Journal
ENVIRONMENTAL SCIENCE & TECHNOLOGY
Volume 56, Issue 16, Pages 11398-11408Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.est.2c02686
Keywords
lateral gene transfer; antibiotic resistance gene; soil microbiome; mating; cattle manure
Categories
Funding
- Dutch Ministry of Economic Affairs and Ministry of Infra [665874]
- European Centre of excellence for sustainable water technology
- Dutch Ministry of Economic Affairs and Ministry of Infrastructure and Environment
- European Union Regional Development Fund
- Province of Fryslan
- Northern Netherlands Provinces
- European Union [665874]
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This study aimed to quantify and identify new plasmid-acquiring bacteria resulting from manure application to soils to assess the risk of horizontal gene transfer in the environment. The results showed that gene transfer from fecal to soil bacteria occurred despite suboptimal conditions, but these events were rare and mainly happened shortly after manure application, with the plasmid not colonizing the soil community.
The quantification and identification of new plasmid-acquiring bacteria in representative mating conditions is critical to characterize the risk of horizontal gene transfer in the environment. This study aimed to quantify conjugation events resulting from manure application to soils and identify the transconjugants resulting from these events. Conjugation was quantified at multiple time points by plating and flow cytometry, and the transconjugants were recovered by fluorescence-activated cell sorting and identified by 16S rRNA sequencing. Overall, transconjugants were only observed within the first 4 days after manure application and at values close to the detection limits of this experimental system (1.00-2.49 log CFU/g of manured soil, ranging between 10(-5) and 10(-4) transconjugants-to-donor ratios). In the pool of recovered transconjugants, we found amplicon sequence variants (ASVs) of genera whose origin was traced to soils (Bacillus and Nocardioides) and manure (Comamonas and Rahnella). This work showed that gene transfer from fecal to soil bacteria occurred despite the less-than-optimal conditions faced by manure bacteria when transferred to soils, but these events were rare, mainly happened shortly after manure application, and the plasmid did not colonize the soil community. This study provides important information to determine the risks of AMR spread via manure application.
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