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

Introduction: 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.Materials and methods: 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 the duration of phage absorption to corresponding host bacteria.Result: In this study, we isolated three lytic phages (k.۸, k.۲۲, and k.۳۴) 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 ۴°C to ۵۰°C and a pH range of ۵.۰ to ۹.۰. The adsorption assay tests showed that the isolated phages were able to attach to their corresponding host bacteria within ۶ to ۹ minutes. Conclusion: The present study demonstrated that the isolated phages targeting K. pneumoniae exhibited a favorable survival rate when subjected to pH and thermal treatments. Furthermore, the isolated phages exhibited a short latent period, indicating rapid adsorption to the host surface. These findings suggest the potential application of these phages in phage therapy. However, further studies in the field of phage therapy are required.

Keywords:

Klebsiella pneumoniae , Lytic phage , Phage therapy , Wastewater