The area affected by the disease caused by Madariaga virus has been expanding recently. Given the geographic location of the endemic regions that overlap with tourist destinations, as well as the confirmed possibility of importing the disease, we cannot rule out the risk of appearance of this disease in Russia. The aim of the study was to analyse the properties of Madariaga virus (Alphavirus genus, eastern equine encephalomyelitis virus complex) and some epidemiological and epizootological characteristics of the virus-associated disease which manifests itself in diffuse inflammation of brain and spinal cord. Based on both ecological and molecular genetic characteristics of Madariaga virus, it is classified as an individual virus which is part of the eastern equine encephalomyelitis virus complex. There is evidence that this pathogen can cause epizootic outbreaks in horses, infect other types of mammals (rats, bats), as well as, potentially, birds and reptiles. The reservoir of infection is the short-tailed cane mouse (Zygodontomys brevicauda). Human cases have also been described, and the potential route of infection was identified as transmission via mosquito bites. Vectors of pathogen transmission are mosquitoes of the Culex, Aedes, and Psorophora genera. Serological studies carried out in Panama revealed the presence of antibodies to Madariaga virus in 2–5% of the examined patients, which indicates the existence of asymptomatic infection along with clinically apparent infection. Phylogenetic analysis of strains isolated from infected people showed that the strains belong to lineage III of the eastern equine encephalomyelitis virus subtype common in Central and South America. The paper summarises the results of assessment of potential risk factors of Madariaga virus infection in endemic regions that was performed using the Akaike information criterion. Risk groups in endemic regions are represented by farm workers and fishermen. The results of the studies show that the area affected by the disease is expanding, and the strains that belong to genetic lineage III of the eastern equine encephalomyelitis virus pose the greatest epidemic risk.