Updated phage banks essential to cope with pathogen evolution: Lessons from Klebsiella pneumoniae and their phages.

Background: Multidrug-resistant Klebsiella pneumoniae (MDR-KP) leads global health concerns as an infectious agent due to many virulence factors, including biofilm formation. The growing urgency for alternative treatment strategies beyond antibiotics has renewed interest in bacteriophages. Pathogen evolution is dynamic and can lead to phage resistance.

Methods: Bacteriophages were isolated from environmental sources and screened against a panel of 280 MDR- K. pneumoniae clinical isolates (74 from 2018 to 2020 and 167 from 2022-23 and 39 environmental KP isolates). Phages were grouped by host range and DNA fingerprinting. Five candidate phages with unique and broad host coverage were selected for further characterization and genome sequencing.

Results: Five candidate phages exhibited diverse host range patterns, strong bacteriolytic activity and significant antibiofilm activity even at low multiplicity of infection. Whole genome sequencing analysis revealed phage KPØ6 to be a novel Taipevirus species with low intergenomic similarity to known phages, and it showed the broadest host range on isolates from the 2018-2020 panel. A significant rise in phage resistance among MDR-KP isolate panels of 2018-2020 and 2022-2023 was observed.

Conclusion: Lytic bacteriophages offer a promising alternative for tackling MDR-KP infections, especially within healthcare environments. The phages characterized in this study demonstrate strong potential for both biocontrol and therapeutic use against MDR-KP, including infections involving biofilms. However, the dynamic nature of bacterial evolution over five years reiterates the need to update phage banks.