of the antioxidants butylated hydroxyanisol (BHA) and propyl paraben (PP) at doses of 500 and 1000 μg/g on the growth of Fusarium verticillioides and F. proliferatum inoculated on natural maize grain in the presence of the competing mycoflora and fumonisin production at 0.98 and 0.95 water activity (a_w) over a 28-day storage period. The reduction in the log colony forming units (CFU) of Penicillium, Aspergillus and Fusarium populations was 10-100 fold depending on dose of BHA or PP, a_w and time. However, the populations of all three groups were higher at 0.98 a_w than 0.95 a_w. BHA at 500 μg/g and 0.95 a_w reduced the fumonisin content by 82% after 7-14 days incubation, but at the end of the experimental period the reduction was only 32%. A higher reduction in the level of fumonisin produced (77%) was achieved with BHA at 1000 μg/g after 28 days. PP at 500 and 1000 μg/g decreased fumonisin production throughout the incubation period in the drier treatment, but at 0.98 a_w control of toxin production was only achieved after 7-14 days. The reduction in the fumonisin levels could be due to the combined effect of antioxidants, and the competing mycoflora, mainly Aspergillus and Penicillium species.     

Influence of temperature, pH and NaCl concentration on the maximal growth rate of Brochothrix thermosphacta and a bioprotective bacteria Lactococcus piscium CNCM I-4031

The maximum specific growth rate (μ_max) of Brochothrix thermosphacta, a spoilage bacteria of cooked peeled shrimp, and Lactococcus piscium CNCM I-4031, a bioprotective strain, was investigated under different conditions of temperature, NaCl concentrations and pH. The basic modelling approach used was the Gamma concept (γ-concept) and the model developed was then adapted to shrimp. Cardinal growth parameters were quite similar for the two strains, except for NaCl. No NaCl was required for growth and the NaCl_max was three-times higher for B. thermosphacta than for L. piscium (62 and 23 g l⁻¹ respectively). However, tolerance to NaCl was higher in seafood than in liquid broth, possibly due to presence of osmoltically active molecules. L. piscium and B. thermosphacta were psychrotolerant, with T_min = −4.8 and −3.4 °C, T_opt = 23.4 and 27.0 °C and T_max = 27.2 and 30.8 °C respectively. The optimal pH was neutral and growth possible till pH = 4.8 for the two strains, assuming possible applications of the bioprotective strain in lightly marinated seafood. The μ_max of B. thermosphacta in shrimp was a little higher than in L. piscium whatever the environmental conditions. Validation of the model showed that the γ-concept was suitable for predicting μ_max of B. thermosphacta in shrimp. Data generated in this study can be used to adapt the model to other foods with few additional experiments and the effect of different parameters may be added in the future. The model was less accurate for the bioprotective strain and the effect of NaCl must be studied in more detail directly in the matrix.     

Development and application of a predictive model of Aspergillus candidus growth as a tool to improve shelf life of bakery products

Molds are responsible for spoilage of bakery products during storage. A modeling approach to predict the effect of water activity (a_w) and temperature on the appearance time of Aspergillus candidus was developed and validated on cakes. The gamma concept of Zwietering was adapted to model fungal growth, taking into account the impact of temperature and a_w. We hypothesized that the same model could be used to calculate the time for mycelium to become visible (t_v), by substituting the matrix parameter by t_v. Cardinal values of A. candidus were determined on potato dextrose agar, and predicted t_v were further validated by challenge-tests run on 51 pastries. Taking into account the a_w dynamics recorded in pastries during reasonable conditions of storage, high correlation was shown between predicted and observed t_v when the a_w at equilibrium (after 14 days of storage) was used for modeling (A_f = 1.072, B_f = 0.979). Validation studies on industrial cakes confirmed the experimental results and demonstrated the suitability of the model to predict t_v in food as a function of a_w and temperature.     

