Abstract
The escalating global threat of multidrug-resistant (MDR) Pseudomonas aeruginosa, designated by the WHO as a critical-priority pathogen, necessitates urgent development of alternative therapeutics. We present Pseudomonas phage Arefeen1, isolated from Bangladeshi wastewater, as a clinically translatable candidate with therapeutic potential. Comprehensive genomic characterization revealed a 46,021 bp strictly lytic genome (52.68% GC content) belonging to the Caudoviricetes class, completely lacking virulence factors or antibiotic resistance genes as confirmed by VFDB and ResFinder screening-a crucial safety profile for clinical application. Phenotypically, Arefeen1 demonstrated efficient in vitro lytic activity, exhibiting rapid replication kinetics (25-min latent period), high burst size (150 ± 12 PFU/cell), and robust production yields (≥ 10(9) PFU/mL post-PEG precipitation). Host range analysis showed 85.7% efficacy against a panel of clinically relevant MDR strains, including respiratory (PA_CU_1) and wound (MO_4642.012.002) isolates. Comparative genomics with 840 P. aeruginosa phages identified six unique genes encoding membrane-interaction proteins (CDS_0052/0058/0061) and a specialized tRNA complement, suggesting evolutionary adaptations for enhanced host range and translational efficiency. Its isolation from Bangladesh's unique microbial ecosystem provides a geographically optimized resource for LMICs disproportionately affected by antimicrobial resistance. These findings, combined with its clinical strain coverage, position Arefeen1 as a potential candidate for preclinical development and phage therapy implementation against this formidable pathogen.