Estefania Noriega Fernandez

Introducing a novel interaction model structure for the combined effect of temperature and pH on the microbial growth rate.

Efficient modelling of the microbial growth rate can be performed by combining the effects of individual conditions in a multiplicative way, known as the gamma concept. However, several studies have illustrated that interactions between different effects should be taken into account at stressing environmental conditions to achieve a more accurate description of the growth rate. In this research, a novel approach for modeling the interactions between the effects of environmental conditions on the microbial growth rate is introduced. As a case study, the effect of temperature and pH on the growth rate of Escherichia coli K12 is modeled, based on a set of computer controlled bioreactor experiments performed under static environmental conditions. The models compared in this case study are the gamma model, the model of Augustin and Carlier (2000), the model of Le Marc et al. (2002) and the novel multiplicative interaction model, developed in this paper. This novel model enables the separate identification of interactions between the effects of two (or more) environmental conditions. The comparison of these models focuses on the accuracy, interpretability and compatibility with efficient modeling approaches. Moreover, for the separate effects of temperature and pH, new cardinal parameter model structures are proposed. The novel interaction model contributes to a generic modeling approach, resulting in predictive models that are (i) accurate, (ii) easily identifiable with a limited work load, (iii) modular, and (iv) biologically interpretable.

Recent trends in non-invasive in situ techniques to monitor bacterial colonies in solid (model) food

Planktonic cells typically found in liquid systems, are routinely used for building predictive models or assessing the efficacy of food preserving technologies. However, freely suspended cells often show different susceptibility to environmental hurdles than colony cells in solid matrices. Limited oxygen, water and nutrient availability, metabolite accumulation and physical constraints due to cell immobilization in the matrix, are main factors affecting cell growth. Moreover, intra- and inter-colony interactions, as a consequence of the initial microbial load in solid systems, may affect microbial physiology. Predictive food microbiology approaches are moving toward a more realistic resemblance to food products, performing studies in structured solid systems instead of liquids. Since structured systems promote microbial cells to become immobilized and grow as colonies, it is essential to study the colony behavior, not only for food safety assurance systems, but also for understanding cell physiology and optimizing food production processes in solid matrices. Traditionally, microbial dynamics in solid systems have been assessed with a macroscopic approach by applying invasive analytical techniques; for instance, viable plate counting, which yield information about overall population. In the last years, this approach is being substituted by more mechanistically inspired ones at mesoscopic (colony) and microscopic (cell) levels. Therefore, non-invasive and in situ monitoring is mandatory for a deeper insight into bacterial colony dynamics. Several methodologies that enable high-throughput data collection have been developed, such as microscopy-based techniques coupled with image analysis and OD-based measurements in microplate readers. This research paper provides an overview of non-invasive in situ techniques to monitor bacterial colonies in solid (model) food and emphasizes their advantages and inconveniences in terms of accuracy, performance and output information.

An individual-based modeling approach to simulate the effects of cellular nutrient competition on Escherichia coli K-12 MG1655 colony behavior and interactions in aerobic structured food systems

Traditional kinetic models in predictive microbiology reliably predict macroscopic dynamics of planktonically-growing cell cultures in homogeneous liquid food systems. However, most food products have a semi-solid structure, where microorganisms grow locally in colonies. Individual colony cells exhibit strongly different and non-normally distributed behavior due to local nutrient competition. As a result, traditional models considering average population behavior in a homogeneous system do not describe colony dynamics in full detail. To incorporate local resource competition and individual cell differences, an individual-based modeling approach has been applied to Escherichia coli K-12 MG1655 colonies, considering the microbial cell as modeling unit. The first contribution of this individual-based model is to describe single colony growth under nutrient-deprived conditions. More specifically, the linear and stationary phase in the evolution of the colony radius, the evolution from a disk-like to branching morphology, and the emergence of a starvation zone in the colony center are simulated and compared to available experimental data. These phenomena occur earlier at more severe nutrient depletion conditions, i.e., at lower nutrient diffusivity and initial nutrient concentration in the medium. Furthermore, intercolony interactions have been simulated. Higher inoculum densities lead to stronger intercolony interactions, such as colony merging and smaller colony sizes, due to nutrient competition. This individual-based model contributes to the elucidation of characteristic experimentally observed colony behavior from mechanistic information about cellular physiology and interactions.