Water migration mechanisms in amorphous powder material and related agglomeration propensity

The agglomeration phenomenon of amorphous particulate material is a major problem in the food industry. Currently, the glass transition temperature (T_g) is used as a fundamental parameter to describe and control agglomeration. Models are available that describe the kinetics of the agglomeration process as a function of the distance of the material from T_g (i.e. T − T_g). In practice these models are often not applied because they assume that the powder material is instantly in equilibrium with the humidity conditions of the environment and that solid mobility only occurs at T > T_g. Insights in the kinetics and mechanisms of water transport in powder material can help to better understand and control powder agglomeration. For this purpose, gravimetric step-change water sorption experiments were performed on maltodextrins as a function of the water activity a_w. The maltodextrins vary in dextrose equivalents (DE), particle size and morphology. The experimental results were compared with a Fickian diffusion model in order to understand the dependency of the transport mechanism on water concentration gradient and material relaxation. The water transport kinetics in the maltodextrins with low DE (i.e. 6) were well described by Fickian diffusion for low a_w, independently of particle size and morphology, until relaxation phenomena started to occur at an a_w corresponding to T − T_g = −20 °C. The importance of the matrix relaxation phenomena on the water transport mechanism increased with increasing DE (i.e. 29 and 32), not showing any relationship with the T_g. The results of this study indicate that the water migration mechanism is controlled by relaxation phenomena when the amorphous material is still far from the glass–rubber transition. The T − T_g at which the relaxation phenomena occur depends on the material. On the contrary, the T − T_g parameter could well describe the onset of agglomeration, independently from the material properties. Therefore, it can be concluded that within the conditions of this study, matrix relaxation occurring far below T_g did not affect the onset of agglomeration.     

Water activity and temperature effects on mycotoxin production by Alternaria alternata on a synthetic tomato medium

Alternaria spp. have been reported to be the most frequent fungal species invading tomatoes. Certain species, in particular the most common one, A. alternata, are capable of producing several mycotoxins in infected plants and in agricultural commodities. Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TA) are some of the main Alternaria mycotoxins that can be found as contaminants of food. The objective of this study was to determine the effect of water activity (a_w, 0.904, 0.922, 0.954, and 0.982) and temperature (6, 15, 21 and 35 °C) on mycotoxin production on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The optimum AOH production occurred at 0.954 a_w after 28 days of incubation at 21 °C. A temperature of 21 °C was the most favourable for AOH synthesis at all a_w levels. The maximum concentration of AME was determined at 0.954 a_w and 35 °C. The optimum conditions for TA accumulation were 0.982 a_w and 21 °C. At the 0.904 a_w no growth or germination was registered at 6 °C and 15 °C over the whole incubation period. At 21 °C and 35 °C growth occurred slowly but none of the toxins were detected at this a_w level. In general, high a_w levels were favourable for mycotoxin production. None of the other toxins was detected at quantifiable levels at 6 °C after the whole incubation period. A storage temperature of 6 °C or below could be considered as safe for tomato fruits and high moisture tomato products (a_w > 0.95), in relation with Alternaria toxins. The results obtained here could be extrapolated to evaluate the risk of spoilage in tomato fruits and tomato products caused by this pathogen.     

Effect of fenpropimorph, prochloraz and tebuconazole on growth and production of T-2 and HT-2 toxins by Fusarium langsethiae in oat-based medium

Fusarium langsethiae has been isolated from infected cereals in central and northern Europe where it has been identified in the last decade as the main species involved in the occurrence of high levels of T-2 and HT-2 toxins, mainly in oats. The efficacy of three fungicides (prochloraz, tebuconazole, fenpropimorph) for controlling growth of two strains of F. langsethiae isolated from oats was examined at 0.96 and 0.98 a_w at 15, 20 and 25 °C on oat-based media. The concentrations necessary for 50 and 90% growth inhibition (ED₅₀ and ED₉₀ values) were determined. The effect on the trichothecene type A mycotoxins T-2 and HT-2 was also determined. Without fungicides both strains grew faster at 0.98 than at 0.96 a_w and the influence of temperature on growth rates was 25 > 20 > 15 °C. Prochloraz and tebuconazole were more effective than fenpropimorph against F. langsethiae. Strain, temperature and type of fungicide significantly influenced the ED₅₀ and ED₉₀ values for growth. The concentration ranges under different environmental conditions were: prochloraz (0.03-0.1 and 0.3-1.5), tebuconazole (0.06-0.9 and 1.3-8.2), and fenpropimorph (22-59 and 125-215 mg l⁻¹). Production of T-2 and HT-2 toxins was influenced by temperature, a_w, type of fungicide and dose. Levels of T-2 were usually higher than those of HT-2 under the same conditions. The biosynthesis of T-2 toxin increased after 10 day incubation, but was reduced with decreasing temperature and increasing fungicide dose. At 0.98 a_w T-2 levels increased in cultures containing fenpropimorph while at 0.96 a_w the toxin concentrations increased in response to the other two fungicides. Low doses of prochloraz or tebuconazole enhanced toxin production when compared with untreated cultures for strain 2004-59 at 0.96 a_w and 20-25 °C. HT-2 was hardly detectable in the treatments with prochloraz or tebuconazole at 0.98 a_w. This is the first study on the effect of these anti-fungal compounds on control of growth of F. langsethiae and on production of T-2 and HT-2 toxins in oat-based media.     

