The aim of the present study was to determine the possible relationship between bovine viral diarrhoea (BVD) virus infection and the appearance of cervical mucous discharge (CMD) and the reproductive performance of cows in oestrus. For this purpose, CMD from 97 Holstein cows in oestrus was evaluated visually before artificial insemination (AI). Cows in oestrus were inseminated with frozen semen free from BVD virus (BVDV). Blood samples were tested by enzyme-linked immunoassay (ELISA) for antigen (Ag) and antibodies (Ab) of BVDV. The presence of the BVDV genome in cervical mucus samples was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). The presence of BVDV Ab, Ag or genome was not associated with abnormal cervical mucous discharge (A-CMD). However, the presence of BVDV Ag (but not of the BVDV Ab) in blood samples was associated with a lower first service conception rate (FSCR; 27.8 vs. 70.9%; P < 0.01), indicating that BVDV viraemia at the time of AI has a negative effect on the fertility of cows.