Metagenomic insights into the global occurrence of ESKAPE-targeting bacteriophages in urban wastewater.

Antimicrobial resistance (AMR) poses a serious threat to global public health, with Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species - collectively known as ESKAPE pathogens - as high priority threats. Bacteriophages represent a potential alternative therapy for AMR infections. While many of the bacteriophages that target ESKAPE pathogens have been isolated from environmental sources, such as wastewater, the global distribution of these bacteriophages remains poorly understood. In this study, metagenomic sequencing of wastewater was used to assess the presence and distribution of known bacteriophages in urban wastewater collected from 62 sites worldwide. Focusing specifically on lytic bacteriophages known to target ESKAPE pathogens, a literature review was conducted to curate a collection of bacteriophages with known therapeutic properties and publicly available genomes. The presence and distribution of these therapeutic bacteriophages in wastewater were examined, both geographically and temporally. Our analysis revealed a high abundance of bacteriophage families associated with the human gastrointestinal tract, as well as those infecting bacteria native to aquatic environments. Bacteriophages similar to those targeting ESKAPE pathogens were detected across the wastewater samples, except for bacteriophages targeting Staphylococcus aureus. Both the presence and abundance of these putative ESKAPE-phages varied considerably across regions, with African sample sites exhibiting the highest diversity and European sample sites the lowest. No clear temporal trends were observed for these bacteriophages. This study provides the use of wastewater to provide insights into the global diversity and distribution of bacteriophages, with a specific focus on species with therapeutic potential against ESKAPE pathogens, while further offering a valuable dataset for future bacteriophage research.