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Anti-virulent Disruption of Pathogenic Biofilms using Engineered Quorum-quenching Lactonases
Published on: January 1, 2016
A K-17 serotype specific Klebsiella phage JKP2 with biofilm reduction potential
Muhammad Asif1, Iqbal Ahmad Alvi2, Muhammad Waqas3
1Institute of Microbiology and Molecular Genetics, University of the Punjab, Lahore, Pakistan; Department of Pathology, King Edward Medical University, Lahore, Pakistan.
Abstract:
Klebsiella pneumoniae is an opportunistic pathogen responsible for nearly one-third of all Gram-negative infections. Increasing antibiotic resistance has pushed scientists to look for alternative therapeutics. Bacteriophages have emerged as one of the promising alternatives. In the current study, the Klebsiella phage JKP2 was isolated from a sewage sample and characterized against the K-17 serotype of K. pneumoniae. It produced bulls-eye-shaped clear plaques and has a latent period of 45 min with a burst size of 70 pfu/cell. It remained stable at tested pH (5 to 10) and temperatures (37 to 60 °C). Its optimum temperature for long-term storage is 4 °C and -80 °C. The JKP2 showed its infectivity against the K. pneumoniae K-17 serotype only. It controlled planktonic cells of K. pneumoniae 12 h post-incubation. At MOI-1, it efficiently eliminated 98% of 24 and 96% of 48-hour-old biofilm and 86% and 82% of mature biofilm of day 3 and 4, respectively. The JKP2 has an icosahedral capsid of 54 ± 0.5 nm with a short, non-contractile tail, measuring 12 ± 0.2 nm. It possesses a double-stranded DNA genome of 43.2 kbp with 54.1% GC content and encodes 54 proteins, including 29 with known functions and 25 with unknown functions. JKP2 was classified as Drulisvirus within the Autographiviridae family. It uses a T7-like direct terminal repeat strategy for genome packaging. JKP2 can be applied safely for therapeutic purposes as it does not encode an integrase or repressor genes, antibiotic resistance genes, bacterial virulence factors, and mycotoxins.
Insights
Klebsiella phage JKP2 effectively targets Klebsiella pneumoniae, an opportunistic pathogen. This bacteriophage shows stability and safety for therapeutic use against antibiotic-resistant Gram-negative infections.
Area of Science:
- Microbiology
- Virology
- Infectious Diseases
Background:
- Klebsiella pneumoniae is a major cause of Gram-negative infections, with rising antibiotic resistance necessitating alternative treatments.
- Bacteriophages (phages) are viruses that infect bacteria and represent a promising alternative therapeutic strategy.
Purpose of the Study:
- To isolate and characterize Klebsiella phage JKP2 for its potential as a therapeutic agent against Klebsiella pneumoniae.
Main Methods:
- Isolation of phage JKP2 from sewage.
- Characterization of phage morphology, stability (pH, temperature), and infectivity against K. pneumoniae K-17 serotype.
- Assessment of efficacy against planktonic cells and biofilms.
- Genomic analysis and phylogenetic classification.
Main Results:
- Phage JKP2 demonstrated stability across a pH range of 5-10 and temperatures of 37-60°C, with optimal storage at 4°C and -80°C.
- JKP2 effectively reduced planktonic K. pneumoniae within 12 hours and significantly degraded 24- to 4-day-old biofilms.
- Genomic analysis revealed JKP2 belongs to the Drulisvirus genus (Autographiviridae family), possesses a dsDNA genome, and lacks genes for integrase, repressor, antibiotic resistance, virulence factors, or mycotoxins.
Conclusions:
- Klebsiella phage JKP2 is a robust and specific bacteriophage with significant potential for therapeutic applications against Klebsiella pneumoniae infections.
- Its safety profile, demonstrated by the absence of undesirable genes, supports its consideration for clinical use in combating antibiotic-resistant bacteria.
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