Spatio-temporal modeling of the crowding conditions and metabolic variability in microbial communities

The metabolic capabilities of the species and the local environment shape the microbial interactions in a community either through the exchange of metabolic products or the competition for the resources. Cells are often arranged in close proximity to each other, creating a crowded environment that unevenly reduce the diffusion of nutrients. Herein, we investigated how the crowding conditions and metabolic variability among cells shape the dynamics of microbial communities. For this, we developed CROMICS, a spatio-temporal framework that combines techniques such as individual-based modeling, scaled particle theory, and thermodynamic flux analysis to explicitly incorporate the cell metabolism and the impact of the presence of macromolecular components on the nutrients diffusion. This framework was used to study two archetypical microbial communities (i) Escherichia coli and Salmonella enterica that cooperate with each other by exchanging metabolites, and (ii) two E. coli with different production level of extracellular polymeric substances (EPS) that compete for the same nutrients. In the mutualistic community, our results demonstrate that crowding enhanced the fitness of cooperative mutants by reducing the leakage of metabolites from the region where they are produced, avoiding the resource competition with non-cooperative cells. Moreover, we also show that E. coli EPS-secreting mutants won the competition against the non-secreting cells by creating less dense structures (i.e. increasing the spacing among the cells) that allow mutants to expand and reach regions closer to the nutrient supply point. A modest enhancement of the relative fitness of EPS-secreting cells over the non-secreting ones were found when the crowding effect was taken into account in the simulations. The emergence of cell-cell interactions and the intracellular conflicts arising from the trade-off between growth and the secretion of metabolites or EPS could provide a local competitive advantage to one species, either by supplying more cross-feeding metabolites or by creating a less dense neighborhood. Author summary Microbial communities play a key role in biogeochemical cycles, bioremediation, and human health. In crowded microbial systems such as biofilms and cellular aggregates, the close proximity between individual cells reduces the free space for the nutrients diffusion. To model the heterogeneous nature of these microbial systems, we developed CROMICS, a framework that integrates the information about the metabolic capabilities of each individual cell as well as the size and location of cells and macromolecules in the medium. The interactions among the individuals arise naturally through competition for or the exchange of metabolites. We show how the presence of mutants and a reduced diffusion in crowded environments can perturb the local availability of nutrients and therefore modify the dynamics of a microbial community. The discovered mechanisms underlying the microbial interactions in crowded systems together with the developed framework represent a valuable starting point for future studies of the interplay of human microbiome and host metabolism, the pathogen invasion, and the evaluation of antibiotic effectiveness.