Antimicrobial resistance (AMR) is a global health crisis, with Enterococcus faecium ranked by WHO as a high-priority pathogen due to limited treatment options. DYNAMIC-E aims to map the prevalence, resistance profiles, and gene transfer potential of multidrug-resistant Enterococcus spp. across cattle from different production systems, farmers, veterinarians, and farm environment in Northern Portugal, while assessing the influence of seasonality and temperature on AMR dynamics. Preliminary results from 50 cattle samples revealed unexpectedly high resistance, including to the last-resort antibiotic linezolid. The strategy involves characterizing the most relevant species (E. faecalis, E. faecium) from different breeds, farms and seasons; and evaluating conjugative transfer of key resistance genes under different temperatures. The project will assess the impact of temperature on horizontal gene transfer and AMR dynamics. Findings will support One Health surveillance and inform intervention strategies in order to safeguard antibiotics as linezolid to treat severe human infections. The project will also foster capacity building by involving one PhD student and one undergraduate student.