Presenter: Brian Bona
Authors: Brian Bona, Austin Walker, Isaac Miller, Sajid Safiullah, Ruhul Kuddus
Faculty Advisor: Ruhul Kuddus
Institution: Utah Valley University
Background: Bacteriophage therapy offers a hopeful alternative to antibiotic therapy, which has created an increasingly problematic health concern of antibiotic resistance among pathogenic bacteria. Our objective was to isolate bacteriophages capable of effectively compromising pathogenic bacteria highly prone to develop antibiotic resistance (i.e., Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli), and investigate if bacterial pathogens develop resistance to bacteriophages, which would compromise bacteriophages as a therapeutic agent. Materials and Methods: We obtained clinical isolates of P. aeruginosa (27853) and K. pneumoniae (700603) from ATCC (Manassas VA), and isolated E. coli strain p003 from wastewater from Orem wastewater treatment facility (WWTF) using HardyCHROMTM ESBL agar plates (Hardy Diagnostics, Santa Maria CA). We obtained wastewater samples from several local WWTFs, centrifuged (at 2,000xg for 10 minutes), and filtered (using 0.45-micron cartridges), exposed the bacterial strains to the processed samples using standard procedures, and calculated plaque-forming units (pfu) per ml of the water samples. Results: Multiple samples of fresh wastewater collected over three months from Orem WWTF created plaques ranging 104-105 pfu/ml on plates growing P. aeruginosa and K. pneumoniae strains but no plaques on plates growing the E. coli strain. Some of the stored wastewater samples collected from Payson, Provo, Salem, Spanish Fork, and Springville WTTFs and Utah Lake station 1 and UVU duck pond generated plaques on plates growing the K. pneumoniae but not on plates growing P. aeruginosa and E. coli strains. These latter experiments are to be repeated. Phages enriched from plaques generated extensive plaques on P. aeruginosa and K. pneumoniae strains. Conclusions: Our preliminary results indicate that environmental strains of pathogenic bacteria can be more resistant to phage infections than some clinical isolates. Further research is planned to verify these findings and to find out the mechanisms of phage resistance of environmental isolates.