New technologies in the field of vaccinology arise as necessity for treatment and control of many diseases. Currently modified live virus and inactivated vaccines used for Bovine Herpesvirus-1 (BoHV-1) have several disadvantages. Previous works for preventive treatment of BoHV-1 with DNA based vaccines have demonstrated the capability to induce humoral and cellular immune response. Nevertheless, it is well known that “naked” DNA induces low immunogenic response. Thus, loading of antigen encoding DNA sequences in liposomal formulations targeting dendritic cell receptors could be a promising strategy to better activate these antigen presenting cells (APC). In this work, DNA based vaccine encoding the truncated version of gD glycoprotein (pCIgD) of BoHV-1 was investigated alone and upon encapsulation on liposomal formulation coated with MANα1-2MAN-PEG-DOPE and LPS from Brucella ovis (pCIgD-Man-L) in mice and cattle assay. Results showed that the use of pCIgD-Man-L was capable to enhance the immune response in both animal models. Significant levels of humoral immunity were achieved when total antibody titers and isotypes were detected in sera and mucosa. For cellular immunity, specific viral lymphoproliferation was detected in the animals inoculated with pCIgD-Man-L. In addition, positively modulation of CD40 molecules on the surface of bovine dendritic cells (DCs) was observed when cells were stimulated and activated with vaccine formulations. When challenge assay was performed, bovines inoculated with pCIgD and liposome decorated with MANα1-2MAN-PEG-DOPE elicited better protection and diminished viral excretion. The results demonstrate the targeting of the MANα1-2MAN coated liposomes toward dendritic cells and their ability to boost the immunogenicity according to an adjuvant effect that results in long-lasting immunity. Liposome decorated with MANα1-2MAN-PEG-DOPE were for the first time tested as DNA based vaccine in cattle as preventive treatment of BoHV-1. These results open up new perspectives for the design of vaccine for the control of bovine rhinotracheitis.