Isolation and characterization of novel phage (Podoviridae ɸParuNE1) and its efficacy against multi-drug-resistant Pseudomonas aeruginosa planktonic cells and biofilm
Nkechi V. Enwuru, Jason J. Gill, Katri P. Anttonen, Christian A. Enwuru, Ry Young, Akinloye O. Coker, Jeffrey D. Cirillo
Abstract
Abstract Background Bacterial pathogen ( Pseudomonas aeruginosa ) could form biofilm that conveys multi-drug resistance. Bacteriophage as an alternative to antibacterial resistance is useful against biofilm complications. This study evaluated antibacterial and biofilm removal activities of lytic phage, specific against multi-drug-resistant clinical P. aeruginosa. Results The phage showed a wide range of pH (5–10) and heat (7–44 °C) stability. Electron microscopy showed ɸPauNE1 phage head (60 nm in diameter) and non-contractile tail (12 nm in length by 8 nm in width); hence, the family Podoviridae and the order Caudovirales . Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed structured protein of 55 kDa and double-stranded DNA of 45 kb. The phage was species specific and had broad host range activity. It inhibited bacterial growth at multiplicity of infection (MOI) 1–0.000001 pfu/ml. Inhibition was maximal at both low (1 × 10 5 ) and high (1 × 10 9 ) bacterial CFU/ml. Biofilm removal test showed that the phage removed more than 60% cell biomass within CFU/ml of 1.5 × 10 8 , 6.0 × 10 8 and l.0 × 10 9 . Conclusion Phage (ɸPauNE1) was unique and had broad host range activity. The phage exhibited strong bacteriolytic activity against biofilm forming multi-drug-resistant strains. It had no lytic effect on the heterogeneous strains and so a promising bioagent.