Role of Phage–Antibiotic Combinations in Reducing ESBL-Producing and Carbapenem-ResistantEscherichia coli

Abstract

ABSTRACTThe emergence of extended-spectrumβ-lactamase (ESBL)-producingE. coliand carbapenem-resistantE. coli(CREC) poses a significant global health concern. Here, we isolated and characterized two novel phages and studied their effectiveness with antibiotics against ESBL-producingE. coliand CREC. The isolated phages, EC.W1-9 and EC.W15-4, belonged to thePodoviridaeandMyoviridaefamilies, respectively. They are safe for bacterial control as they do not contain integrase or toxin-coding genes. The phage combination considerably enhanced lytic ability, effectively lysing 61.7% of the 60E. coliisolates, compared to lysis in the 41.6% –55% range by individual phages. Phages EC.W1-9 and EC.W15-4 combined demonstrated 100% susceptibility against differentE. colisequence types, including ST73, ST648, ST2311, ST405, ST7962, ST131, ST13003, and ST167. Additionally, studies showed synergy between antibiotics and phage combinations against ESBL-producingE.coli, with susceptibility of 73.3% and 54% for CREC. The combined treatment of isolated phages and antibiotics significantly increased survival rates in BALB/c mice exposed to various ST types of ESBL-producingE. coliand CREC, including ST131, ST648, and ST410. Survival rates against KBN7288 (ST131) increased by approximately 75% and 50% compared to individual phages EC.W1-9 and EC.W15-4, respectively. When phages and antibiotics were combined, survival rates againstE. coliisolates KBN5617 (ST410), KBN6241 (ST410), and KBN4004 (ST648) ranged from 75% – 100%. Finally, this study highlights the importance of phage and phage-antibiotic combinations to prepare phages for killing different ST types of ESBL-producingE. coliand CREC isolates.IMPORTANCEWhen combined with antibiotics, phage therapy shows promise in fighting multidrug-resistant bacteria. However, antagonism between phages and antibiotics has been reported. This research isolates and characterizes two novel phages, EC.W1-9 and EC.W15-4, from thePodoviridaeandMyoviridaefamilies, respectively, and evaluates their effectiveness against ESBL-producingE. coliand CREC. These phages, lacking integrase or toxin-coding genes, showed significant promise in bacterial control. Combined phage treatment lysed 61.7% ofE.coliisolates, outperforming individual phages. The phage combination showed 100% susceptibility against differentE. colisequence types. Additionally, the synergy between phages and antibiotics increased susceptibility rates to 73.3% for ESBL-producingE. coliand 54% for CREC. In BALB/c mice, combined treatments significantly improved survival rates against variousE. coliisolates. Finally. this study emphasizes the potential of phage and phage-antibiotic combinations in targeting various ST types of ESBL-producingE. coliand CREC.