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Spatial structure over scales ranging from nanometres to centimetres (and beyond) varies markedly in diverse habitats and the industry-relevant settings that support microbial activity. Developing an understanding of the interplay between a structured environment and the associated microbial processes and ecology is fundamental, but challenging. Several novel approaches have recently been developed and implemented to help address key questions for the field: from the use of imaging tools such as X-ray Computed Tomography to explore microbial growth in soils, to the fabrication of scratched materials to examine microbial-surface interactions, to the design of microfluidic devices to track microbial biofilm formation and the metabolic processes therein. This review discusses new approaches and challenges for incorporating structured elements into the study of microbial processes across different scales. We highlight how such methods can be pivotal for furthering our understanding of microbial interactions with their environments.

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The physical environments in which microorganisms naturally reside rarely have homogeneous structure, and changes in their porous architecture can have a profound effect on microbial activities – effects that are not typically captured in conventional laboratory studies. Here, to investigate the influence of environmental structure on microbial responses to stress, we constructed structured environments with different pore properties (determined by X-ray Computed Tomography). First, using glass beads in different arrangements and inoculated with the soil yeast Saitozyma podzolica , increases in the average equivalent spherical diameters (ESD) of a structure’s porous architecture led to decreased survival of the yeast under a toxic metal challenge. This relationship was reproduced when yeasts were introduced into additively-manufactured lattice structures, comprising regular arrays with ESDs comparable to those of the bead structures. The pore ESD-dependency of metal resistance was not attributable to differences in cell density in micro-environments delimited by different pore sizes, supporting the inference that pore size specifically is the important parameter here in determining microbial survival of stress. These findings highlight the importance of the physical architecture of an organism’s immediate environment for its response to environmental perturbation, while offering new tools for investigating these interactions in the laboratory. IMPORTANCE Interactions between cells and their structured environments are poorly understood but have significant implications for organismal success in both natural and non-natural settings. This work uses a multidisciplinary approach to develop laboratory models with which the influence of a key parameter of environmental structure – pore size – on cell activities can be dissected. Using these new methods in tandem with additive manufacturing, we demonstrate that resistance of yeast soil-isolates to stress (from a common metal pollutant) is inversely related to pore size of their environment. This has important ramifications for understanding how microorganisms respond to stress in different environments. The findings also establish new pathways for resolving the effects of physical environment on microbial activity, enabling important understanding that is not readily attainable with traditional bulk-sampling and analysis approaches.

The weak acid sorbic acid is a common preservative used in soft drink beverages to control microbial spoilage. Consumers and industry are increasingly transitioning to low-sugar food formulations, but potential impacts of reduced-sugar on preservative efficacy are barely characterised. In this study, we report enhanced sorbic acid resistance of spoilage yeasts in low-glucose conditions. We had anticipated that low glucose may induce respiratory metabolism, previously shown to be targeted by sorbic acid. However, a shift from respiratory to fermentative metabolism was correlated with the sorbic acid resistance in low glucose. Fermentation-deficient yeast species did not show the low-glucose resistance phenotype. Phenotypes observed for certain yeast deletion strains suggested roles for glucose signalling and repression pathways in the sorbic acid resistance at low glucose. This low-glucose induced sorbic acid resistance was alleviated by supplementing yeast cultures with succinic acid, a metabolic intermediate of respiratory metabolism (and a food-safe additive) that promoted respiration. The results indicate that metabolic adaptation of spoilage yeasts promotes sorbic acid resistance at low glucose, providing new insight into potential spoilage, and preservation, of foodstuffs as both food producers and consumers move towards a reduced-sugar landscape.