Influence of interspecies interactions on the spatial organization of dual species bacterial communities

Interspecies interactions in bacterial biofilms have important impacts on the composition and function of communities in natural and engineered systems. To investigate these interactions, synthetic communities provide experimentally tractable systems. Biofilms grown on agar-surfaces have been used for investigating the eco-evolutionary and biophysical forces that determine community composition and spatial distribution of bacteria. Prior studies have used genetically identical bacterial strains and strains with specific mutations, that express different fluorescent proteins, to investigate intraspecies interactions. Here, we investigated interspecies interactions and, specifically, determined the community composition and spatial distribution in synthetic communities of Pseudomonas aeruginosa, Pseudomonas protegens and Klebsiella pneumoniae. Using quantitative microscopic imaging, we found that interspecies interactions in multispecies colonies were influenced by type IV pilus mediated motility, extracellular matrix secretion, environmental parameters, and these effects were also influenced by the specific partner in the dual species combinations. These results indicate that the patterns observable in mixed species colonies can be used to understand the mechanisms that drive interspecies interactions, which are dependent on the interplay between specific species’ physiology and environmental conditions. •Spatial patterns in bacterial colonies are species and interaction dependent.•Surface motility and extracellar matrix production affect interspecies interactions.•Agar surface colonies show how bacteria interact in biofilms.