To the Editor: Since 1999, several zoonotic outbreaks of vaccinia virus (VACV) infection have been reported in cattle and humans in rural areas of Brazil. The infections have caused exanthematous lesions on cows and persons who milk them, and thus are detrimental to the milk industry and public health services (1,2). In Brazil during the last decade, VACV outbreaks have been detected from the north to the extreme south of the country (1–4). Because Brazil shares extensive boundaries with other South American countries, humans and cattle on dairy and beef-producing farms in those countries may be at risk of exposure to VACV. To determine if VACV has spread from Brazil to Argentina, we investigated the presence of VACV in serum samples from cattle in Argentina.