Potential of a phage cocktail in the treatment of multidrug-resistant Klebsiella pneumoniae pulmonary infection in mice

The emergence of multidrug-resistant Klebsiella pneumoniae, including carbapenem-resistant K. pneumoniae (CRKP), as one of the most common and notable superbugs, has long been a major threat to public health. As natural predators of bacteria, bacteriophages (or phages) can induce the lysis of bacterial cells. Herein, we report the isolation and characterization of two phages and their efficacy in the control of CRKP. Using the sequence type 11 (ST11) CRKP strain THR60 and its related strain THR60r as the host bacteria, phages GZ7 and GZ9 were isolated from hospital sewage, respectively. GZ7 is a myovirus with a head of 64 nm in diameter and a tail of 97 nm in length, and GZ9 is a siphovirus with a head of 67 nm in diameter and a tail of 175 nm in length. The host spectrum of a phage cocktail consisting of phages GZ7 and GZ9 was 82.4% (42/51 strains). An in vitro antibacterial activity assay demonstrated that the phage cocktail consisting of GZ7 and GZ9 effectively inhibited bacterial growth and suppressed the production of phage-resistant bacteria. In vivo experiment revealed that phage-treated mice exhibited lower K. pneumoniae burdens in the lungs compared to untreated control mice. Additionally, phage-treated mice experienced less body weight loss and had reduced levels of inflammatory cytokines in their lungs. Lung lesion conditions were significantly improved by phage therapy. Notably, the therapeutic effects of the GZ7 + GZ9 cocktail and GZ7 alone on mouse pulmonary infections were nearly equivalent. Therefore, phages GZ7 and GZ9 showed potential as alternatives to antibiotics for treating pneumonia caused by multidrug-resistant K. pneumoniae.