Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli

Mehreen Anjum, Jonas Stenløkke Madsen, Joseph Nesme, Bimal Jana, Maria Wiese, Džiuginta Jasinskytė, Dennis Sandris Nielsen, Søren Johannes Sørensen, Anders Dalsgaard, Arshnee Moodley, Valeria Bortolaia, Luca Guardabassi*

*Corresponding author for this work

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Abstract

The gut is a hot spot for transfer of antibiotic resistance genes from ingested exogenous bacteria to the indigenous microbiota. The objective of this study was to determine the fate of two nearly identical bla CMY-2-harboring plasmids introduced into the human fecal microbiota by two Escherichia coli strains isolated from human and poultry meat, respectively. The chromosome and the CMY-2-encoding plasmid of both strains were labeled with distinct fluorescent markers (mCherry and GFP), allowing Fluorescence Activated Cell Sorting (FACS)-based tracking of the strain and the resident bacteria that have acquired its plasmid. Each strain was introduced into an established in vitro gut model (CoMiniGut) inoculated with individual feces from ten healthy volunteers. Fecal samples collected 2, 6 and 24 h after strain inoculation were analyzed by FACS and plate counts. Although the human strain survived better than the poultry meat strain, both strains transferred their plasmids to the fecal microbiota at concentrations as low as 102 CFU/mL. Strain survival and plasmid transfer varied significantly depending on inoculum concentration and individual fecal microbiota. Identification of transconjugants by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry revealed that the plasmids were predominantly acquired by Enterobacteriaceae such as E. coli and Hafnia alvei. Our experimental data demonstrate that exogenous E. coli of human or animal origin can readily transfer CMY-2-encoding IncI1 plasmids to the human fecal microbiota. Low amounts of exogenous strain are sufficient to ensure plasmid transfer if the strain is able to survive the gastric environment.
Original languageEnglish
Article numbere02528
JournalAntimicrobial Agents and Chemotherapy
Volume63
Issue number5
Number of pages13
ISSN0066-4804
DOIs
Publication statusPublished - 2019

Keywords

  • CoMiniGut model
  • Escherichia coli
  • Incl1
  • Cephalosporin

Cite this

Anjum, M., Madsen, J. S., Nesme, J., Jana, B., Wiese, M., Jasinskytė, D., ... Guardabassi, L. (2019). Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli. Antimicrobial Agents and Chemotherapy, 63(5), [e02528]. https://doi.org/10.1128/AAC.02528-18
Anjum, Mehreen ; Madsen, Jonas Stenløkke ; Nesme, Joseph ; Jana, Bimal ; Wiese, Maria ; Jasinskytė, Džiuginta ; Nielsen, Dennis Sandris ; Sørensen, Søren Johannes ; Dalsgaard, Anders ; Moodley, Arshnee ; Bortolaia, Valeria ; Guardabassi, Luca. / Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli. In: Antimicrobial Agents and Chemotherapy. 2019 ; Vol. 63, No. 5.
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title = "Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli",
abstract = "The gut is a hot spot for transfer of antibiotic resistance genes from ingested exogenous bacteria to the indigenous microbiota. The objective of this study was to determine the fate of two nearly identical bla CMY-2-harboring plasmids introduced into the human fecal microbiota by two Escherichia coli strains isolated from human and poultry meat, respectively. The chromosome and the CMY-2-encoding plasmid of both strains were labeled with distinct fluorescent markers (mCherry and GFP), allowing Fluorescence Activated Cell Sorting (FACS)-based tracking of the strain and the resident bacteria that have acquired its plasmid. Each strain was introduced into an established in vitro gut model (CoMiniGut) inoculated with individual feces from ten healthy volunteers. Fecal samples collected 2, 6 and 24 h after strain inoculation were analyzed by FACS and plate counts. Although the human strain survived better than the poultry meat strain, both strains transferred their plasmids to the fecal microbiota at concentrations as low as 102 CFU/mL. Strain survival and plasmid transfer varied significantly depending on inoculum concentration and individual fecal microbiota. Identification of transconjugants by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry revealed that the plasmids were predominantly acquired by Enterobacteriaceae such as E. coli and Hafnia alvei. Our experimental data demonstrate that exogenous E. coli of human or animal origin can readily transfer CMY-2-encoding IncI1 plasmids to the human fecal microbiota. Low amounts of exogenous strain are sufficient to ensure plasmid transfer if the strain is able to survive the gastric environment.",
keywords = "CoMiniGut model, Escherichia coli, Incl1, Cephalosporin",
author = "Mehreen Anjum and Madsen, {Jonas Stenl{\o}kke} and Joseph Nesme and Bimal Jana and Maria Wiese and Džiuginta Jasinskytė and Nielsen, {Dennis Sandris} and S{\o}rensen, {S{\o}ren Johannes} and Anders Dalsgaard and Arshnee Moodley and Valeria Bortolaia and Luca Guardabassi",
year = "2019",
doi = "10.1128/AAC.02528-18",
language = "English",
volume = "63",
journal = "Antimicrobial Agents and Chemotherapy",
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publisher = "American Society for Microbiology",
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Anjum, M, Madsen, JS, Nesme, J, Jana, B, Wiese, M, Jasinskytė, D, Nielsen, DS, Sørensen, SJ, Dalsgaard, A, Moodley, A, Bortolaia, V & Guardabassi, L 2019, 'Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli', Antimicrobial Agents and Chemotherapy, vol. 63, no. 5, e02528. https://doi.org/10.1128/AAC.02528-18

Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli. / Anjum, Mehreen; Madsen, Jonas Stenløkke; Nesme, Joseph; Jana, Bimal; Wiese, Maria; Jasinskytė, Džiuginta; Nielsen, Dennis Sandris; Sørensen, Søren Johannes; Dalsgaard, Anders; Moodley, Arshnee; Bortolaia, Valeria; Guardabassi, Luca.

In: Antimicrobial Agents and Chemotherapy, Vol. 63, No. 5, e02528, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Fate of CMY-2-encoding plasmids introduced into the human fecal microbiota by exogenous Escherichia coli

AU - Anjum, Mehreen

AU - Madsen, Jonas Stenløkke

AU - Nesme, Joseph

AU - Jana, Bimal

AU - Wiese, Maria

AU - Jasinskytė, Džiuginta

AU - Nielsen, Dennis Sandris

AU - Sørensen, Søren Johannes

AU - Dalsgaard, Anders

AU - Moodley, Arshnee

AU - Bortolaia, Valeria

AU - Guardabassi, Luca

PY - 2019

Y1 - 2019

N2 - The gut is a hot spot for transfer of antibiotic resistance genes from ingested exogenous bacteria to the indigenous microbiota. The objective of this study was to determine the fate of two nearly identical bla CMY-2-harboring plasmids introduced into the human fecal microbiota by two Escherichia coli strains isolated from human and poultry meat, respectively. The chromosome and the CMY-2-encoding plasmid of both strains were labeled with distinct fluorescent markers (mCherry and GFP), allowing Fluorescence Activated Cell Sorting (FACS)-based tracking of the strain and the resident bacteria that have acquired its plasmid. Each strain was introduced into an established in vitro gut model (CoMiniGut) inoculated with individual feces from ten healthy volunteers. Fecal samples collected 2, 6 and 24 h after strain inoculation were analyzed by FACS and plate counts. Although the human strain survived better than the poultry meat strain, both strains transferred their plasmids to the fecal microbiota at concentrations as low as 102 CFU/mL. Strain survival and plasmid transfer varied significantly depending on inoculum concentration and individual fecal microbiota. Identification of transconjugants by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry revealed that the plasmids were predominantly acquired by Enterobacteriaceae such as E. coli and Hafnia alvei. Our experimental data demonstrate that exogenous E. coli of human or animal origin can readily transfer CMY-2-encoding IncI1 plasmids to the human fecal microbiota. Low amounts of exogenous strain are sufficient to ensure plasmid transfer if the strain is able to survive the gastric environment.

AB - The gut is a hot spot for transfer of antibiotic resistance genes from ingested exogenous bacteria to the indigenous microbiota. The objective of this study was to determine the fate of two nearly identical bla CMY-2-harboring plasmids introduced into the human fecal microbiota by two Escherichia coli strains isolated from human and poultry meat, respectively. The chromosome and the CMY-2-encoding plasmid of both strains were labeled with distinct fluorescent markers (mCherry and GFP), allowing Fluorescence Activated Cell Sorting (FACS)-based tracking of the strain and the resident bacteria that have acquired its plasmid. Each strain was introduced into an established in vitro gut model (CoMiniGut) inoculated with individual feces from ten healthy volunteers. Fecal samples collected 2, 6 and 24 h after strain inoculation were analyzed by FACS and plate counts. Although the human strain survived better than the poultry meat strain, both strains transferred their plasmids to the fecal microbiota at concentrations as low as 102 CFU/mL. Strain survival and plasmid transfer varied significantly depending on inoculum concentration and individual fecal microbiota. Identification of transconjugants by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry revealed that the plasmids were predominantly acquired by Enterobacteriaceae such as E. coli and Hafnia alvei. Our experimental data demonstrate that exogenous E. coli of human or animal origin can readily transfer CMY-2-encoding IncI1 plasmids to the human fecal microbiota. Low amounts of exogenous strain are sufficient to ensure plasmid transfer if the strain is able to survive the gastric environment.

KW - CoMiniGut model

KW - Escherichia coli

KW - Incl1

KW - Cephalosporin

U2 - 10.1128/AAC.02528-18

DO - 10.1128/AAC.02528-18

M3 - Journal article

VL - 63

JO - Antimicrobial Agents and Chemotherapy

JF - Antimicrobial Agents and Chemotherapy

SN - 0066-4804

IS - 5

M1 - e02528

ER -