This survey was carried out to determine the seroprevalence of nasal infestation by sheep bot fly, Oestrus ovis L., and to identify the risk factors associated with the disease in flocks in southwestern Spain. In total, 5,878 sera samples of adult sheep were collected at random in 551 farms from four provinces in the southwestern Spain: Badajoz, Cáceres, Córdoba, and Sevilla. Sera were tested by enzyme-linked immunosorbent assay for O. ovis antibodies, by using a crude L2 larval as antigen. The seropositive mean prevalence was 69.30%, and mean percentage of optical densities was 61.83%. There were significant differences between the provinces studied; Córdoba and Sevilla were the provinces with more infested animals and higher seroprevalences. The correlation between seroprevalence and percentage of antibodies by farms was significant. There were only 18 farms free of seropositive animals, and 115 of the total 551 farms had all sampled animals seropositive, an indication of the high importance of this parasitosis in the investigated areas. Altitude, latitude, flock size, and ovine population density were the potential risk factors associated with the detection of O. ovis antibodies. Those animals breeding in regions located at low altitudes (<500 m), meridian latitudes (<39.5° N), and on farms with medium-to-large flock size (>250 sheep) and high ovine population density (>100 sheep per km²) were more likely to be seropositive. These findings confirm that these studied factors should be considered as potential risk factors to the presence of O. ovis in ovines from southwestern Spain.