Isolation and characterization of lytic bacteriophages against multidrug resistant Escherichia coli

Juthamas  Chumsen; Juthamas  Chumsen; Parichat  Phumkhachorn; Tarinee  Chaiwong; Viraphong  Lulitanond; Marutpong  Panya

Authors

Juthamas Chumsen College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani 34190, Thailand
Juthamas Chumsen College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani 34190, Thailand
Parichat Phumkhachorn Department of Biological Science, Faculty of Science, Ubon Ratchathani University, Thailand
Tarinee Chaiwong College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani 34190, Thailand
Viraphong Lulitanond Research and Diagnosis Center for Emerging Infectious Diseases, Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
Marutpong Panya College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani 34190, Thailand

Keywords:

bacteriophage, Escherichia coli, lytic bacteriophage, multidrug resistant bacteria, phage, phage therapy

Abstract

The aim of this study was to isolate and characterize Escherichia coli-specific lytic bacteriophages from different wastewater sources. Three bacteriophages, named JC01, JC02, and JC03, were successfully isolated from wastewater samples collected from the wastewater treatment plants of Kudprakhow community, Sanprasitthiprasong Hospital, and Khong Chiam Hospital, respectively. Host range determination revealed that all bacteriophages had only Escherichia coli as a host, indicating a high specific host range property. The inhibition of clinical isolates of multidrug resistant E. coli showed that bacteriophages JC01, JC02, and JC03 inhibited the growth of E. coli at the percentages of 51.7 (138/267), 52.4 (140/267), and 28.5 (76/267), respectively. All bacteriophages were able to tolerate normal saline and distilled water for up to 40 min but were not able to tolerate ethanol and hydrogen peroxide at every time point tested. Heat stability showed that bacteriophage JC01 had the highest resistance at 60°C after 60 min. Meanwhile, bacteriophages JC02 and JC03 showed resistances at 60°C after 45 min. Bacteriophage classification by genome analysis demonstrated that all the bacteriophages were double-stranded DNA viruses. Viral particle morphology observed by transmission electron microscope found that all bacteriophages had the viral particle composed of a head with a hexagonal shape and long tails with contractile. The size from head to tail of all the bacteriophages was approximately 200 nm. Therefore, based on the International Committee on Taxonomy of Viruses (ICTV) classification, bacteriophages JC01, JC02, and JC03 belonged to Family Myoviridae, Order Caudovirales. Therefore, the bacteriophages derived through this study can be further used to study their potential use in advanced research steps, such as in cell cultures and animal models.

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