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Updated: Jun 12, 2025

Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes
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Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes

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Systematic bacteriophage selection for the lysis of multiple Pseudomonas aeruginosa strains

Finja Rieper1,2, Johannes Wittmann3, Boyke Bunk3

  • 1Pharmaceutical Biotechnology, Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Braunschweig, Germany.

Frontiers in Cellular and Infection Microbiology
|June 9, 2025
PubMed

Insights

This study explored bacteriophages to treat antibiotic-resistant Pseudomonas aeruginosa infections. Researchers identified effective phage combinations, laying the groundwork for developing broad-spectrum phage therapy against this pathogen.

Area of Science:

  • Microbiology
  • Infectious Diseases
  • Biotechnology

Background:

  • Pseudomonas aeruginosa is a highly antibiotic-resistant opportunistic pathogen causing severe infections.
  • Phage therapy, using P. aeruginosa-specific bacteriophages, presents a promising alternative to antibiotics.
  • Effective phage therapy requires careful selection, production, and administration of multiple phage strains.

Purpose of the Study:

  • To evaluate the efficacy of 25 bacteriophages against 141 clinical isolates of Pseudomonas aeruginosa.
  • To compare host spectrum analyses using double agar overlay plaque assay (DPA) and planktonic killing assay (PKA).
  • To identify potent phage combinations for broad-spectrum therapeutic application against P. aeruginosa.

Main Methods:

  • Susceptibility testing of 25 phages against 141 P. aeruginosa strains using DPA and PKA.
  • Classification of phages into known genera and identification of potential new species.
  • Exclusion of phages with lysogenic potential.
  • In silico analysis of phage combinations for synergistic efficacy.

Main Results:

  • DPA and PKA yielded concordant host range results in 70% of cases, with PKA showing higher potential for diagnostics due to automation.
  • Several phage genera, including Pakpunavirus, Pbunavirus, Pawinskivirus, Elvirus, Litunavirus, and Bruynoghevirus, demonstrated significant lytic activity.
  • Seven phages were disqualified due to the presence of genetic determinants for lysogenicity.
  • In silico testing identified highly effective phage combinations capable of eradicating multiple P. aeruginosa strains.

Conclusions:

  • Bacteriophage therapy is a viable strategy for combating antibiotic-resistant Pseudomonas aeruginosa.
  • Planktonic killing assay (PKA) is a suitable high-throughput method for phage diagnostics.
  • The identified phage combinations provide a foundation for developing a broad-spectrum phage therapy against P. aeruginosa.
Keywords:
Pseudomonas aeruginosaantibiotic resistancebacteriophagesphage selectionphage susceptibility testing

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