Temperature and Nutrient Limitations Decrease Transfer of Conjugative IncP-1 Plasmid pKJK5 to Wild Escherichia coli Strains
Pallares-Vega, Rebeca; Macedo, Gonçalo; Brouwer, Michael S M; Hernandez Leal, Lucia; van der Maas, Peter; van Loosdrecht, Mark C M; Weissbrodt, David G; Heederik, Dick; Mevius, Dik; Schmitt, Heike
(2021) Frontiers in Microbiology, volume 12, pp. 1 - 13
(Article)
Abstract
Plasmid-mediated dissemination of antibiotic resistance among fecal Enterobacteriaceae in natural ecosystems may contribute to the persistence of antibiotic resistance genes in anthropogenically impacted environments. Plasmid transfer frequencies measured under laboratory conditions might lead to overestimation of plasmid transfer potential in natural ecosystems. This study assessed differences in the conjugative transfer
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of an IncP-1 (pKJK5) plasmid to three natural Escherichia coli strains carrying extended-spectrum beta-lactamases, by filter mating. Matings were performed under optimal laboratory conditions (rich LB medium and 37°C) and environmentally relevant temperatures (25, 15 and 9°C) or nutrient regimes mimicking environmental conditions and limitations (synthetic wastewater and soil extract). Under optimal nutrient conditions and temperature, two recipients yielded high transfer frequencies (5 × 10-1) while the conjugation frequency of the third strain was 1000-fold lower. Decreasing mating temperatures to psychrophilic ranges led to lower transfer frequencies, albeit all three strains conjugated under all the tested temperatures. Low nutritive media caused significant decreases in transconjugants (-3 logs for synthetic wastewater; -6 logs for soil extract), where only one of the strains was able to produce detectable transconjugants. Collectively, this study highlights that despite less-than-optimal conditions, fecal organisms may transfer plasmids in the environment, but the transfer of pKJK5 between microorganisms is limited mainly by low nutrient conditions.
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Keywords: AMR, antimicrobial resistance, environmental conditions, horizontal gene transfer, soil extract agar, synthetic wastewater, Microbiology, Microbiology (medical)
ISSN: 1664-302X
Publisher: Frontiers Media S.A.
Note: Funding Information: We gratefully acknowledge the support of the former students Xavier Gallego y van Seijen, Hana Pikov?, Gonzalo Monteoliva, Wesley Klaassen, Amaya G?mez, Dimitrios Maroulis, and Jenneke van der Draai for their relentless technical assistance in performing the conjugation assays. We extend our gratitude to professor S?ren J. S?rensen for kindly donating the donor strain. Funding. This work was performed in the cooperation framework of Wetsus, European Centre of excellence for sustainable water technology (www.wetsus.nl). Wetsus is co-funded by the Dutch Ministry of Economic Affairs and Ministry of Infrastructure and Environment, the European Union Regional Development Fund, the Province of Frysl?an and the Northern Netherlands Provinces. This work has also received funding from STOWA and from the European Union?s Horizon 2020 Research and Innovation Program under the Marie Sk?odowska-Curie [grant agreement no. 665874]. The authors like to thank the participants of the Wetsus research theme Source Separated Sanitation for the fruitful discussions and their financial support. Funding Information: This work was performed in the cooperation framework of Wetsus, European Centre of excellence for sustainable water technology (www.wetsus.nl). Wetsus is co-funded by the Dutch Ministry of Economic Affairs and Ministry of Infrastructure and Environment, the European Union Regional Development Fund, the Province of Fryslˆan and the Northern Netherlands Provinces. This work has also received funding from STOWA and from the European Union’s Horizon 2020 Research and Innovation Program under the Marie Skłodowska-Curie [grant agreement no. 665874]. The authors like to thank the participants of the Wetsus research theme Source Separated Sanitation for the fruitful discussions and their financial support. Publisher Copyright: © Copyright © 2021 Pallares-Vega, Macedo, Brouwer, Hernandez Leal, van der Maas, van Loosdrecht, Weissbrodt, Heederik, Mevius and Schmitt.
(Peer reviewed)