Influence of substrate and low temperature on growth and survival of verotoxigenicEscherichia coli

Growth and survival of verotoxigenicEscherichia coli(VTEC) were studied in tryptic soy broth (TSB), brain–heart infusion (BHI), and whole milk at 12, 9.5, 8.5, 7.5, 6.5, and 5.5°C (±0.2°C). Colony forming units (cfu) were enumerated by surface plating on eosin methylene blue (EMB) agar plates and incubating at 37°C for 24 h. Growth curves were plotted and lag and generation times were determined. Results indicated that the average generation times for each strain was similar in each medium (4.6±0.9 h) at 12°C. However, at 9.5°C the average generation time differed significantly among the media (P<0.05). The average generation time was longer in BHI compared to TSB and whole milk at 9.5°C (14 h in BHI vs 11 h in milk and TSB). Also, the generation time of individual strains in BHI varied from 9.5–22 h at 9.5°C. Decrease of temperature to 8.5°C resulted in lack of growth of two strains in BHI. At 7.5°C none of the strains grew in BHI, one grew in TSB, whereas all grew in milk. A further decrease of temperature to 6.5°C did not allow growth of any of the strains in TSB or BHI whereas all grew in milk. Addition of 5% lactose to TSB and BHI enabled survival and growth of all strains at 6.5°C. The results of this study demonstrate the influence of growth medium composition on the minimum growth temperature of verotoxigenicE. coli.     

Effect of Temperature, Solute and pH on the Tolerance of Clostridium perfringens to Reduced Water Activities

Two strains of C. perfringens type A (FDI and S45) were grown in Thioglycolate medium adjusted to a_ws of 0.995, 0.975, or 0.965 by the addition of NaCl, KCL or LiCl. Combinations of controlled and uncontrolled pHs (7.0, 6.5 and 6.0) and incubation temperatures of 45°C, 37°C, or 30°C were observed at each a_w. Maximal numbers of cells and shortest lag times occurred at a_w of 0.995, 45°C and pH 7.0. No growth occurred at a_w of 0.965. At 0.985, growth did not occur when KCl was used as the solute and the temperature and pH were 30°C and 6.0, respectively. NaCl was less inhibiting than KCl. LiCl inhibited growth completely.     

The growth and aflatoxin production of Aspergillus flavus strains on a cheese model system are influenced by physicochemical factors

Traditional cheeses may be contaminated by aflatoxin-producing Aspergillus flavus during the ripening process, which has not been sufficiently taken into account. The objectives of this study were to evaluate the influence of water activity (a_w), pH, and temperature on the lag phases, growth, and aflatoxin production of 3 A. flavus strains (CQ7, CQ8, and CG103) on a cheese-based medium. The results showed that the behavior of A. flavus strains was influenced by pH, a_w, and temperature conditions. The CQ7 strain showed the maximum growth at pH 5.5, 0.99 a_w, and 25°C, whereas for CQ8 and CQ103 strains, no differences were obtained at pH 5.5 and 6.0. In general, low pH, a_w, and temperature values increased the latency times and decreased the growth rate and colony diameter, although a_w and temperature were the most limiting factors. Maximum aflatoxin production on the cheese-based medium occurred at pH 5.0, 0.95 a_w, and 25 or 30°C, depending on the strain. This study shows the effect of pH, a_w, and temperature factors on growth and aflatoxin production of 3 aflatoxigenic A. flavus strains on a cheese-based medium. The findings may help to design control strategies during the cheesemaking process and storage, to prevent and avoid aflatoxin contamination by aflatoxigenic molds.     

Effect of pre-incubation conditions on growth and survival of Staphylococcus aureus in sliced cooked chicken breast

In this work, the effect of pre-incubation conditions (temperature: 10, 15, 37 °C; pH 5.5, 6.5 and water activity, a_w: 0.997, 0.960) was evaluated on the subsequent growth, survival and enterotoxin production (SE) of Staphylococcus aureus in cooked chicken breast incubated at 10 and 20 °C. Results showed the ability of S. aureus to survive at 10 °C when pre-incubated at low a_w (0.960) what could constitute a food risk if osmotic stressed cells of S. aureus which form biofilms survive on dried surfaces, and they are transferred to cooked meat products by cross-contamination. Regarding growth at 20 °C, cells pre-incubated at pH 5.5 and a_w 0.960 had a longer lag phase and a slower maximum growth rate. On the contrary, it was highlighted that pre-incubation at optimal conditions (37 °C/pH 6.5/a_w 0.997) produced a better adaptation and a faster growth in meat products what would lead to a higher SE production. These findings can support the adoption of management strategies and preventive measures in food industries leading to avoid growth and SE production in meat products.     

