A herpesvirus was isolated from Thomson's gazelle (Gazella thomsoni) kept at a zoological garden in Japan during an outbreak of epizootic acute encephalitis. The virus, gazelle herpesvirus 1 (GHV-1), was serologically related to equine herpesvirus 1 (EHV-1). However, DNA fingerprints of GHV-1 were different from those of EHV-1 and other equine herpesviruses. Southern hybridization with probes of cloned BamHI fragments derived from UL and US segments of EHV-1 revealed differences in the DNA restriction profiles throughout the entire genome. Nucleotide sequences were determined for a conserved region of an essential envelope glycoprotein B (gB) gene and a type-specific glycoprotein G (gG) homologue gene. The predicted amino acid sequence of GHV-1 gB showed 97, 92, 61, and 57% identity to EHV-1, EHV-4, feline herpesvirus, and pseudorabies virus, respectively, indicating that GHV-1 was closer to EHV-1 than any other herpesvirus. The GHV-1 gG gene showed 93.2, 92.3, and 53% identity to EHV-1, EHV-8, and EHV-4 gGs, respectively. GHV-1 was virulent to suckling mice of the ICR strain by intracerebral inoculation and was virulent to 4-week-old BALB/c mice by intranasal inoculation, causing neurological symptoms and death. We conclude that GHV-1 is a new type of equine herpesvirus with strong neurotropism.