Organ agar serves as physiologically relevant alternative for in vivo bacterial colonization

Animal models for host-microbial interactions have proven valuable, yielding physiologically relevant data that may be otherwise difficult to obtain. Unfortunately, such models are lacking or nonexistent for many microbes. Here, we introduce organ agar, a straightforward method to enable the screening of large mutant libraries while avoiding physiological bottlenecks. We demonstrate that growth defects on organ agar were translatable to bacterial colonization deficiencies in a murine model. Specifically, we present a urinary tract infection agar model to interrogate an ordered library of Proteus mirabilis transposon mutants, with accurate prediction of bacterial genes critical for host colonization. Thus, we demonstrate the ability of ex vivo organ agar to reproduce in vivo deficiencies. Organ agar was also useful for identifying previously unknown links between biosynthetic genes and swarming motility. This work provides a readily adoptable technique that is economical and uses substantially fewer animals. We anticipate this method will be useful for a wide variety of microorganisms, both pathogenic and commensal, in a diverse range of model host species.

Automated summary

Animal models for host-microbial interactions are valuable but lacking for many microbes. This study introduces a method called organ agar, which enables screening of mutant libraries without physiological bottlenecks. The authors demonstrate that growth defects on organ agar accurately predict bacterial colonization deficiencies in a murine model. The method is also useful for identifying unknown links between biosynthetic genes and swarming motility.