Characteristics of lactobacilli isolated from radurised meat

The properties of 67 lactic acid bacteria, isolated from radurised minced beef, were investigated. Only two of these strains were found to be hetero-fermentative and were identified as Lactobacillus viridescens and L. brevis. With one exception (Streptococcus lactis), all the strains belonged to the subgenus Streptobacterium, and represented L. saké, L. curvatus, L. bavarius, L. plantarum, L. alimentarius and L. faciminis. However, 7 strains were unidentifiable to species level according to traditional criteria. Several others were atypical in their physiological behaviour. Radiation sensitivity ranged from strains that had a D₁₀ value of 0.28 kGy to 0.88 kGy for the most resistant strain. No correlation was observed between H₂O₂ production and sensitivity to H₂O₂ or ionising radiation. Most strains produced inhibitory substances, seemingly bacteriocinogenic. An examination of technologically important characteristics revealed proteolytic activity for the majority of strains, whereas none of the strains were lipolytic. The lowest water activity that supported growth was a_w = 0.94. All strains had a psychrotrophic nature and grew at 4°C in MRS broth. Electron microscopial observations suggested a possible relationship between cell diameter and radiation resistance. These characteristics, together with their facultative aerobic nature and their relatively high resistance towards irradiation, may explain the dominance of lactobacilli in radurised, vacuum-packaged meat.     

Modeling of germination and growth of ochratoxigenic isolates of Aspergillus ochraceus as affected by water activity and temperature on a barley-based medium

Aspergillus ochraceus is an ochratoxin producing fungus that can be found on stored cereal grains such as barley. The objective of this study was to determine the effects of water activity (a_w, 0.75–0.99) and temperature (10–30°C) on germination and growth on barley extract agar medium (BMEA) of three isolates of Aspergillus ochraceus. The three isolates showed an optimal a_w for germination and growth of 0.99–0.95 at 20–30°C, with a marked increase of the lag phases and decrease of germination and growth rates at the marginal levels of a_w and temperature assayed. Minimum level of a_w for germination was 0.80 and 0.85 for growth. Data were then modeled by an MLR regression and response surface models were obtained. These models may allow a rough prediction of germination/growth as a function of the storage temperature and moisture content of barley grains.     

The effect of aeration,initial inoculum and meat microflora on the growth kinetics of Listeria monocytogenes in selective enrichment broths

The effect of increased aeration, initial inoculum and the meat microflora on the growth kinetics of Listeria monocytogenes in meat samples enriched in Listeria selective broth were investigated. Growth rate and lag phase duration were calculated following linear regression analysis on the growth data. Neither aeration or variations in the initial inoculum levels altered the length of the lag phase or the growth rate of L.monocytogenes. The growth kinetics of the meat microflora was similar under all of the experimental conditions investigated. Overall the meat microflora was shown to have a faster growth rate than L. monocytogenes in selective media. Organisms growing during Listeria selective enrichment were identified as Lactobacillus plantarum, Lactobacillus delbrueckii and pseudomonads. The implications of the growth of these organisms in Listeria selective broths are discussed.     

Combined Effect of Water Activity, pH and Sub-optimal Temperature on Growth and Enterotoxin Production of Staphylococcus aureus

Staphylococcus aureus growth and enterotoxin production in relation to a, (0.99–0.87), pH (4.0–7.0) and temperature (8–30°C) was determined in Brain-heart-infusion broth with respectively NaCl and sucrose as humectants. Growth of S. aureus was not observed at a_w 0.85, at pH 4.3 or at 8°C. At 12°C no growth occurred at a, 0.90 or at a, 0.93 in combination with pH < 5.5. At a_w 0.96 no growth occurred at pH ≤ 4.9. Production of staphylococcal enterotoxin type A (SEA) occurred under nearly all conditions allowing growth. Production of SEB appears to be determined by a_w; at a_w 0.96 SEB was produced at all temperatures allowing growth, while at a, 0.93 SEB was hardly produced. Production of SEC and SEF was affected both by a_w and temperaure. Production of these toxins was rarely observed at a, 0.93. No SEC and SEF were produced at 12°C. Extrapolation of these findings to food is discussed.     

