T4-like Escherichia coli phages from the environment carry blaCTX-M

Abstract

The resistance determinant bla<inf>CTX-M</inf> has many variants and has been the most commonly reported gene in clinical isolates of extended spectrum beta-lactamase producing Escherichia coli. Phages have been speculated as potential reservoirs of resistance genes and efficient vehicles for horizontal gene transfer. The objective of the study was to determine the prevalence and characterize bacteriophages that harbour the resistance determinant bla<inf>CTX-M</inf>. Escherichia coli specific bacteriophages were isolated from 15 samples including soil and water across Mangaluru, India using bacterial hosts that were sensitive to ?-lactams. Phenotypic and genotypic characterization based on plaque morphology, host range, restriction fragment length polymorphism (RFLP), presence of bla<inf>CTX-M</inf> and electron microscopy was performed. Of 36 phages isolated, seven were positive for Group 1 of bla<inf>CTX-M</inf>. Based on host range and RFLP pattern, the seven phages were classified into four distinct groups, each harbouring a variant of bla<inf>CTX-M</inf>. Five phages were T4-like Myoviridae by electron microscopy which was further confirmed by polymerase chain reaction (PCR) for T4 specific gp14. Generalized transduction of the CTX-M gene from these phages was also observed. The high prevalence (20%) of this gene bla<inf>CTX-M</inf> in the phage pool confirms the significant role of Myoviridae members, specifically T4-like phages in the dissemination of this resistance gene. Significance and Impact of the Study: The CTX-M gene that confers resistance to Beta-lactam class of drugs is widespread and diverse. Understanding mechanisms of antimicrobial resistance transfer is a key to devise methods for controlling it. Few studies indicate that bacteriophages are involved in the transfer of this gene but the type of phages involved and the degree of involvement remains to be explored. Our work has been able to identify the class of phages and the magnitude of involvement in the dissemination of this gene. © 2018 The Society for Applied Microbiology