On-farm biosecurity measures are an important part of a control plan to minimize the introduction and spread of infectious diseases, such as equine influenza, in an equine facility. It can be challenging, however, to evaluate the efficacy of biosecurity measures under field conditions. We used an agent-based computer simulation model to describe the impact of: i) preventive vaccination; ii) reduced horse-to-horse contact; and iii) a combination of vaccination and reduced contact during an outbreak of equine influenza in a simulated horse facility. The model demonstrated that the most effective intervention was a combination of a high proportion of recently vaccinated horses and a substantial reduction in horse-to-horse contact once equine influenza had been identified in the facility. This study highlights the importance of compliance when implementing biosecurity measures, such as facility-level infection control practices, on horse farms.