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Isolation and Characterization of Lytic Bacteriophages against Antibiotic-Resistant Klebsiella Pneumoniae from Wastewater Samples

Saba M, Farid AJ, Somayeh S, Ali T, Shahab M

Archives of Razi Institute, 2025

Abstract

One of the most important global health problems in the last decade is the increasing prevalence of antibiotic-resistant Klebsiella pneumoniae. Phage therapy is considered a highly efficient alternative to treat antibiotic-resistant bacteria. Therefore, this study aimed isolate lytic phages against clinical strains of antibiotic-resistant Klebsiella pneumoniae from wastewater samples. In this study, different samples were collected, and the presence of phage was confirmed by a double layer test. The protein and genomic structure were verified using SDS- PAGE and RAPD- PCR techniques. Transmission electron microscopy (TEM) was used for the evaluation of morphology. The isolated phage's stability was assessed by subjecting it to different temperature and pH conditions. An adsorption assay test was conducted to determine the duration of phage absorption to corresponding host bacteria. In this study, we isolated three lytic phages (k.8, k.22, and k.34) that target K. pneumoniae. Transmission electron microscopy (TEM) results indicated that all three phages belong to the Siphoviridae family. Additionally, our investigation demonstrated that these phages remain stable within a temperature range of 4°C to 50°C and a pH range of 5.0 to 9.0. The adsorption assay tests showed that the isolated phages were able to attach to their corresponding host bacteria within 6 to 9 minutes. The present study demonstrated that the isolated phages targeting K. pneumoniae exhibited a favorable…

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