Isolation and characterization of a novel bacteriophage for Pseudomonas aeruginosa

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2023-05

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Pseudomonas aeruginosa is a frequent nosocomial pathogen with emergent strains that display increasing resistance to a wide range of antibiotics including carbapenems. The goal of our study was to isolate and characterize novel bacteriophages with lytic activity against multidrugresistant (MDR) strains of P. aeruginosa. Isolation of wastewater phages was achieved through a series of plaque assays on a control strain (P. aeruginosa BAA-31). The host range of phage isolates was evaluated in vitro with MDR strains from the CDC Antibiotic-Resistance Isolate Bank. To prepare and sequence a library of phage genomes, we used Illumina's TruSeq® Nano DNA Library Prep Kit and MiSeq® system with reagent kit v2. The Center for Phage Technology's Galaxy platform was used for genome processing and annotation. We report the isolation of the novel Pseudomonas phage Jes517. Host range screening revealed 12 of 55 tested strains (21.8%) were at least partially susceptible to Jes517 with 3 showing consistently strong growth reduction in its presence. The ten closest relatives of Jes517 are in the genus Bruynoghevirus with percent identities ranging from 88.7% to 92.8%. Jes517 has a linear dsDNA genome of 45,608 bp encoding 76 open reading frames (ORFs) including three tRNAs. No known integrases, toxins, or resistance factors were identified during gene annotation. Based on its strong growth suppression of MDR P. aeruginosa and predicted lack of virulence genes, phage Jes517 makes a compelling candidate for further investigation and possible therapy.

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