Bovine herpesvirus 1 (BHV-1)is considered to be the most common viral pathogen found in bovine semen. Bovine herpesvirus 1 (BHV-1), the causative agent of infectious bovine rhinotracheitis (IBR), is considered to be the most common viral pathogen found in bovine semen. A total of 89 serum samples were screened for presence of BHV-1 antibodies and 26 sera were found to be positive by M-ELISA revealing an overall seroprevalence of 29.21%. For detection viral antigen in semen, a total of 101 samples were screened by applying direct FAT of which 33 (32.67%) were found to be positive for BHV-1 antigen. The viral genome in 101 semen samples was detected by employing PCR using two sets of primers from gB and gC genes coding for viral envelope. In gB gene based PCR, 47 (46.53%) semen samples were found to be positive. While in gC gene based PCR, 43 (42.57%) were found to be positive for presence of viral genome. In sequencing of gB-gene based PCR products a consensus sequence of 459 bp was obtained. The sequences of field isolate matches completely with the sequence of BHV-1.1. Finally, the study revealed presence of BHV-1 infection in breeding bulls of Gujarat.