Keystone engineering enables collective range expansion in microbial communities

Abstract Keystone engineers profoundly influence microbial communities by altering their shared environment, often by modifying key resources. Here, we show that in an antibiotic-treated microbial community, bacterial spread is controlled by keystone engineering affecting dispersal—an effect hidden in well-mixed environments. Focusing on two pathogens, non-motile Klebsiella pneumoniae and motile Pseudomonas aeruginosa, we found that both tolerate a β-lactam antibiotic, with Pseudomonas being more resilient and dominating in well-mixed cultures. During range expansion, however, the antibiotic inhibits Pseudomonas’ ability to spread unless it is near Klebsiella—Klebsiella degrades the antibiotic to create a “clear zone” that allows Pseudomonas to expand, at the expense of Klebsiella’s own growth, thus acting as a keystone engineer. As Pseudomonas spreads, it competitively suppresses Klebsiella. Our modeling and experimental analyses reveal that this keystone effect operates at a millimeter scale. We also observed similar keystone engineering by a Bacillus species isolated from a hospital sink, in both pairwise and eight-member bacterial communities with its co-isolates. These findings suggest that spatially explicit experiments are essential to understand certain keystone engineering mechanisms and have implications for surface-associated microbial communities like biofilms, as well as for diagnosing and treating polymicrobial infections involving drug-degrading, non-motile (e.g., Klebsiella), and drug-tolerant, motile (e.g., Pseudomonas) bacteria. Competing Interest Statement The authors have declared no competing interest. Footnotes Text updated to clarify terminology. Main figures updated for enhanced clarity. New Supplementary Figures and Tables were added to address reviewers' comments. Corresponding authors information updated.