Bacteriophages (phages) are viruses that can infect and destroy bacteria. This phenomenon was first observed in the early 1900s, and researchers immediately recognized phages could potentially be used to control bacterial infections. Today, with the pressing health concern of antibiotic resistance, an alternative approach to battle drug resistant bacteria is crucial. Before the therapeutic potential of phages can be realized, it is vital to discover and characterize phages from the environment. The purpose of this study was to isolate and characterize a novel phage that can infect members of the genus Mycobacterium, some of which are notable human pathogens. 

Starting from a soil sample collected on Western Kentucky University’s campus, a phage capable of infecting Mycobacterium smegmatis, a fast-growing nonpathogenic strain, was isolated, grown and purified. Electron microscopy revealed that the phage, named TribleTrouble, is a member of the Siphoviridae because it has the characteristic icosahedral head and non-contractile tail. Restriction analysis of TribleTrouble’s purified genomic DNA suggested it has a high G+C content. To determine TribleTrouble’s relatedness to other phages, we sequenced its entire genome. This analysis revealed it is a member of the K cluster of mycobacteriophages. Interestingly, K cluster phages can infect both fast and slow growing mycobacteria such as M. tuberculosis. TribleTrouble is now part of a national phage collection and is an important new resource for scientists studying phage therapy.