The Frutillar 82, F nasal 90, F vaginal 90, Puente Alto 77, VMLB 79, and Bio Bio 92 isolates of bovine herpesvirus 1 (BHV-1) from Chile were analysed by Eco RI, Bam HI and Hind III restriction enzymes (RE). The Los Angeles (LA) strain was used as reference. The results indicated that the fingerprints of F nasal 90 and F vaginal 90 were identical to the three RE used, but they were different from the other isolates. Major differences were found for Puente Alto 77 and VMLB 79 isolates with Eco RI, and for Puente Alto 77 with Hind III compared with the other Chilean isolates. It is concluded that Chilean BHV-1 isolates obtained from diverse lesions and geographic locations are genomically different from each other and from the other strains described in the literature.