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Published on: December 23, 2022
Characterization and complete genome sequence analysis of novel bacteriophage IME-EFm1 infecting Enterococcus faecium
Yahui Wang1,2, Wei Wang3,1, Yongqiang Lv4
1State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, PR China.
Abstract:
We isolated and characterized a novel virulent bacteriophage, IME-EFm1, specifically infecting multidrug-resistant Enterococcus faecium. IME-EFm1 is morphologically similar to members of the family Siphoviridae. It was found capable of lysing a wide range of our E. faecium collections, including two strains resistant to vancomycin. One-step growth tests revealed the host lysis activity of phage IME-EFm1, with a latent time of 30 min and a large burst size of 116 p.f.u. per cell. These biological characteristics suggested that IME-EFm1 has the potential to be used as a therapeutic agent. The complete genome of IME-EFm1 was 42 597 bp, and was linear, with terminally non-redundant dsDNA and a G+C content of 35.2 mol%. The termini of the phage genome were determined with next-generation sequencing and were further confirmed by nuclease digestion analysis. To our knowledge, this is the first report of a complete genome sequence of a bacteriophage infecting E. faecium. IME-EFm1 exhibited a low similarity to other phages in terms of genome organization and structural protein amino acid sequences. The coding region corresponded to 90.7 % of the genome; 70 putative ORFs were deduced and, of these, 29 could be functionally identified based on their homology to previously characterized proteins. A predicted metallo-β-lactamase gene was detected in the genome sequence. The identification of an antibiotic resistance gene emphasizes the necessity for complete genome sequencing of a phage to ensure it is free of any undesirable genes before use as a therapeutic agent against bacterial pathogens.
Insights
A novel bacteriophage, IME-EFm1, effectively lyses multidrug-resistant Enterococcus faecium, including vancomycin-resistant strains. Its complete genome sequencing reveals potential therapeutic applications, highlighting the importance of phage screening for antibiotic resistance genes.
Area of Science:
- Microbiology
- Genomics
- Phage Therapy
Background:
- Multidrug-resistant Enterococcus faecium poses a significant clinical challenge.
- Bacteriophages offer a promising alternative to antibiotics for combating resistant bacteria.
Purpose of the Study:
- To isolate and characterize a novel bacteriophage targeting multidrug-resistant Enterococcus faecium.
- To determine the complete genome sequence of the isolated bacteriophage and assess its therapeutic potential.
Main Methods:
- Isolation and morphological characterization of bacteriophage IME-EFm1.
- One-step growth analysis to determine lytic activity.
- Whole-genome sequencing and bioinformatic analysis.
Main Results:
- Bacteriophage IME-EFm1 effectively lysed various Enterococcus faecium strains, including vancomycin-resistant ones.
- The phage exhibits a latent period of 30 min and a burst size of 116 p.f.u./cell.
- The complete 42,597 bp genome sequence revealed 70 putative open reading frames, with one encoding a metallo-β-lactamase gene.
Conclusions:
- Bacteriophage IME-EFm1 is a virulent phage with potential for therapeutic use against multidrug-resistant Enterococcus faecium.
- Complete genome sequencing is crucial to identify and exclude undesirable genes, such as antibiotic resistance genes, before therapeutic application.
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