Mathematical models to predict kinetic behavior and aflatoxin production of Aspergillus flavus under various temperature and water activity conditions

Mathematical models were developed to predict fungal growth and aflatoxin production of Aspergillus flavus. Fungal growth and aflatoxin concentrations were measured. The Baranyi model was fitted to fungal growth and toxin production data to calculate kinetic parameters. Quadratic polynomial and Gaussian models were then fitted to μ_max and LPD (lag phase duration) values. The ranges of temperature and a _w values showing a μ_max value increase were 15–35°C and 0.891–0.984, respectively. LPD was only observed when the temperature was 20–35°C with a _w=0.891?0.972. The μ_{max growth} value increased up to 35°C with \(b_w = 0.2\left( {b_w = \sqrt {1 - a_w } } \right)\) , then values declined. LPD_growth values increased as the b _w value increased. The μ_max value for aflatoxins increased up to 25°C, but decreased after 30°C, indicating that the developed models are useful for describing the kinetic behavior of Aspergillus flavus growth and aflatoxin production.     

Antibacterial Effects and Cell Morphological Changes in Sfaphylococcus aureus Subjected to Low Ethanol Concentrations

Ethanol in low concentrations (5–10% weight) affected aerobic growth (at 37°C) of two strains of Stuphylococcus aureus. Growth inhibition in laboratory media was observed with ethanol about 6–7% weight water activities (a_w) of ～0.97. This suggested that the ethanol antibacterial effect was not only from its a_w lowering ability. Electron microscopy observations showed intense morphological modifications in cells in broth with 5% to 6.5% ethanol, and clearly indicated that cell wall changes of S. aureus was partly responsible for the antibacterial action of ethanol.     

Desorption isotherms of salted minced pork using K-lactate as a substitute for NaCl

The aim of this study was to obtain and compare water desorption isotherms of ground meat containing NaCl (0.107 kg NaCl/kg raw-meat dry matter) and/or K-lactate as NaCl substitute at two different levels of molar substitution (30% and 100%). A thin layer of salted ground meat was dried and sampled at pre-determined times. The moisture content of the samples and their water activities (a_w) were measured at 5 °C and 25 °C. Results showed that ground meat with NaCl and/or different K-lactate contents had a similar water desorption isotherm for a_w ranging from 0.7 to 1.0. Below 0.7, the water equilibrium content fell with small decreases in a_w faster for meat with NaCl than for meat with K-lactate. K-lactate could reduce the excessive hardening at the surface of salted meat products. Experimental desorption isotherms were compared to those estimated using two approaches of the Ross equation. Models provided a good fit for the experimental data.     

A Study of Factors Affecting Growth and Recovery of Listeria monocytogenes Scott A

The growth parameters for Listeria monocytogenes Scott A at various temperatures, pH values, water activity values, and in different carbohydrates were assessed. Growth was seen from 4°C through 45°C and at pH values from 4.7 through 9.2. Generation times were calculated for each parameter at which growth was observed. Optimum growth was defined as shortest generation time. L. monocytogenes grew optimally over the temperature range 30–37°C. The optimum pH was 7.0. Growth was demonstrated in solutions of up to 39.4% sucrose (a_w of 0.92). Carbohydrate fermentation was variable. Further studies involved examination of growth at optimal water activities (0.97) with suboptimal temperatures and pH values, and at 7.0, with suboptimal temperatures and water activities. Growth was seen when these parameters were combined.     

Influence of water activity and temperature on conidial germination and mycelial growth of ochratoxigenic isolates of Aspergillus ochraceus on grape juice synthetic medium. Predictive models

The first stages in the development of Aspergillus ochraceus, an ochratoxin A‐producing fungus that infects grapes and may grow on them, have been studied on a synthetic nutrient medium similar to grape in composition. Spore germination and mycelial growth have been tested over a water activity (a_w) and temperature range which could approximate to the real conditions of fungal development on grapes. Optimal germination and growth were observed at 30 °C for all three isolates tested. Maximal germination rates were detected at 0.96–0.99 a_w at 20 °C, while at 10 and 30 °C the germination rates were significantly higher at 0.99 a_w. Although this abiotic factor (a_w) had no significant influence on mycelial growth, growth rates obtained at 0.98 a_w were slight higher than those at other a_w levels. Predictive models for the lag phase before spore germination as a function of water activity and temperature have been obtained by polynomial multiple linear regression, and the resulting response surface models have been plotted. Copyright © 2005 Society of Chemical Industry     

