| Summary: | Bacteriophages could be a useful adjunct to antibiotics for the treatment of multidrug-resistant <i>Pseudomonas aeruginosa</i> infections. In this study, lytic <i>P. aeruginosa</i> myoviruses PsCh, PsIn, Ps25, and Ps12on-D were isolated from Tunisian sewage samples. Phage Ps12on-D displayed an adsorption time of ~10 min, a short latency period (~10 min), and a large burst size (~115 PFU per infected cell) under standard growth conditions. All phages were active at broad temperature (4 °C to 50 °C) and pH (3.0 to 11.0) ranges and were able to lyse a wide variety of <i>P. aeruginosa</i> strains isolated from clinical and environmental samples worldwide. Illumina sequencing revealed double-stranded DNA genomes ranging from 87,887 and 92,710 bp with high sequence identity to <i>Pseudomonas</i> phage PAK_P1. All four phages based on sequence analysis were assigned to the <i>Pakpunavirus</i> genus. The presented characterization and preclinical assessment are part of an effort to establish phage therapy treatment as an alternative strategy for the management of multidrug-resistant <i>P. aeruginosa</i> infections in Tunisia.
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