Conjugative transfer of multi-drug resistance IncN plasmids from environmental waterborne bacteria to Escherichia coli

Jessica Guzman-Otazo, Enrique Joffré, Jorge Agramont, Nataniel Mamani, Jekaterina Jutkina, Fredrik Boulund, Yue O.O. Hu, Daphne Jumilla-Lorenz, Anne Farewell, D. G.Joakim Larsson, Carl Fredrik Flach, Volga Iñiguez, Åsa Sjöling

Research output: Contribution to journalArticlepeer-review

Abstract

Watersheds contaminated with municipal, hospital, and agricultural residues are recognized as reservoirs for bacteria carrying antibiotic resistance genes (ARGs). The objective of this study was to determine the potential of environmental bacterial communities from the highly contaminated La Paz River basin in Bolivia to transfer ARGs to an Escherichia coli lab strain used as the recipient. Additionally, we tested ZnSO4 and CuSO4 at sub-inhibitory concentrations as stressors and analyzed transfer frequencies (TFs), diversity, richness, and acquired resistance profiles. The bacterial communities were collected from surface water in an urban site close to a hospital and near an agricultural area. High transfer potentials of a large set of resistance factors to E. coli were observed at both sites. Whole-genome sequencing revealed that putative plasmids belonging to the incompatibility group N (IncN, IncN2, and IncN3) were predominant among the transconjugants. All IncN variants were verified to be mobile by a second conjugation step. The plasmid backbones were similar to other IncN plasmids isolated worldwide and carried a wide range of ARGs extensively corroborated by phenotypic resistance patterns. Interestingly, all transconjugants also acquired the class 1 integron intl1, which is commonly known as a proxy for anthropogenic pollution. The addition of ZnSO4 and CuSO4 at sub-inhibitory concentrations did not affect the transfer rate. Metal resistance genes were absent from most transconjugants, suggesting a minor role, if any, of metals in the spread of multidrug-resistant plasmids at the investigated sites.

Original languageEnglish
Article number997849
JournalFrontiers in Microbiology
Volume13
DOIs
StatePublished - 26 Oct 2022

Bibliographical note

Funding Information:
This study was supported by the Swedish International Development Cooperation Agency (Sida) to VI and ÅS, JG-O acknowledges the financial support from the International Science Program (ISP). C-FF acknowledges support from the Swedish Research Council Formas. DL would like to thank the EU and Swedish Research Council for funding in the frame of the collaborative international consortium (BIOCIDE) financed under the ERA-NET AquaticPollutants Joint Transnational Call (GA na869178). ÅS and EJ would like to thank the collaborative international consortium (PARRTAE) under the same call for funding. This ERA-NET is an integral part of the activities developed by the Water, Oceans, and AMR Joint Programming Initiatives. AF acknowledges financial support from the Centre for Antibiotic Resistance Research (CARe) at the University of Gothenburg and JPIAMR (2016-06503_3). Support was also obtained from the Swedish Research Council Dnrs 2014-02639, 2018-01874, and 2019-04202, and Swedish Research Links Dnr 2017-05423 to ÅS. Sequencing was performed with the support of the Centre for Translational Microbiome Research (CTMR), a collaboration between Karolinska Institutet, Science for Life Laboratory, and Ferring Pharmaceuticals.

Funding Information:
This study was supported by the Swedish International Development Cooperation Agency (Sida) to VI and ÅS, JG-O acknowledges the financial support from the International Science Program (ISP). C-FF acknowledges support from the Swedish Research Council Formas. DL would like to thank the EU and Swedish Research Council for funding in the frame of the collaborative international consortium (BIOCIDE) financed under the ERA-NET AquaticPollutants Joint Transnational Call (GA n869178). ÅS and EJ would like to thank the collaborative international consortium (PARRTAE) under the same call for funding. This ERA-NET is an integral part of the activities developed by the Water, Oceans, and AMR Joint Programming Initiatives. AF acknowledges financial support from the Centre for Antibiotic Resistance Research (CARe) at the University of Gothenburg and JPIAMR (2016-06503_3). Support was also obtained from the Swedish Research Council Dnrs 2014-02639, 2018-01874, and 2019-04202, and Swedish Research Links Dnr 2017-05423 to ÅS. Sequencing was performed with the support of the Centre for Translational Microbiome Research (CTMR), a collaboration between Karolinska Institutet, Science for Life Laboratory, and Ferring Pharmaceuticals. a

Publisher Copyright:
Copyright © 2022 Guzman-Otazo, Joffré, Agramont, Mamani, Jutkina, Boulund, Hu, Jumilla-Lorenz, Farewell, Larsson, Flach, Iñiguez and Sjöling.

Keywords

  • conjugative plasmid transfer
  • copper sulfate
  • Escherichia coli
  • horizontal gene transfer
  • IncN plasmid
  • multi-drug resistance
  • waterborne bacteria
  • zinc sulfate

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