Heat Stress Induced Bacterial Tolerance against Phage Facilitates the Evolution of Resistance

Abstract Antibiotic resistance and tolerance present significant challenges in global healthcare, necessitating alternative strategies such as phage therapy. However, the rapid emergence of phage-resistant mutants poses a potential risk. Here, we investigated bacterial persistence against phages, characterized by heterogeneous survival, analogous to antibiotic persistence. We found that heat treatment enhanced persistence and increase bacterial survival under phage exposure, subsequently promoting the evolution of phage resistance. Further experiments demonstrated that heat stress leads to a reduction in envelope components, thereby inhibiting phage DNA injection. Additionally, this heat-induced reduction resulted in systematic alterations in envelope stress responses, rendering bacteria tolerant to the antibiotic polymyxin while making them hypersensitive to pH changes and immune clearance. Our findings provide novel insights into bacteria-phage interactions and highlight potential challenges in implementing phage therapy in clinical settings.