Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro
Many antibiotic-resistant bacteria carry resistance genes on conjugative plasmids that are transferable to commensals and pathogens. We determined the ability of multiple enteric bacteria to acquire and retransfer a broad-host-range plasmid RP4. We used human-derived commensal <i>Escherichia c...
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MDPI AG
2023-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/1/193 |
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| author | Azam A. Sher Mia E. VanAllen Husnain Ahmed Charles Whitehead-Tillery Sonia Rafique Julia A. Bell Lixin Zhang Linda S. Mansfield |
| author_facet | Azam A. Sher Mia E. VanAllen Husnain Ahmed Charles Whitehead-Tillery Sonia Rafique Julia A. Bell Lixin Zhang Linda S. Mansfield |
| author_sort | Azam A. Sher |
| collection | DOAJ |
| description | Many antibiotic-resistant bacteria carry resistance genes on conjugative plasmids that are transferable to commensals and pathogens. We determined the ability of multiple enteric bacteria to acquire and retransfer a broad-host-range plasmid RP4. We used human-derived commensal <i>Escherichia coli</i> LM715-1 carrying a chromosomal red fluorescent protein gene and green fluorescent protein (GFP)-labeled broad-host-range RP4 plasmid with <i>amp</i>R, <i>tet</i>R, and <i>kan</i>R in in vitro matings to rifampicin-resistant recipients, including <i>Escherichia coli</i> MG1655, Dec5α, <i>Vibrio cholerae</i>, <i>Pseudomonas putida</i>, <i>Pseudomonas aeruginosa</i>, <i>Klebsiella pneumoniae</i>, <i>Citrobacter rodentium</i>, and <i>Salmonella</i> Typhimurium. Transconjugants were quantified on selective media and confirmed using fluorescence microscopy and PCR for the GFP gene. The plasmid was transferred from <i>E. coli</i> LM715-1 to all tested recipients except <i>P. aeruginosa</i>. Transfer frequencies differed between specific donor–recipient pairings (10<sup>−2</sup> to 10<sup>−8</sup>). Secondary retransfer of plasmid from transconjugants to <i>E. coli</i> LM715-1 occurred at frequencies from 10<sup>−2</sup> to 10<sup>−7</sup>. A serial passage plasmid persistence assay showed plasmid loss over time in the absence of antibiotics, indicating that the plasmid imposed a fitness cost to its host, although some plasmid-bearing cells persisted for at least ten transfers. Thus, the RP4 plasmid can transfer to multiple clinically relevant bacterial species without antibiotic selection pressure. |
| first_indexed | 2024-03-09T11:38:44Z |
| format | Article |
| id | doaj.art-f2ae78b23af146a3bad323825fe87db8 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T11:38:44Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-f2ae78b23af146a3bad323825fe87db82023-11-30T23:37:46ZengMDPI AGMicroorganisms2076-26072023-01-0111119310.3390/microorganisms11010193Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In VitroAzam A. Sher0Mia E. VanAllen1Husnain Ahmed2Charles Whitehead-Tillery3Sonia Rafique4Julia A. Bell5Lixin Zhang6Linda S. Mansfield7Comparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USADepartment of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USAComparative Enteric Diseases Laboratory, East Lansing, MI 48824, USAMany antibiotic-resistant bacteria carry resistance genes on conjugative plasmids that are transferable to commensals and pathogens. We determined the ability of multiple enteric bacteria to acquire and retransfer a broad-host-range plasmid RP4. We used human-derived commensal <i>Escherichia coli</i> LM715-1 carrying a chromosomal red fluorescent protein gene and green fluorescent protein (GFP)-labeled broad-host-range RP4 plasmid with <i>amp</i>R, <i>tet</i>R, and <i>kan</i>R in in vitro matings to rifampicin-resistant recipients, including <i>Escherichia coli</i> MG1655, Dec5α, <i>Vibrio cholerae</i>, <i>Pseudomonas putida</i>, <i>Pseudomonas aeruginosa</i>, <i>Klebsiella pneumoniae</i>, <i>Citrobacter rodentium</i>, and <i>Salmonella</i> Typhimurium. Transconjugants were quantified on selective media and confirmed using fluorescence microscopy and PCR for the GFP gene. The plasmid was transferred from <i>E. coli</i> LM715-1 to all tested recipients except <i>P. aeruginosa</i>. Transfer frequencies differed between specific donor–recipient pairings (10<sup>−2</sup> to 10<sup>−8</sup>). Secondary retransfer of plasmid from transconjugants to <i>E. coli</i> LM715-1 occurred at frequencies from 10<sup>−2</sup> to 10<sup>−7</sup>. A serial passage plasmid persistence assay showed plasmid loss over time in the absence of antibiotics, indicating that the plasmid imposed a fitness cost to its host, although some plasmid-bearing cells persisted for at least ten transfers. Thus, the RP4 plasmid can transfer to multiple clinically relevant bacterial species without antibiotic selection pressure.https://www.mdpi.com/2076-2607/11/1/193antibiotic resistanceRP4 conjugative plasmidenteric pathogenscommensal bacteria |
| spellingShingle | Azam A. Sher Mia E. VanAllen Husnain Ahmed Charles Whitehead-Tillery Sonia Rafique Julia A. Bell Lixin Zhang Linda S. Mansfield Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro Microorganisms antibiotic resistance RP4 conjugative plasmid enteric pathogens commensal bacteria |
| title | Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro |
| title_full | Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro |
| title_fullStr | Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro |
| title_full_unstemmed | Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro |
| title_short | Conjugative RP4 Plasmid-Mediated Transfer of Antibiotic Resistance Genes to Commensal and Multidrug-Resistant Enteric Bacteria In Vitro |
| title_sort | conjugative rp4 plasmid mediated transfer of antibiotic resistance genes to commensal and multidrug resistant enteric bacteria in vitro |
| topic | antibiotic resistance RP4 conjugative plasmid enteric pathogens commensal bacteria |
| url | https://www.mdpi.com/2076-2607/11/1/193 |
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