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Bottom-up assembly of beneficial multi-species biofilms targeting undesirable bacteria using 3D fluorescence imaging. Bottom-up assembly of beneficial multi-species biofilms targeting undesirable bacteria using 3D fluorescence imaging

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Guéneau, Virgile | Guillier, Laurent | Berdous, Cécile | Rubrice, Raphael | Carlioz, Antoine | Cornuéjols, Antoine | Guigue, Vincent | Serror, Pascale | Noirot-Gros, Marie-Françoise | Castex, Mathieu | Briandet, Romain

Edité par CCSD -

International audience. Engineered synthetic microbial communities (SynComs) forming beneficial biofilms offer a promising strategy in biotechnology to prevent harmful bacterial settlement and reduce reliance on chemical antimicrobials. However, strain selection criteria and antagonistic mechanisms remain unclear. This study presents a bottom-up approach integrating 3D fluorescence imaging with high-throughput analysis of multi-strain biofilms. Our findings reveal that competitive strains against undesirable bacteria may also exclude beneficial ones, highlighting the need for compatibility control in SynCom assembly. SynComs composed of B. velezensis and Pediococcus spp. enhanced pathogen exclusion compared to single strains. Temporal analysis of biofilm interactions, supported by mathematical models, showed that pathogen exclusion was primarily driven by nutritional competition (Jameson effect) with additional specific interference mechanisms (prey-predator Lotka-Volterra model). Furthermore, pre-established SynComs significantly increased pathogen inhibition, indicating a distinct biofilm-associated exclusion effect. These insights provide a framework for SynCom assembly and refine our understanding of mechanisms driving beneficial biofilm applications.

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