Antimicrobial resistance (AMR) has evolved in environmental bacteria over billions of years, producing a vast reservoir of AMR genes that can potentially be transferred to clinical pathogens. Agricultural soils are exposed to antibiotics through the use of manures or sewage sludge as fertiliser, or irrigation with reclaimed water. There is a concern that such exposure may promote AMR in food producing environments, increasing the likelihood of AMR transmission to the human microbiome via contaminated crops or environmental matrices. There is a growing body of research using in vitro models to establish the minimal selective concentrations (MSCs) of antibiotics in the environment, but there is very little evidence relating to selection by antibiotic residues jn situ in soils. The field scale trials undertaken at Agriculture and Agri-Food Canada (AAFC) over the last 10-20 years offers a unique opportunity to investigate the effects of antibiotic application on soil communities and AMR, and how soils recover from long-term application of antibiotics. In this project, state of the art metagenomic and bioinformatic approaches will be applied to determine the evolutionary effects of antibiotic residues on the soil resistome. This will be investigated at three timepoints: short-term exposure (7 & 30 days after application); long-term exposure (after 10 years of annual exposure); and after cessation of antibiotic application (1, 2, and 3 years post-exposure). Macrolide and fluoroquinolone antibiotics will be studied, which have very different environmental fates and which have both been flagged as priority substances of concern by the EU Water Framework Directive's Hazardous Compound Watch List. Furthermore, we will assess whether or not in vitro experiments are predictive of selection in soils in situ. This work will provide the most comprehensive assessment of AMR evolution in antibiotic amended soils to date.