Related Experiment Video
Updated: Aug 22, 2025
Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria
Published on: May 8, 2013
Characterization of Novel Klebsiella Phage PG14 and Its Antibiofilm Efficacy
Mansura S Mulani1,2, Shital N Kumkar1, Karishma R Pardesi1
1Department of Microbiology, Savitribai Phule Pune University, Pune, Maharashtra, India.
Abstract:
The increasing frequency of infections caused by multidrug-resistant Klebsiella pneumoniae demands the development of unconventional therapies. Here, we isolated, characterized, and sequenced a Klebsiella phage PG14 that infects and lyses carbapenem-resistant K. pneumoniae G14. Phage PG14 showed morphology similar to the phages belonging to the family Siphoviridae. The adsorption curve of phage PG14 showed more than 90% adsorption of phages on a host within 12 min. A latent period of 20 min and a burst size of 47 was observed in the one step growth curve. Phage PG14 is stable at a temperature below 30°C and in the pH range of 6 to 8. The PG14 genome showed no putative genes associated with virulence and antibiotic resistance. Additionally, it has shown lysis against 6 out of 13 isolates tested, suggesting the suitability of this phage for therapeutic applications. Phage PG14 showed more than a 7-log cycle reduction in K. pneumoniae planktonic cells after 24 h of treatment at a multiplicity of infection (MOI) of 10. The phage PG14 showed a significant inhibition and disruption of biofilm produced by K. pneumoniae G14. The promising results of this study nominate phage PG14 as a potential candidate for phage therapy. IMPORTANCE Klebsiella pneumoniae is one of the members of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) group of pathogens and is responsible for nosocomial infections. The global increase of carbapenem-resistant K. pneumoniae has developed a substantial clinical threat because of the dearth of therapeutic choices available. K. pneumoniae is one of the commonly found bacteria responsible for biofilm-related infections. Due to the inherent tolerance of biofilms to antibiotics, there is a growing need to develop alternative strategies to control biofilm-associated infections. This study characterized a novel bacteriophage PG14, which can inhibit and disrupt the K. pneumoniae biofilm. The genome of phage PG14 does not show any putative genes related to antimicrobial resistance or virulence, making it a potential candidate for phage therapy. This study displays the possibility of treating infections caused by multidrug-resistant (MDR) isolates of K. pneumoniae using phage PG14 alone or combined with other therapeutic agents.
Insights
A novel bacteriophage, PG14, effectively lyses multidrug-resistant Klebsiella pneumoniae, including carbapenem-resistant strains. This phage shows potential for treating infections and disrupting biofilms, offering a promising alternative to antibiotics.
Area of Science:
- Microbiology
- Virology
- Biotechnology
Background:
- Multidrug-resistant Klebsiella pneumoniae poses a significant threat due to increasing infections and limited treatment options.
- Klebsiella pneumoniae is a key member of the ESKAPE group of pathogens, frequently causing nosocomial infections.
- Biofilm-associated infections caused by K. pneumoniae are challenging to treat with conventional antibiotics.
Purpose of the Study:
- To isolate, characterize, and sequence a bacteriophage targeting multidrug-resistant Klebsiella pneumoniae.
- To evaluate the efficacy of the isolated phage PG14 against planktonic and biofilm forms of K. pneumoniae.
- To assess the safety profile of phage PG14 for potential therapeutic applications.
Main Methods:
- Isolation and characterization of Klebsiella phage PG14.
- Morphological analysis and genomic sequencing of phage PG14.
- One-step growth curve analysis to determine latent period and burst size.
- Adsorption kinetics, stability testing (temperature and pH), and lytic activity assessment against K. pneumoniae isolates.
- Evaluation of phage PG14's efficacy against planktonic cells and biofilms.
Main Results:
- Phage PG14, resembling the Siphoviridae family, efficiently adsorbed to its host within 12 minutes.
- The phage exhibited a latent period of 20 minutes and a burst size of 47, with optimal stability below 30°C and pH 6-8.
- Phage PG14 demonstrated significant reduction (over 7-log cycles) of planktonic K. pneumoniae and effectively inhibited/disrupted biofilms.
- Genomic analysis revealed no genes associated with virulence or antibiotic resistance in phage PG14.
- The phage showed lytic activity against 6 out of 13 tested K. pneumoniae isolates.
Conclusions:
- Phage PG14 is a promising candidate for phage therapy against multidrug-resistant Klebsiella pneumoniae infections.
- The phage's ability to inhibit and disrupt biofilms makes it valuable for treating persistent infections.
- The absence of virulence and antibiotic resistance genes in its genome supports the safety of phage PG14 for therapeutic use.
