In 1983, 17 virus strains were isolated from mosquitoes collected during an outbreak of western equine encephalitis in Santa Fe Province, Argentina. Strains of western equine encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, and Antequera viruses were isolated, as were several bunyaviruses of the California and Bunyamwera serogroups and a new vesiculovirus. Complement fixation and neutralization tests were used to identify the California serogroup virus as a subtype of Melao virus, the Bunyamwera serogroup virus as a subtype of both Maguari and Playas viruses, and the vesiculovirus as a newly recognized agent for which the name Calchaqui virus is proposed. A limited serosurvey of horses and humans in Santa Fe Province and horses from the adjacent Santiago del Estero Province was performed to determine the prevalence of neutralizing antibody to the subtypes of Melao and Maguari viruses and to Calchaqui virus. The high prevalence of antibodies to these three agents indicates the need for further studies of their disease potential in horses, because they are closely related to several other viruses that are known equine pathogens.