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Open AccessArticle

Efficacy of Lytic Phage Cocktails on Staphylococcus aureus and Pseudomonas aeruginosa in Mixed-Species Planktonic Cultures and Biofilms

1
College of Science and Engineering, Flinders University, Bedford Park, SA 5042, Australia
2
St Paul’s Hospital Millennium Medical College, Addis Ababa 1271, Ethiopia
3
Infectious Diseases Unit, Queen Elizabeth Hospital, Woodville, SA 5011, Australia
4
Discipline of Medicine, University of Adelaide, Adelaide, SA 5005, Australia
5
AmpliPhi Australia Pty Ltd., Brookvale, NSW 2100, Australia
6
Basil Hetzel Institute for Translational Health Research, School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA 5001, Australia
7
Department of Microbiology and Infectious Diseases, College of Medicine and Public Health, Flinders University, Bedford Park, SA 5042, Australia
*
Author to whom correspondence should be addressed.
Present Address: Phage Consulting, Sydney, NSW 2095, Australia.
Viruses 2020, 12(5), 559; https://doi.org/10.3390/v12050559 (registering DOI)
Received: 24 April 2020 / Revised: 11 May 2020 / Accepted: 13 May 2020 / Published: 18 May 2020
(This article belongs to the Section Bacterial Viruses)
The efficacy of phages in multispecies infections has been poorly examined. The in vitro lytic efficacies of phage cocktails AB-SA01, AB-PA01, which target Staphylococcus aureus and Pseudomonas aeruginosa, respectively, and their combination against their hosts were evaluated in S. aureus and P. aeruginosa mixed-species planktonic and biofilm cultures. Green fluorescent protein (GFP)-labelled P. aeruginosa PAO1 and mCherry-labelled S. aureus KUB7 laboratory strains and clinical isolates were used as target bacteria. During real-time monitoring using fluorescence spectrophotometry, the density of mCherry S. aureus KUB7 and GFP P. aeruginosa PAO1 significantly decreased when treated by their respective phage cocktail, a mixture of phage cocktails, and gentamicin. The decrease in bacterial density measured by relative fluorescence strongly associated with the decline in bacterial cell counts. This microplate-based mixed-species culture treatment monitoring through spectrophotometry combine reproducibility, rapidity, and ease of management. It is amenable to high-throughput screening for phage cocktail efficacy evaluation. Each phage cocktail, the combination of the two phage cocktails, and tetracycline produced significant biofilm biomass reduction in mixed-species biofilms. This study result shows that these phage cocktails lyse their hosts in the presence of non-susceptible bacteria. These data support the use of phage cocktails therapy in infections with multiple bacterial species. View Full-Text
Keywords: phage cocktail therapy; biofilm; planktonic culture; fluorescence; efficacy; mixed-species culture; Staphylococcus aureus; Pseudomonas aeruginosa phage cocktail therapy; biofilm; planktonic culture; fluorescence; efficacy; mixed-species culture; Staphylococcus aureus; Pseudomonas aeruginosa
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MDPI and ACS Style

Kifelew, L.G.; Warner, M.S.; Morales, S.; Thomas, N.; Gordon, D.L.; Mitchell, J.G.; Speck, P.G. Efficacy of Lytic Phage Cocktails on Staphylococcus aureus and Pseudomonas aeruginosa in Mixed-Species Planktonic Cultures and Biofilms. Viruses 2020, 12, 559.

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