Isolation, characterization, and genomic analysis of a novel phage WSPA with lytic activity against .

is an established pathogen implicated in hospital-acquired infections and is notorious for its propensity to form biofilms on medical devices, leading to persistent environmental contamination and increased infection risks. Addressing this challenge, our study introduces a novel bacteriophage, Weishan phage (WSPA), isolated from the gut of L., exhibiting potent activity against multidrug-resistant strains of . Through comprehensive genomic and proteomic characterization, we classified phage WSPA as a member of the genus within the class . The WSPA phage has a double-stranded DNA genome of 173,655 base pairs and a GC content of 40.09%. Of the 273 open reading frames identified, 124 encode for proteins with recognized functions in the National Center for Biotechnology Information (NCBI) database, while the remaining 149 are of unknown function. Additionally, we identified six tRNA genes and did not identify any virulence or antibiotic resistance genes. However, given the presence of numerous hypothetical genes with unknown functions, this phage may possess certain therapeutic safety potential, though additional research validation is still required. Comparative genomic analysis revealed that WSPA shares 86.92% sequence identity with the known (MW082584.1). We further assessed the phage's one-step growth characteristics, thermal and pH stability, and determined its host range, which are critical for its application in environmental and clinical settings. Our findings suggest that bacteriophage WSPA could serve as an eco-friendly and effective agent in controlling infections, with promising implications for phage therapy and biocontrol in healthcare environments.IMPORTANCEThis study isolated a novel phage, WSPA, from L. gut that specifically targets multidrug-resistant . Genomic analysis identified WSPA as a new Muldoonvirus member lacking virulence/resistance genes. With excellent stability and lytic activity, WSPA shows potential for hospital infection control. As the first phage isolated from the cockroach gut, this work expands phage resources and supports medicinal insect phage library development, advancing phage therapy and biocontrol applications.