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The article explains the development of a new experimental food elastic system that can be used for analyzing the yeast spoilage taking place in different acidic sauces. The model is shown to be extremely effective and efficient and can be easily applied to several different experiments.

In a gelatin-dextran mixture, changing the (relative and/or absolute) concentration of the components leads to the formation of different microstructures. Confocal laser scanning microscopy illustrated that the nature of the microstructure determines the location and morphology of Escherichia coli colonies. Observations indicate that bacterial growth preferentially occurs in the dextran phase, regardless of the microstructure. [ABSTRACT FROM AUTHOR]

Predictive microbiology has recently acknowledged the impact of the solid(like) food structure on microbial behavior. The presence of this solid(like) structure causes microorganisms to grow as colonies and no longer planktonically as in liquid. In this paper, the growth dynamics of Salmonella Typhimurium and Escherichia coli were studied as a function of temperature, considering different growth morphologies, i.e., (i) planktonic cells, (ii) immersed colonies and (iii) surface colonies. For all three growth morphologies, both microorganisms were grown in petri dishes. While E. coli was grown under optimal pH and water activity (aw), for S. Typhimurium pH and aw were adapted to 5.5 and 0.990. In order to mimic a solid(like) environment, 5% (w/v) gelatin was added. All petri dishes were incubated under static conditions at temperatures in the range [8.0°C-22.0°C]. Cell density was determined via viable plate counting. This work demonstrates that the growth morphology (planktonic vs. colony) has a negligible effect on the growth dynamics as a function of temperature. The observation of almost equal growth rates for planktonic cultures and colonies is in contrast to literature where, mostly, a difference is observed, i.e., μplanktonic cells≥μimmersed colonies≥μsurface colonies. This difference might be due to shaking of the liquid culture in these studies, which results in a nutrient and oxygen rich environment, in contrast to the diffusion-limited gel system. Experiments also indicate that lag phases for solid(like) systems are similar to those for the planktonic cultures, as can be found in literature for similar growth conditions. Considering the maximum cell density, no clear trend was deducted for either of the microorganisms. This study indicates that the growth parameters in the suboptimal temperature range do not depend on the growth morphology. For the considered experimental conditions, models previously developed for liquid environments can be used for solid(like) systems., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Sublethal injury (SI) poses major public health concerns since injured cells are responsible for serious limitations in food diagnostics and are susceptible to recovery, often developing adaptive stress responses. Detection of SI is based on the difference in plate counts between non-selective media, which represent the total cell population, and selective media, to which injured cells become sensitive. Selective media for detection of sublethal membrane damage are often based on NaCl supplement, although there is a lack of consensus in the literature about appropriate levels. Planktonic cells are generally used to investigate SI mechanisms, although they often exhibit different stress tolerance than cell colonies in/on solid food (model) systems. In this work, the effect of growth morphology, colony size and concentration of the gelling agent in the growth media, on the maximum non-inhibitory NaCl concentration in the plating medium was assessed for Escherichia coli, Salmonella Typhimurium and Listeria innocua. Stationary phase cultures of planktonic cells and large and small colonies grown in either 1.5% (w/v) xanthan gum-based system or 2.5% (w/v) xanthan gum-based system exhibited significantly different viable counts and osmotolerance. The effect of cell arrangement and xanthan gum percentage in the growth media depended on the microorganism under investigation. Additionally, differences in the maximum non-inhibitory concentration were evident, with 5.0% (w/v) NaCl for the Gram-negative bacteria and 6.5% (w/v), for L. innocua. Different extent of colony shrinkage and morphological damage was observed as NaCl concentration in the plating medium increased. This information will contribute to define NaCl-based selective media for accurate SI detection under realistic scenarios., (Copyright © 2014. Published by Elsevier Ltd.)

