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Published on: October 14, 2011
Characterization and genome analysis of novel phage vB_EfaP_IME195 infecting Enterococcus faecalis
Ronghuan Wang1, Shaozhen Xing2, Feiyang Zhao3
1School of Public Health, Lanzhou University, Lanzhou, 730000, China.
Abstract:
Enterococcus faecalis is one of the main bacteria in the human and animal intestine but is also classed as an opportunistic pathogen. During normal growth, E. faecalis produces natural antibiotics and is conducive to human health. As ectopic parasites, E. faecalis is capable of causing infective endocarditis, neonatal sepsis, bloodstream infections, bacteremia, and intraabdominal infections. With the incidence of antibiotic resistance reaching crisis point, it is imperative to find alternative treatments for multidrug-resistant infections. Using phage for pathogen control is a promising treatment option to combat bacterial resistance. In this study, a lytic phage, designated vB_EfaP_IME195, was isolated from hospital sewage using a clinical multidrug-resistant Enterococcus faecalis strain as an indicator. The one-step growth curve with the optimal multiplicity of infection of (MOI) 0.01 revealed a latent period of ~ 30 min and a burst size of ~ 120 plaque-forming units (pfu) per cell. Transmission electron microscopy showed that the phage belongs to the family Podoviridae. Phage vB_EfaP_IME195 has a linear, double-stranded DNA genome of 18,607 bp with a G + C content of 33% and 27 coding sequences (GenBank accession no. KT932700). Run-off sequencing experiments showed that the phage has a unique 59-bp inverted repeat sequences at the terminal ends. BLASTn analysis revealed that vB_EfaP_IME195 shares 92% identity (93% genome coverage) with unpublished E. faecalis phage Idefix. This study reported a novel E. faecalis phage with unique genome termini containing inverted repeats. The isolation and characterization of this novel lytic E. faecalis phage provides the basis for the development of new therapeutic agents like phage cocktails for multidrug-resistant E. faecalis infection, and its unique genomic feature would also provide valuable knowledge and insight for further phage genome analysis.
Insights
Researchers isolated a novel lytic phage, vB_EfaP_IME195, effective against multidrug-resistant Enterococcus faecalis. This phage, belonging to the Podoviridae family, shows potential for developing new phage therapies against resistant bacterial infections.
Area of Science:
- Microbiology
- Bacteriology
- Virology
Background:
- Enterococcus faecalis is a common gut bacterium and an opportunistic pathogen causing severe infections.
- Rising antibiotic resistance necessitates alternative treatments for multidrug-resistant Enterococcus faecalis infections.
- Bacteriophage therapy presents a promising strategy to combat antibiotic-resistant bacteria.
Purpose of the Study:
- To isolate and characterize a novel lytic bacteriophage targeting multidrug-resistant Enterococcus faecalis.
- To evaluate the phage's biological and genomic properties for potential therapeutic applications.
Main Methods:
- Isolation of lytic phage vB_EfaP_IME195 from hospital sewage using a multidrug-resistant Enterococcus faecalis strain.
- One-step growth curve analysis to determine latent period and burst size.
- Transmission electron microscopy for morphological classification.
- Genome sequencing and analysis, including BLASTn for homology and terminal repeat identification.
Main Results:
- Phage vB_EfaP_IME195, a member of the Podoviridae family, was isolated and characterized.
- Optimal growth conditions yielded a latent period of ~30 min and a burst size of ~120 pfu/cell.
- The phage possesses a linear, double-stranded DNA genome (18,607 bp) with unique 59-bp terminal inverted repeats.
- Genomic analysis revealed 92% identity to the unpublished Enterococcus faecalis phage Idefix.
Conclusions:
- The novel lytic Enterococcus faecalis phage vB_EfaP_IME195 has been successfully isolated and characterized.
- Its unique genomic features, including terminal inverted repeats, offer valuable insights for phage research.
- This phage serves as a foundation for developing phage-based therapies, such as phage cocktails, against multidrug-resistant Enterococcus faecalis.
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