Isolation and characterization of bacteriophage targeting multidrug-resistant Klebsiella pneumoniae from wastewater

Abstract

The emergence of multidrug-resistant Klebsiella pneumoniae has become a major global concern due to its rapid dissemination and limited treatment options. This study aimed to isolate and characterize bacteriophages from wastewater samples targeting MDR K. pneumoniae. Blue colonies on HiCrome KPC agar indicated carbapenemase-producing isolates. Six distinct lytic phages were recovered using the double-layer agar method, and among them, phage 1.3B exhibited the most stable and consistent lytic profile. The phage was tested for stability across various NaCl concentrations (0.5–5%), demonstrating optimal activity at 0.9% salinity. PCR confirmed the presence of resistance-associated genes, including blaNDM, and blaTEM and virulent gene iroB. These findings highlight the potential of naturally occurring phages as alternative biocontrol agents against MDR K. pneumoniae and provide a foundation for further genomic and therapeutic evaluation.