Growth of Aeromonas hydrophila in modified-atmosphere-packed cooked meat products

Growth of Aeromonas hydrophila was investigated in modified-atmosphere-packed cooked meat products by developing predictive models. Modified brain–heart infusion (BHI) was shown to be suitable as a simulation medium for cooked meat products. Predictive models were developed for the growth parameters (maximum specific growth rate and lag phase) of A. hydrophila in modified BHI as a function of temperature, water activity and concentration of dissolved carbon dioxide. The growth of A. hydrophila was compared with the growth of the Specific Spoilage Organism (SSO) for cooked meat products Lactobacillus sake, to determine possible risk areas for A. hydrophila in modified atmosphere-packed cooked meat products. Aeromonas hydrophila was shown to multiply very rapidly at refrigerated temperatures. The developed models clearly demonstrated however that proliferation of A. hydrophila could be prevented by the use of carbon dioxide in the package atmosphere in combination with a decreased water activity (<0·985). Gas-packed cured cooked meat products will not sustain the growth of A. hydrophila when kept at refrigerated temperatures (<7°C).     

Comparisons of water activity and temperature impacts on growth of Fusarium langsethiae strains from northern Europe on oat-based media

This study has examined the effect of water activity (a_w, 0.995-0.90) and temperature (10-37 °C) on the lag phases prior to growth, growth rates and used models to develop two dimensional profiles for optimum and marginal conditions for two strains of Fusarium langsethiae from four northern European countries (UK, Norway, Sweden, and Finland) on an oat-based medium. Results showed that the optimum a_w for growth was at 0.98-0.995 and 25 °C. The limit for growth of the strains was at 0.92-0.93 a_w with minima of 10 °C. No growth occurred at 37 °C. The lag phases prior to growth were lowest under optimum conditions and extended to > 10 days at marginal conditions. Statistical analyses of intra and inter-strain differences in terms of both lag phases prior to growth and growth rates were not statistically significant. However, a_w and temperature were statistically significant factors. Two dimensional profiles for strains from each country of origin were built to identify optimum and marginal conditions for F. langsethiae for the first time. These environmental profiles will be beneficial for improving the ecological knowledge of this species which is able to produce trichothecene mycotoxins in a range of temperate cereals.     

Influence of water activity on microbial development in minced,cured pork,in relation to gas headspace composition

The water activity (a_w) of a cured minced meat mixture was reduced with NaCl or glycerol + NaCl and the products stored in air, N₂ or CO₂ at 4°C. The lower the a_w, the lower was the growth rate of the total microflora except when a_w was reduced below 0·98 with glycerol and products were stored in CO₂, when the growth rate was independent of a_w. At a_w 0·94 NaCl was more effective than glycerol + NaCl in reducing the growth rate. In air the final microflora of products stored at a_w ≤0·96 consisted primarily of yeasts while at 0.98 a_w, both Lactobacillus (62%) and yeasts (38%) were found. In CO₂ and N₂Lactobacillus predominated at 0·94–0·98 a_w when 2% NaCl or NaCl + glycerol were used. At 0·94 a_w, produced using 6% NaCl, a heterogenous final microflora was found, including Micrococcus and Staphylococcus. An increase in the microbial shelf-life of meat products may be obtained by reducing the a_w, particularly for use with storage in N₂- and CO₂-atmospheres. The type of solute chosen to regulate a_w may affect the solubility of CO₂, which in turn affects the shelf-life.     

