Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates

ABSTRACT Lysogenic bacteriophages can integrate their genome into the bacterial chromosome in the form of a prophage and can promote genetic transfer between bacterial strains in vitro. However, the contribution of lysogenic bacteriophages to the incidence of antimicrobial resistance (AMR) in clinic...

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Main Authors: Tony H. Chang, Julie D. Pourtois, Naomi L. Haddock, Daisuke Furukawa, Kate E. Kelly, Derek F. Amanatullah, Elizabeth Burgener, Carlos Milla, Niaz Banaei, Paul L. Bollyky
Format: Article
Language:English
Published: American Society for Microbiology 2025-03-01
Series:mSphere
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Online Access:https://journals.asm.org/doi/10.1128/msphere.00904-24
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author Tony H. Chang
Julie D. Pourtois
Naomi L. Haddock
Daisuke Furukawa
Kate E. Kelly
Derek F. Amanatullah
Elizabeth Burgener
Carlos Milla
Niaz Banaei
Paul L. Bollyky
author_facet Tony H. Chang
Julie D. Pourtois
Naomi L. Haddock
Daisuke Furukawa
Kate E. Kelly
Derek F. Amanatullah
Elizabeth Burgener
Carlos Milla
Niaz Banaei
Paul L. Bollyky
author_sort Tony H. Chang
collection DOAJ
description ABSTRACT Lysogenic bacteriophages can integrate their genome into the bacterial chromosome in the form of a prophage and can promote genetic transfer between bacterial strains in vitro. However, the contribution of lysogenic bacteriophages to the incidence of antimicrobial resistance (AMR) in clinical settings is poorly understood. Here, in a set of 186 clinical isolates of Pseudomonas aeruginosa collected from respiratory cultures from 82 patients with cystic fibrosis, we evaluate the links between prophage counts and both genomic and phenotypic resistance to six anti-pseudomonal antibiotics: tobramycin, colistin, ciprofloxacin, meropenem, aztreonam, and piperacillin–tazobactam. We identified 239 different prophages in total. We find that P. aeruginosa isolates contain on average 3.06 ± 1.84 (SD) predicted prophages. We find no significant association between the number of prophages per isolate and the minimum inhibitory concentration for any of these antibiotics. We then investigate the relationship between particular prophages and AMR. We identify a single lysogenic phage associated with phenotypic resistance to the antibiotic tobramycin and, consistent with this association, we observe that AMR genes associated with resistance to tobramycin are more likely to be found when this prophage is present. However, we find that they are not encoded directly on prophage sequences. Additionally, we identify a single prophage statistically associated with ciprofloxacin resistance but do not identify any genes associated with ciprofloxacin phenotypic resistance. These findings suggest that prophages are only infrequently associated with the AMR genes in clinical isolates of P. aeruginosa.IMPORTANCEAntibiotic-resistant infections of Pseudomonas aeruginosa (Pa), a leading pathogen in patients with cystic fibrosis (CF), are a global health threat. While lysogenic bacteriophages are known to facilitate horizontal gene transfer, their role in promoting antibiotic resistance in clinical settings remains poorly understood. In our analysis of 186 clinical isolates of P. aeruginosa from CF patients, we find that prophage abundance does not predict phenotypic resistance to key antibiotics but that specific prophages are infrequently associated with tobramycin resistance genes. In addition, we do not find antimicrobial resistance (AMR) genes encoded directly on prophages. These results highlight that while phages can be associated with AMR, phage-mediated AMR transfer may be rare in clinical isolates and difficult to identify. This work is important for future efforts on mitigating AMR in CFCF and other vulnerable populations affected by Pa infections and advances our understanding of bacterial-phage dynamics in clinical infections.