The occurrence of sublethally injured cells in foods poses major public health concerns and is an essential aspect when assessing the microbial response to food preservation strategies, yet there is limited research dealing with its specific implications for mild heating. All available studies so far have been performed in broths colonized by planktonic cells, although their susceptibility to lethal agents has often been reported to be markedly different to the stress tolerance of cell colonies developed in solid foods. In this work, the effect of planktonic and colony growth, as well as the influence of colony density on sublethal injury induced by mild heating of Escherichia coli, Salmonella Typhimurium and Listeria innocua were assessed in food model systems. Detection of injured survivors relied on their inability to form visible colonies on salt-based selective media, which do not affect the growth of healthy cells. Sublethal injury (SI) increased rapidly with shorter exposure times and afterwards, decreased progressively, suggesting a mechanism of cumulative damage triggering lethal instead of SI. Cell arrangement affected the degree of SI, higher values being generally found for gelified systems, although the effect of colony density depended on the target microorganism. This information is essential for optimizing the design of food safety assurance systems., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Up to now, it is generally observed that (i) the microbial growth domain is confined by structure-induced stress, or (ii) a solid(-like) environment can enhance microbial survival/growth. In most studies in solid(-like) systems, structure is induced by the addition of gelatin. The aim of this study was to evaluate the effect of other structure-inducing components on the growth dynamics. Both single and binary gel systems are used. Growth is studied when simultaneously exposed to salt stress. Experiments are performed in spectrophotometer tubes, filled with 1 mL of liquid, or structured inoculated brain heart infusion. Four different (combinations of) gelling agents are tested, that is, gelatin, xanthan gum, a 50% combination of xanthan gum and gelatin, and a 50% combination of carrageenan and gelatin. Experiments determine the growth behavior of both Escherichia coli (0% to 0.5% and 1%, 2%, 3%, 4%, and 5% NaCl) and Salmonella Typhimurium (0%, 1%, 2%, 3%, 4%, and 5% NaCl) at 23.5 and 27 °C. By means of plate counting, the growth dynamics are determined. At the studied conditions, growth of E. coli and Salmonella Typhimurium seems independent of the type of structure-inducing component. However, at higher concentrations of salt (>2%), lag phases are typically shorter in solid(-like) systems than in liquid media. For the conditions tested, the effect of a structured environment on growth rate and maximal cell density can be neglected., (© 2013 Institute of Food Technologists®)

Recently, the focus of predictive food microbiology has shifted towards more mechanistically-inspired modelling. Together with this trend, the need for methods that allow rapid data collection at the (intra)cellular level, as well as the intermediate subpopulation/colony level, has emerged. Although several experimental techniques are currently available to study colony dynamics in/on solid media, their widespread implementation as high-throughput methods remains a challenge. In this research, a novel method is presented to study colony growth based on optical density measurements performed in microtiter plates. An area scan procedure was applied to monitor individual Escherichia coli colonies in 48-well plates at 30 °C. Based on a fixed threshold value to separate the object (colony) from the background, the colony area was determined as a function of time. With this technique, expansion of the colony in radial direction could be monitored. Practical limitations (i.e., maximum achievable resolution and colony size) of the proposed method were investigated. A comparison was made with existing methods at the level of hardware requirements, data acquisition and data processing. Overall, the novel optical density method proved to be a flexible, high-throughput tool for monitoring (the mechanisms of) microbial colony growth in solid(like) systems., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The growth dynamics of Listeria monocytogenes strains isolated from salmon or a salmon processing environment and two reference Listeria innocua strains were investigated at refrigerated and close-to-optimal growth temperatures. Estimates for the growth rates and the lag-phase duration at 4, 8, 12, and 30°C were obtained for optical density measurements by using different growth parameter estimation methods, i.e., the serial dilution (SD) method and the relative rate to detection (RRD) method. Both single L. innocua and L. monocytogenes strains and mixtures of L. monocytogenes strains (cocktails) were studied. Both methods show an increase in maximum growth rate (μ(max)) of Listeria with increasing temperatures. Generally, single-strain growth rate estimates were quite similar for both species, although L. monocytogenes showed slightly higher μ(max) estimates at 4°C. The SD method gave the highest estimates for the growth rate, i.e., the estimates from the RRD method were 10 to 20% lower. This should lead to caution when using the latter method for Listeria, particularly at lower temperatures. Overall, the SD method is preferred as this method yields μ(max) estimates close to the biological value and provides estimates for the duration of lag time (λ). For discrimination between different strains, λ appeared to be a more suitable parameter than μ(max). This effect was most prominent for L. innocua. Significant differences were observed between μ(max) and/or λ of L. monocytogenes cocktails and single strains at all temperatures investigated. At 4°C, the average growth rate of cocktails was higher than that of single strains. At 8 and 30°C, this trend was reversed. The average λ of single strains were more than twice as long as those of cocktails at 4°C. At 8 and 30°C, the λ of cocktails were significantly slower than those of single strains, but the variation was considerably less and the differences were less pronounced.