Replication-defective human adenoviruses type 5 (HAd5) expressing the bovine herpesvirus type 1 (BHV-1) glycoprotein gC or gD under the control of the human cytomegalovirus immediate-early promoter/enhancer (AdCMVgC or AdCMVgD) or the 5' regulatory region of the human desmin gene (AdDESMgC or AdDESMgD) were generated. A preliminary experiment performed on rabbits showed that the intranasal administration of AdCMV elicited higher levels of BHV-1 neutralizing antibodies than the intramuscular administration of AdDESM. The obtained results allowed to select the replication-defective AdCMVgC and AdCMVgD for further assessment of their potential as a recombinant vaccine in cattle. Calves were injected intranasally twice 3 weeks apart with either AdCMVgC or AdCMVgD or a combination of these two recombinants or a commercially available live vaccine for comparison. The highest BHV-1 neutralizing antibody titres were obtained with AdCMVgD followed by the live vaccine and to a lower extent with the combination of the two recombinants (AdCMVgC+AdCMVgD). Calves were protected against intranasal BHV-1 challenge performed 3 weeks after the second immunization. In view of the obtained results, recombinant HAd5 may be developed as an intranasal vaccine vector in cattle administrated either alone or sequentially with non-human adenovirus-based vectors.