Isolation and characterization of novel plasmid-dependent phages infecting bacteria carrying diverse conjugative plasmids

Abstract

Plasmid-dependent phages infect bacteria carrying conjugative plasmids by recognizing the plasmid-encoded pilus. Despite the high abundance of conjugative plasmids in diverse environments, plasmid-dependent phages have not been widely studied. Since conjugative plasmids often carry antimicrobial resistance genes (ARGs), interfering with conjugation could reduce the spread of ARGs and avoid the appearance of multiresistant superbugs. Our aim was to isolate and characterize plasmid-dependent phages able to infect bacteria carrying diverse conjugative plasmids belonging to the most common plasmid families among Gram-negative pathogens. We isolated two lytic phages from wastewater using an avirulent strain of Salmonella enterica carrying the conjugative IncN plasmid pKM101. Both phages, named Lu221 and Hi226, are novel dsDNA viruses within the class Caudoviricetes with genomes of approximately 76 kb. They showed broad host range infecting Escherichia coli , S. enterica , Kluyvera sp., and Enterobacter sp. carrying conjugative plasmids. They recognize plasmid-encoded receptors from 12 out of 15 tested plasmids, all of them carrying resistance determinants. Phages Lu221 and Hi226 could have the potential to help combat the antimicrobial resistance crisis by reducing ARGs present in conjugative plasmids. IMPORTANCE This work was undertaken because plasmid-dependent phages can reduce the prevalence of conjugative plasmids and can be leveraged to prevent the acquisition and dissemination of ARGs by bacteria. The two novel phages described in this study, Lu221 and Hi226, can infect Escherichia coli , Salmonella enterica , Kluyvera sp. and Enterobacter sp. carrying conjugative plasmids. This was verified with plasmids carrying resistance determinants and belonging to the most common plasmid families among Gram-negative pathogens. Therefore, the newly isolated phages could have the potential to help control the spread of ARGs and thus help combat the antimicrobial resistance crisis.