Polymicrobial catheter biofilms sustain susceptible Enterococcus faecalis and Escherichia coli during β-lactam treatment

Broad-spectrum β-lactam exposure can select for Enterococcus -dominated urinary communities in catheterized intensive-care patients, even when co-colonizing Escherichia coli remains susceptible. We investigated paired E. faecalis and E. coli isolates recovered before and after piperacillin-tazobactam (TZP) treatment using a catheter biofilm model and showed that their survival depends on mutualism and biofilm-dependent persistence. Without antibiotics, E. faecalis reduced E. coli biofilm formation yet promoted pre-attachment coaggregation and reorganized mixed-biofilm architecture on the catheter. Despite TZP susceptibility and the absence of resistance determinants, catheter-associated biofilms and biofilm-dispersed cells survived concentrations 250- to 1000-fold above their MICs, whereas planktonic cells were eliminated. Survivors retained susceptibility but showed delayed regrowth, consistent with a transient persister-like state. In the post-treatment pair, each species sustained the other during recovery, coinciding with a nonsynonymous substitution in the enterococcal surface adhesin Esp. These findings show that antagonistic and cooperative interactions can coexist within catheter biofilms and enable susceptible polymicrobial communities to withstand β-lactam treatment without β-lactam resistance.