Isolation of Pseudomonas aeruginosa Phages vB_Psu_NEU2023 and vB_Psu_NEU2024 Leads to the Establishment of a New Genus, Alasiavirus

Abstract

Introduction: Pseudomonas aeruginosa is an opportunistic pathogen responsible for severe infections, particularly in immunocompromised patients. The rise of multidrug-resistant P. aeruginosa has become a significant global health concern, highlighting the need for alternative strategies to combat infection. Methods: Two Pseudomonas phages, vB_Psu_NEU2023 and vB_Psu_NEU2024, were isolated from hospital wastewater using plaque assays. Phages were imaged using transmission electron microscopy. DNA was extracted and sequenced using nanopore technology, and bioinformatic analysis uncovered the phage taxonomy. Results: Phages vB_Psu_NEU2023 and vB_Psu_NEU2024 have inovirus and siphovirus morphology, respectively. Whole-genome analysis showed vB_Psu_NEU2023 to belong to a novel genus, here named Alasiavirus, within the Inoviridae family, while vB_Psu_NEU2024 displayed a close relationship to Yuavirus phages within the Mesyanzhinovviridae family. Both phages showed lytic activity against clinical multidrug-resistant P. aeruginosa strains. vB_Psu_NEU2024 genome annotation saw an absence of genes associated with lysogeny and toxin production. Conclusions: This study underscores the importance of hospital wastewater as a valuable reservoir for novel phages with potential clinical applications against multidrug-resistant pathogens.