Environmental redox conditions and strain variation define phenazine-mediated antagonism in co-infecting bacteria

by Katlyn Todd, Olivia Schneider, Joshua M. Lawrence, Josefina L. Aronoff, Bartosz Witek, Valerie Velázquez-Colón, Verónica Santana-Ufret, Nicole L. Anderson, Krista Gunter, Moraima Noda, Ryan F. Relich, Lifan Zeng, Dominique H. Limoli, Christopher Whidbey, Jay Vornhagen

Pseudomonas aeruginosa and Klebsiella pneumoniae are gram-negative opportunistic pathogens that frequently colonize the human body and are major causes of infection. These bacteria are often co-isolated in polymicrobial urinary tract and lung infections, the latter of which is associated with increased disease severity and worse clinical outcomes. Despite their overlapping niches and clinical relevance, little is known about how these two pathogens interact and how those interactions influence human health. Given the growing recognition that microbial interactions are key drivers of disease, we investigated how P. aeruginosa and K. pneumoniae influence one another. We discovered an antagonistic interaction in which P. aeruginosa restricts the growth of K. pneumoniae. This inhibition is driven by phenazine production in P. aeruginosa, specifically the secondary metabolites pyocyanin and pyorubin, which are both necessary and sufficient to suppress K. pneumoniae growth. Using a diverse set of clinical isolates, we found that this antagonism is strain-dependent. Both the susceptibility of K. pneumoniae to phenazines and the ability of P. aeruginosa to restrict K. pneumoniae growth varies between strains. Moreover, the necessity of phenazine production is specific to the site of infection. Together, these findings demonstrate that strain background and environmental context are critical determinants of pathogen interactions. These findings reveal that both strain background and environmental redox conditions govern the ecological rules of pathogen interaction, providing a framework for predicting outcomes.