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Identification of diverse antibiotic resistant bacteria in agricultural soil with H218O stable isotope probing and metagenomics

Marcela Hernández, Shamik Roy, C. William Keevil, Marc G. Dumont
doi: https://doi.org/10.1101/2023.01.24.525391
Marcela Hernández
aSchool of Biological Sciences, University of Southampton, Southampton SO17 1BJ, UK
bSchool of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK
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Shamik Roy
bSchool of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK
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C. William Keevil
aSchool of Biological Sciences, University of Southampton, Southampton SO17 1BJ, UK
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Marc G. Dumont
aSchool of Biological Sciences, University of Southampton, Southampton SO17 1BJ, UK
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  • For correspondence: m.g.dumont@soton.ac.uk
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Abstract

Background In this study, we aimed to identify bacteria able to grow in the presence of several antibiotics including the ultra-broad-spectrum antibiotic meropenem in a British agricultural soil, by combining DNA stable isotope probing (SIP) with high throughput sequencing. Soil was incubated with cefotaxime, meropenem, ciprofloxacin and trimethoprim in 18O-water. Metagenomes and the V4 region of the 16S rRNA gene from the labelled “heavy” and the unlabelled “light” SIP fractions were sequenced.

Results After incubations, an increase of the 16S rRNA copy numbers in the “heavy” fractions of the treatments with 18O-water compared with their controls was detected. The treatments resulted in differences in the community composition of bacteria. Members of the phyla Acidobacteriota (formally Acidobacteria) are highly abundant after two days of incubation with antibiotics. Several Pseudomonadota (formally Proteobacteria) including Stenotrophomonas were prominent after four days of incubation. Furthermore, a metagenome-assembled genome (MAG-1) from the genus Stenotrophomonas (90.7% complete) was retrieved from the heavy fraction. Finally, 11 antimicrobial resistance genes (ARGs) were identified in the unbinned-assembled heavy fractions, and 10 ARGs were identified in MAG-1. On the other hand, only two ARGs from the unbinned-assembled light fractions were identified.

Conclusions The results indicate that both non-pathogenic soil-dwelling bacteria as well as potential clinical pathogens are present in this agricultural soil, and several ARGs were identified from the labelled communities, but it is still unclear if horizontal gene transfer between these groups can occur.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 24, 2023.
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Identification of diverse antibiotic resistant bacteria in agricultural soil with H218O stable isotope probing and metagenomics
Marcela Hernández, Shamik Roy, C. William Keevil, Marc G. Dumont
bioRxiv 2023.01.24.525391; doi: https://doi.org/10.1101/2023.01.24.525391
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Identification of diverse antibiotic resistant bacteria in agricultural soil with H218O stable isotope probing and metagenomics
Marcela Hernández, Shamik Roy, C. William Keevil, Marc G. Dumont
bioRxiv 2023.01.24.525391; doi: https://doi.org/10.1101/2023.01.24.525391

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