Neotropical bats were collected from different life zones in Guatemala in 1983 and 1984 to determine the presence and distribution of antibody against 10 viruses. Bats were collected with mist nets at 13 sites in eight departments and 332 serum specimens were obtained for testing for neutralizing (N) antibody by the plaque-reduction neutralization test. Eighty-seven (26%) of the 332 bats from 16 (38%) of 42 bat species sampled were serologically positive for five of six arboviruses and for two other viruses tested. Antibodies against Venezuelan equine encephalitis (VEE) variant I-A/B, eastern equine encephalitis, western equine encephalitis, St. Louis encephalitis, vesicular stomatitis, Tacaribe, and Rio Bravo viruses were detected in resident species of bats. However, N antibodies against the enzootic strain of VEE (Mena II, variant I-E) or Nepuyo viruses were not detected.