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spelling doaj-art-021f33a69f4341b2b6b2e1df03959bc12025-08-20T02:40:33ZengAmerican Society for MicrobiologymSphere2379-50422025-03-0110310.1128/msphere.00904-24Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolatesTony H. Chang0Julie D. Pourtois1Naomi L. Haddock2Daisuke Furukawa3Kate E. Kelly4Derek F. Amanatullah5Elizabeth Burgener6Carlos Milla7Niaz Banaei8Paul L. Bollyky9Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USADivision of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USAUniversity of California San Francisco Medical School, San Francisco, California, USADivision of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USADivision of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USADepartment of Orthopaedic Surgery, Stanford Hospital and Clinics, Redwood City, California, USADivision of Pediatric Pulmonology and Sleep Medicine, Children’s Hospital Los Angeles, Keck School of Medicine at USC, Los Angeles, California, USACystic Fibrosis Clinic, Department of Medicine, Lucille Packard Children’s Hospital, Stanford University School of Medicine, Stanford, California, USADivision of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USADivision of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USAABSTRACT Lysogenic bacteriophages can integrate their genome into the bacterial chromosome in the form of a prophage and can promote genetic transfer between bacterial strains in vitro. However, the contribution of lysogenic bacteriophages to the incidence of antimicrobial resistance (AMR) in clinical settings is poorly understood. Here, in a set of 186 clinical isolates of Pseudomonas aeruginosa collected from respiratory cultures from 82 patients with cystic fibrosis, we evaluate the links between prophage counts and both genomic and phenotypic resistance to six anti-pseudomonal antibiotics: tobramycin, colistin, ciprofloxacin, meropenem, aztreonam, and piperacillin–tazobactam. We identified 239 different prophages in total. We find that P. aeruginosa isolates contain on average 3.06 ± 1.84 (SD) predicted prophages. We find no significant association between the number of prophages per isolate and the minimum inhibitory concentration for any of these antibiotics. We then investigate the relationship between particular prophages and AMR. We identify a single lysogenic phage associated with phenotypic resistance to the antibiotic tobramycin and, consistent with this association, we observe that AMR genes associated with resistance to tobramycin are more likely to be found when this prophage is present. However, we find that they are not encoded directly on prophage sequences. Additionally, we identify a single prophage statistically associated with ciprofloxacin resistance but do not identify any genes associated with ciprofloxacin phenotypic resistance. These findings suggest that prophages are only infrequently associated with the AMR genes in clinical isolates of P. aeruginosa.IMPORTANCEAntibiotic-resistant infections of Pseudomonas aeruginosa (Pa), a leading pathogen in patients with cystic fibrosis (CF), are a global health threat. While lysogenic bacteriophages are known to facilitate horizontal gene transfer, their role in promoting antibiotic resistance in clinical settings remains poorly understood. In our analysis of 186 clinical isolates of P. aeruginosa from CF patients, we find that prophage abundance does not predict phenotypic resistance to key antibiotics but that specific prophages are infrequently associated with tobramycin resistance genes. In addition, we do not find antimicrobial resistance (AMR) genes encoded directly on prophages. These results highlight that while phages can be associated with AMR, phage-mediated AMR transfer may be rare in clinical isolates and difficult to identify. This work is important for future efforts on mitigating AMR in CFCF and other vulnerable populations affected by Pa infections and advances our understanding of bacterial-phage dynamics in clinical infections.https://journals.asm.org/doi/10.1128/msphere.00904-24antimicrobial activitybacteriophage geneticsbacteriophage therapyantimicrobial safety
spellingShingle Tony H. Chang
Julie D. Pourtois
Naomi L. Haddock
Daisuke Furukawa
Kate E. Kelly
Derek F. Amanatullah
Elizabeth Burgener
Carlos Milla
Niaz Banaei
Paul L. Bollyky
Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
mSphere
antimicrobial activity
bacteriophage genetics
bacteriophage therapy
antimicrobial safety
title Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
title_full Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
title_fullStr Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
title_full_unstemmed Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
title_short Prophages are infrequently associated with antibiotic resistance in Pseudomonas aeruginosa clinical isolates
title_sort prophages are infrequently associated with antibiotic resistance in pseudomonas aeruginosa clinical isolates
topic antimicrobial activity
bacteriophage genetics
bacteriophage therapy
antimicrobial safety
url https://journals.asm.org/doi/10.1128/msphere.00904-24
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