Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica
Determinants of multidrug resistance (MDR) are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary heal...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-08-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01459/full |
| _version_ | 1819212775192264704 |
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| author | Maria Hoffmann James B. Pettengill Narjol Gonzalez-Escalona John Miller John Miller Sherry L. Ayers Shaohua Zhao Marc W. Allard Patrick F. McDermott Eric W. Brown Steven R. Monday |
| author_facet | Maria Hoffmann James B. Pettengill Narjol Gonzalez-Escalona John Miller John Miller Sherry L. Ayers Shaohua Zhao Marc W. Allard Patrick F. McDermott Eric W. Brown Steven R. Monday |
| author_sort | Maria Hoffmann |
| collection | DOAJ |
| description | Determinants of multidrug resistance (MDR) are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary health. Our understanding of MDR among Salmonella has been limited by the lack of closed plasmid genomes for comparisons across resistance phenotypes, due to difficulties in effectively separating the DNA of these high-molecular weight, low-copy-number plasmids from chromosomal DNA. To resolve this problem, we demonstrate an efficient protocol for isolating, sequencing and closing IncA/C plasmids from Salmonella sp. using single molecule real-time sequencing on a Pacific Biosciences (Pacbio) RS II Sequencer. We obtained six Salmonella enterica isolates from poultry, representing six different serovars, each exhibiting the MDR-Ampc resistance profile. Salmonella plasmids were obtained using a modified mini preparation and transformed with Escherichia coli DH10Br. A Qiagen Large-Construct kit™ was used to recover highly concentrated and purified plasmid DNA that was sequenced using PacBio technology. These six closed IncA/C plasmids ranged in size from 104 to 191 kb and shared a stable, conserved backbone containing 98 core genes, with only six differences among those core genes. The plasmids encoded a number of antimicrobial resistance genes, including those for quaternary ammonium compounds and mercury. We then compared our six IncA/C plasmid sequences: first with 14 IncA/C plasmids derived from S. enterica available at the National Center for Biotechnology Information (NCBI), and then with an additional 38 IncA/C plasmids derived from different taxa. These comparisons allowed us to build an evolutionary picture of how antimicrobial resistance may be mediated by this common plasmid backbone. Our project provides detailed genetic information about resistance genes in plasmids, advances in plasmid sequencing, and phylogenetic analyses, and important insights about how MDR evolution occurs across diverse serotypes from different animal sources, particularly in agricultural settings where antimicrobial drug use practices vary. |
| first_indexed | 2024-12-23T06:48:19Z |
| format | Article |
| id | doaj.art-671d2452ffda4086859d10d36b25656f |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-23T06:48:19Z |
| publishDate | 2017-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-671d2452ffda4086859d10d36b25656f2022-12-21T17:56:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-08-01810.3389/fmicb.2017.01459288800Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella entericaMaria Hoffmann0James B. Pettengill1Narjol Gonzalez-Escalona2John Miller3John Miller4Sherry L. Ayers5Shaohua Zhao6Marc W. Allard7Patrick F. McDermott8Eric W. Brown9Steven R. Monday10Division of Microbiology, Office of Regulatory Science, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDivision of Public Health Informatics and Analytics, Office of Food Defense, Communication and Emergency Response, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDivision of Microbiology, Office of Regulatory Science, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDivision of Public Health Informatics and Analytics, Office of Food Defense, Communication and Emergency Response, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesU.S. Department of Energy, Oak Ridge Institute for Science and EducationOak Ridge, TN, United StatesDivision of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug AdministrationLaurel, MD, United StatesDivision of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug AdministrationLaurel, MD, United StatesDivision of Microbiology, Office of Regulatory Science, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDivision of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug AdministrationLaurel, MD, United StatesDivision of Microbiology, Office of Regulatory Science, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDivision of Microbiology, Office of Regulatory Science, Center for Food Safety and Nutrition, U.S. Food and Drug AdministrationCollege Park, MD, United StatesDeterminants of multidrug resistance (MDR) are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary health. Our understanding of MDR among Salmonella has been limited by the lack of closed plasmid genomes for comparisons across resistance phenotypes, due to difficulties in effectively separating the DNA of these high-molecular weight, low-copy-number plasmids from chromosomal DNA. To resolve this problem, we demonstrate an efficient protocol for isolating, sequencing and closing IncA/C plasmids from Salmonella sp. using single molecule real-time sequencing on a Pacific Biosciences (Pacbio) RS II Sequencer. We obtained six Salmonella enterica isolates from poultry, representing six different serovars, each exhibiting the MDR-Ampc resistance profile. Salmonella plasmids were obtained using a modified mini preparation and transformed with Escherichia coli DH10Br. A Qiagen Large-Construct kit™ was used to recover highly concentrated and purified plasmid DNA that was sequenced using PacBio technology. These six closed IncA/C plasmids ranged in size from 104 to 191 kb and shared a stable, conserved backbone containing 98 core genes, with only six differences among those core genes. The plasmids encoded a number of antimicrobial resistance genes, including those for quaternary ammonium compounds and mercury. We then compared our six IncA/C plasmid sequences: first with 14 IncA/C plasmids derived from S. enterica available at the National Center for Biotechnology Information (NCBI), and then with an additional 38 IncA/C plasmids derived from different taxa. These comparisons allowed us to build an evolutionary picture of how antimicrobial resistance may be mediated by this common plasmid backbone. Our project provides detailed genetic information about resistance genes in plasmids, advances in plasmid sequencing, and phylogenetic analyses, and important insights about how MDR evolution occurs across diverse serotypes from different animal sources, particularly in agricultural settings where antimicrobial drug use practices vary.http://journal.frontiersin.org/article/10.3389/fmicb.2017.01459/fullantimicrobial resistanceIncA/C plasmidSalmonella entericanext generation sequencing |
| spellingShingle | Maria Hoffmann James B. Pettengill Narjol Gonzalez-Escalona John Miller John Miller Sherry L. Ayers Shaohua Zhao Marc W. Allard Patrick F. McDermott Eric W. Brown Steven R. Monday Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica Frontiers in Microbiology antimicrobial resistance IncA/C plasmid Salmonella enterica next generation sequencing |
| title | Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica |
| title_full | Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica |
| title_fullStr | Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica |
| title_full_unstemmed | Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica |
| title_short | Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica |
| title_sort | comparative sequence analysis of multidrug resistant inca c plasmids from salmonella enterica |
| topic | antimicrobial resistance IncA/C plasmid Salmonella enterica next generation sequencing |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01459/full |
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