A sampling and metagenomic sequencing-based methodology for monitoring antimicrobial resistance in swine herds

Patrick Munk, Vibe Dalhoff Andersen, Leonardo de Knegt, Marie Stengaard Jensen, Berith Elkær Knudsen, Oksana Lukjancenko, Hanne Mordhorst, Julie Clasen, Yvonne Agersø, Anders Folkesson, Sünje Johanna Pamp, Håkan Vigre, Frank Møller Aarestrup

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Abstract

Objectives
Reliable methods for monitoring antimicrobial resistance (AMR) in livestock and other reservoirs are essential to understand the trends, transmission and importance of agricultural resistance. Quantification of AMR is mostly done using culture-based techniques, but metagenomic read mapping shows promise for quantitative resistance monitoring.
Methods
We evaluated the ability of: (i) MIC determination for Escherichia coli; (ii) cfu counting of E. coli; (iii) cfu counting of aerobic bacteria; and (iv) metagenomic shotgun sequencing to predict expected tetracycline resistance based on known antimicrobial consumption in 10 Danish integrated slaughter pig herds. In addition, we evaluated whether fresh or manure floor samples constitute suitable proxies for intestinal sampling, using cfu counting, qPCR and metagenomic shotgun sequencing.
Results
Metagenomic read-mapping outperformed cultivation-based techniques in terms of predicting expected tetracycline resistance based on antimicrobial consumption. Our metagenomic approach had sufficient resolution to detect antimicrobial-induced changes to individual resistance gene abundances. Pen floor manure samples were found to represent rectal samples well when analysed using metagenomics, as they contain the same DNA with the exception of a few contaminating taxa that proliferate in the extraintestinal environment.
Conclusions
We present a workflow, from sampling to interpretation, showing how resistance monitoring can be carried out in swine herds using a metagenomic approach. We propose metagenomic sequencing should be part of routine livestock resistance monitoring programmes and potentially of integrated One Health monitoring in all reservoirs.
Original languageEnglish
Article numberdkw415
JournalJournal of Antimicrobial Chemotherapy
Volume72
Pages (from-to)385-392
Number of pages8
ISSN0305-7453
DOIs
Publication statusPublished - 2016

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