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Characterization of wastewater-derived bacteriophages infecting Enterococcus faecalis in Bulgaria: insights into the

Yoana Kizheva1, Tsveta Dimova1, Maria Pandova1

  • 1Department of General and Industrial Microbiology, Faculty of Biology, Sofia University "St. Kliment Ohridski", Sofia, Bulgaria.

Frontiers in Microbiology
|December 29, 2025
PubMed

Insights

New bacteriophages were discovered that can eliminate *Enterococcus faecalis*, including antibiotic-resistant strains. Phage vB_SEF_8 shows promise as a therapeutic agent against this opportunistic pathogen.

Area of Science:

  • Microbiology
  • Virology
  • Bacteriology

Background:

  • *Enterococcus faecalis* is an opportunistic pathogen causing nosocomial infections and posing treatment challenges due to genetic adaptability.
  • Antibiotic resistance and virulence gene acquisition in *E. faecalis* necessitate novel therapeutic strategies like phage therapy.
  • Wastewater environments are potential sources for discovering effective bacteriophages.

Purpose of the Study:

  • To isolate and characterize bacteriophages targeting *Enterococcus faecalis* from wastewater.
  • To evaluate the biological properties and lytic potential of isolated phages against *E. faecalis*.
  • To assess the therapeutic potential of the most promising phage isolate.

Main Methods:

  • Isolation of bacteriophages from wastewater samples.
  • Host range determination against *E. faecalis* and other *Enterococcus* species.
  • Stability assays (pH, temperature), RFLP analysis, TEM, DNA sequencing, and phylogenetic analysis.
  • Evaluation of phage efficacy in a milk-based matrix.

Main Results:

  • Three bacteriophages (vB_SEF_8, vB_SEF_13, vB_SEF_15) were isolated, exclusively targeting *E. faecalis*, including multidrug-resistant strains.
  • Phages exhibited broad pH and temperature stability, with long-term viability at 4°C.
  • Phage vB_SEF_8, a novel species within *Saphexavirus*, demonstrated broad host range, stability, and successful inhibition of *E. faecalis* in a milk matrix.

Conclusions:

  • The isolated bacteriophages, particularly vB_SEF_8, are effective against *E. faecalis*.
  • vB_SEF_8 possesses desirable characteristics for a therapeutic agent, including stability and lack of virulence/resistance genes.
  • Phage therapy presents a viable alternative for controlling *E. faecalis* infections and contamination.
Keywords:
Enterococcus faecalisSaphexavirus phagesantibiotic resistancefood safetyphage therapywastewater

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