The results of molecular-genetic analyses of milk sample are presented in this article. The experimental studies were carried out in the dairy cattle of livestock farms of the Vologda region suspected to the respiratory infections. Samples of milk were obtained from the cows with the following clinical signs: cough and mucosal discharges from the nasal cavity. The laboratory studies were conducted in the virology laboratory of Federal scientific center-all-Russian research Institute of experimental veterinary medicine. Polymerase chain reaction and Reverse transcription polymerase chain reaction were used for the detection of the genome of the viruses. The samples were tested for the presence of following viral infections: bovine herpesvirus, bovine viral diarrhea virus - mucosal disease, parainfluenza-3 virus and coronavirus. A comparative analysis of the nucleotide sequences of the studied virus and the reference strains of the GenBank database showed the similarity of the detected virus in cow milk of the Vologda region with the Bovine herpesvirus-5 (BoHV-5). During the differential diagnostics, the coronavirus, bovine viral diarrhea virus and parainfluenza-3 virus were excluded. Thus, as a result of the studies, Bovine herpesvirus-5 (BoHV-5) was detected for the first time in the milk of cows of livestock farms of the Vologda region.