Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 13 2 2021 04 10 225 234 Fatemeh Habibinava Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran Mohammad Reza Zolfaghari Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran Mohsen Zargar Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran Salehe Sabouri Shahrbabak Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran; Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran Mohammad Soleimani Department of Microbiology, School of Medicine, AJA University of Medical Sciences, Tehran, Iran; Infectious Diseases Research Center, AJA University of Medical Sciences, Tehran, Iran 2020 10 06 2021 03 20 Background and Objectives: Multi-drug-resistant Enterobacter aerogenes is associated with various infectious diseases that cannot be easily treated by antibiotics. However, bacteriophages have potential therapeutic applications in the control of multi-drug-resistant bacteria. In this study, we aimed to isolate and characterize of a lytic bacteriophage that can lyse specifically the multi-drug-resistant (MDR) E. aerogenes. Materials and Methods: Lytic bacteriophage was isolated from Qaem hospital wastewater and characterized morphologically and genetically. Next-generation sequencing was used to complete genome analysis of the isolated bacteriophage. Results: Based on the transmission electron microscopy feature, the isolated bacteriophage (vB-Ea-5) belongs to the family Myoviridae. vB-Ea-5 had a latent period of 25 minutes, a burst size of 13 PFU/ml, and a burst time of 40 min. Genome sequencing revealed that vB-Ea-5 has a 135324 bp genome with 41.41% GC content. The vB-Ea-5 genome codes 212 ORFs 90 of which were categorized into several functional classes such as DNA replication and modification, transcriptional regulation, packaging, structural proteins, and a host lysis protein (Holin). No antibiotic resistance and toxin genes were detected in the genome. SDS-PAGE of vB-Ea-5 proteins exhibited three major and four minor bands with a molecular weight ranging from 18 to 50 kD. Conclusion: Our study suggests vB-Ea-5 as a potential candidate for phage therapy against MDR E. aerogenes infections. https://ijm.tums.ac.ir/index.php/ijm/article/view/2803 https://ijm.tums.ac.ir/index.php/ijm/article/download/2803/1337