Effect of various salts and pH condition on rheological properties of Salvia macrosiphon hydrocolloid solutions

The extraction of Salvia macrosiphon seed hydrocolloid was performed successfully and the optimum hydration time and temperature were found to be 30 min and 25 °C for 12% extraction yield. The effect of solution pH (3–9) and addition of NaCl (0.5–3%), CaCl₂ (0.5–3%) and Na₂HPO₄ (0.2–0.6%) on rheological properties of the hydrocolloid solutions were investigated. Rheologcal data were fitted with the Power law model with regression coefficient of greater than 0.95. Strong shear thinning behavior with the n value of less than 0.53 was recorded. All variables had significant impacts on rheological parameters. The minimum and maximum consistencies were found at pH 3 and the 9, respectively. This was explained with the presence of carboxylic groups on the biopolymer backbone as indicated by FTIR. NaCl, CaCl₂ and Na₂HPO₄ had significant effects on rheological properties. Moreover, at similar salt concentration of 0.5%, Na₂HPO₄ had the greatest and NaCl had the lowest effect.     

Studying the growth boundary and subsequent time to growth of pathogenic Escherichia coli serotypes by turbidity measurements

The presence of Escherichia coli in contaminated food products is commonly attributed to faecal contamination when they are improperly handled and/or when inactivation treatments fail. Adaptation of E. coli at low pH and a_w levels can vary at different temperatures depending on the serotype, thus more detailed studies are needed. In this work, a screening to assess the growth of four pathogenic serotypes of E. coli (O55:H6; O59:H21; O158:H23 and O157:H7) was performed. Subsequently, boundary models were elaborated with the fastest serotype selected at different temperatures (8, 12 and 16 °C), and inoculum levels (2, 3 and 4 log cfu/mL) as function of pH (7.00–5.00) and a_w (0.999–0.960). Finally, the growth kinetics of E. coli was described in the conditions that allowed growth. Results obtained showed that the serotypes O157:H7 and O59:H21 did not grow at more stringent conditions (8 °C; pH 5.50), while the E. coli O158:H23 was the best adapted, resulting in faster growth. The logistic regression models presented a good adjustment to data observed since more than 96.7% of cases were correctly classified. The growth interface was shifted to more limited conditions as the inoculum size was higher. Detection times (t_d, h) and their variability were higher at low levels of the environmental factors studied. This work provides insight on the growth kinetics of E. coli at various environmental conditions.     

Application of Central Composite Design to evaluate the antilisterial activity of hydro-alcohol berry extract of Myrtus communis L.

The antimicrobial activity of the hydro-alcohol extract of Myrtus communis L. (ME) berries was investigated against six Listeria monocytogenes strains (2 type strains and 4 isolates). Sub-lethal ME concentrations reduced L. monocytogenes counts by at least 2 log cycles. A Central Composite Design was used to investigate the combined effects of sub-lethal concentrations of ME (0.039–0.195 mL/100 mL), NaCl (0–2.0 g/100 mL) and pH (5.0–7.0) on strains growth. ME affected growth parameters, generally extending lag phase length and reducing maximum growth, sometimes with interactive effects with pH. The highest ME concentrations (0.117–0.195 mL/100 mL) combined with the lowest pH values (5.0–6.0) strongly reduced or even inhibited strains growth.