From January 1990 through March 1991, 189 armadillos (Dasypus novemcinctus) were collected from Brevard and Glades Counties in southern Florida (USA). The sera were analyzed for hemagglutination-inhibition (HAI) antibodies against St. Louis encephalitis (SLE) and eastern equine encephalomyelitis (EEE) viruses. None of the armadillos had detectable HAI antibody to EEE virus, but 59 (31%) had antibodies against SLE virus. Sera from 31 of the HAI-positive armadillos contained significant levels of neutralizing (NT) antibody to SLE virus. Armadillos captured during the 1990 SLE human epidemic in south Florida had a greater prevalence of HAI and NT antibody to SLE virus than did animals captured before the start of the epidemic. This is evidence that armadillos were fed on by mosquitoes infected with SLE virus. We propose that armadillos may be involved in the SLE amplification and transmission cycles in Florida.