Genomic analysis of five new mycobacteriophages belonging to the F, G, K, and P clusters.

Mycobacterial infections are accountable for some of the most challenging diseases to treat in both humans and veterinary medicine, particularly with the increasing prevalence of multidrug-resistant (MDR) Mycobacterium species. Bacteriophages (phages) are being actively investigated as a promising alternative to antibiotic therapy and mycobacteriophages, phages that infect mycobacteria, have recently been demonstrated to be effective candidates in phage therapy. Therefore, discovery and genomic characterisation of diverse phages capable of infecting and killing pathogenic mycobacteria are crucial steps in both monophage therapy and multi-phage therapy, where two or more phages are used in combination. Beyond their therapeutic potential, mycobacteriophages are also employed in the diagnosis, which is particularly challenging since mycobacteria are intracellular pathogens. In this study, we report the isolation and purification of five novel mycobacteriophages using Mycobacterium smegmatis Mc² 155 as the host strain. Their complete genome sequences, annotations, phylogenetic relationships and comparative genome analyses are presented. The isolated phages belong to diverse clusters: F, G, K and P, contributing to the expanding pool of mycobacteriophages.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-025-04426-y.