Application of polynomial models to predict growth of mixed cultures of Pseudomonas spp. and Listeria in meat

Three models for one rapid and one slow growing strain of Pseudomonas fragi and one slow growing strain of P. fluorescens were developed in a meat broth; they were designed to take account of variations in growth and to provide a growth response interval. These models, and another for Listeria monocytogenes (Lm14 model), were used to predict the growth of spoilage Pseudomonas spp. and pathogenic Listeria in meat products. The Pseudomonas and Listeria models provided satisfactory predictions concerning inoculated strains grown in decontaminated beef meat. It was also possible to use the Pseudomonas models to predict the growth of the natural flora (mainly Pseudomonas spp.) of refrigerated meat stored under aerobic conditions. In experiments with mixed populations, three situations were observed: (1) in decontaminated meat, L. monocytogenes inoculated alone grew well at 6 degrees C, and this result was correctly predicted by the model; (2) in decontaminated meat inoculated with Listeria and Pseudomonas strains, L. innocua grew well and was not affected by the presence of Pseudomonas, and the growth of both organisms was correctly predicted by the models; (3) in naturally contaminated meat inoculated with Listeria, the strain did not grow until Pseudomonas had reached the stationary phase. The models satisfactorily predicted the growth of Pseudomonas spp. but not that of Listeria. In conclusion, the Lm14 model cannot be used for refrigerated meat stored aerobically as the results suggest a 'fail-safe' level which may be too high: meat had already reached a spoilage state even though no increase in the level of Listeria was observed. The Pseudomonas models accurately predicted the growth of naturally occurring Pseudomonas spp.     

Growth of Aeromonas hydrophila K144 as Affected by Organic Acids

The influence of different acids on the aerobic growth kinetics of Aeromonas hydrophila was studied in BHI broth with 0.5 and 2.0% NaCl incubated at 5 and 19°C. Growth curve data were analyzed by the Gompertz equation and a nonlinear regression program; generation and lag times were calculated from the Gompertz parameters. Type of acid, pH, NaCl level and temperature influenced lag and generation times. The organic acids (acetic, lactic, citric and tartaric) inhibited growth at higher pH values than inorganic acids (HCl and H₂SO₄). The high NaCl level interacted with type of acid and pH to restrict growth of the organism at the lower temperature of incubation. Acetic and lactic acids were effective in controlling the growth of A. hydrophila and could readily be combined with low holding temperature to render foods free of the organism.     

Modeling Penicillium Expansum Growth Response to Thyme Essential oil at Selected Water Activities and pH Values Using Surface Response Methodology

Our objective was to evaluate, using a full factorial design, the effects of selected water activities (0.990, 0.945, or 0.900), pHs (5, 4, or 3), and thyme essential concentration (TEO, 0, 25, 50, or 100ppm) oil on Penicillium expansum lag time (λ) and radial growth rate (μ_m) obtained by modeling mold response using Gompertz equation, and corresponding polynomial quadratic models. Potato-dextrose agar formulated with every studied factor combination was inoculated with 10³ spores/ml, and incubated at 25°C up to 30 days. Mold colony diameter was periodically measured during incubation and adjusted with Gompertz equation to determine λ and μ_m. Decreasing a_w and pH, and increasing TEO concentration decreased μ_m and increased λ. At low a_w and pH, the increase in TEO concentration had a dramatic effect on P. expansum response since 25ppm of TEO inhibited its growth for 30 days at 25°C. Gompertz parameters exhibited that P. expansum was sensitive to the evaluated combined factors, allowing us to construct a secondary predictive growth model. TEO in combination with a_w and pH reduction effectively inhibited P. expansum growth.     

Influence of different histories of the inoculum on lag phase and growth of Listeria monocytogenes in meat models

The aim of this study was to determine the effect of history of inoculum and preservatives on the lag phase and growth rate of Listeria monocytogenes strains in meat products packaged under modified atmosphere conditions. Inocula with different histories were added to meat models, and growth rate and lag phase of two strains of L. monocytogenes were measured at 5 and 10 degrees C. The meat model stored at 10 degrees C contained sodium lactate, but the model stored at 5 degrees C did not. The five different histories of the inocula included cold propagation, biofilm formation, and starvation. The lag phase ranged from 1 to 10 days and was affected by the history of the inoculum, whereas the growth rate was constant except for one combination of history of inoculum and strain, where growth did not start during the incubation period. In a second series of experiments, the growth rate and lag phase of the two Listeria strains and the effects of two different histories of inoculum were tested in meat models with pH 5.7 or 6.5 and increasing amounts of NaCl. The growth rate depended on salt concentration, bacterial strain, and pH, whereas lag phase duration depended on history of inoculum, salt concentration, and pH. The lag phase duration was highly dependent on the history of the inoculum, and higher amounts of preservative (NaCl) made these effects even more noticeable. The results of this study underline the importance of the effects of the history of the inoculum on lag phase duration and could be used to predict lag phase in industrial meat products.