title: In vitro design of a novel lytic bacteriophage cocktail with therapeutic potential against organisms causing diabetic foot infections.
abstract: In patients with diabetes mellitus, foot infections pose a significant risk. These are complex infections commonly caused by Staphylococcus aureus, Pseudomonas aeruginosa and Acinetobacter baumannii, all of which are potentially susceptible to bacteriophages. Here, we characterized five bacteriophages that we had determined previously to have antimicrobial and wound-healing potential in chronic S. aureus, P. aeruginosa and A. baumannii infections. Morphological and genetic features indicated that the bacteriophages were lytic members of the family Myoviridae or Podoviridae and did not harbour any known bacterial virulence genes. Combinations of the bacteriophages had broad host ranges for the different target bacterial species. The activity of the bacteriophages against planktonic cells revealed effective, early killing at 4 h, followed by bacterial regrowth to pre-treatment levels by 24 h. Using metabolic activity as a measure of cell viability within established biofilms, we found significant cell impairment following bacteriophage exposure. Repeated treatment every 4 h caused a further decrease in cell activity. The greatest effects on both planktonic and biofilm cells occurred at a bacteriophage : bacterium input multiplicity of 10. These studies on both planktonic cells and established biofilms allowed us to better evaluate the effects of a high input multiplicity and a multiple-dose treatment protocol, and the findings support further clinical development of bacteriophage therapy.
authors.affiliation: Internal Medicine Department, Santa Marta's Hospital/Central Lisbon Hospital Center, Lisbon, Portugal.|TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: João J
authors.initials: JJ
authors.lastname: Mendes
authors.affiliation: TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Clara
authors.initials: C
authors.lastname: Leandro
authors.affiliation: Interdisciplinary Center of Research in Animal Health, Faculty of Veterinary Medicine of the University of Lisbon, Lisbon, Portugal.
authors.forename: Carla
authors.initials: C
authors.lastname: Mottola
authors.affiliation: TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Raquel
authors.initials: R
authors.lastname: Barbosa
authors.affiliation: TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Filipa A
authors.initials: FA
authors.lastname: Silva
authors.affiliation: Interdisciplinary Center of Research in Animal Health, Faculty of Veterinary Medicine of the University of Lisbon, Lisbon, Portugal.
authors.forename: Manuela
authors.initials: M
authors.lastname: Oliveira
authors.affiliation: Interdisciplinary Center of Research in Animal Health, Faculty of Veterinary Medicine of the University of Lisbon, Lisbon, Portugal.
authors.forename: Cristina L
authors.initials: CL
authors.lastname: Vilela
authors.affiliation: Institute of Microbiology, Faculty of Medicine of the University of Lisbon, Lisbon, Portugal.
authors.forename: José
authors.initials: J
authors.lastname: Melo-Cristino
authors.affiliation: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Górski
authors.affiliation: Department of Microbiology and Immunology, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.|TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Madalena
authors.initials: M
authors.lastname: Pimentel
authors.affiliation: Department of Microbiology and Immunology, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.|TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Carlos
authors.initials: C
authors.lastname: São-José
authors.affiliation: Centro de Investigação Interdisciplinar Egas Moniz (CiiEM), Superior Institute of Health Sciences Egas Moniz, Monte de Caparica, Portugal.|TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Cavaco-Silva
authors.affiliation: TechnoPhage, S.A., Lisbon, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Garcia
mesh_terms: D040981:Acinetobacter baumannii / Q000502:physiology / Q000821:virology*; D001435:Bacteriophages / Q000235:genetics / Q000502:physiology*; D018441:Biofilms / Q000254:growth & development; D017719:Diabetic Foot / Q000382:microbiology*; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000502:physiology / Q000821:virology*; D013211:Staphylococcus aureus / Q000502:physiology / Q000821:virology*