A cross-sectional study was carried out to determine the seroprevalence and risk factors associated with Bovine viral diarrhea virus (BVDV) infection in non-vaccinated dairy and dual-purpose cattle herds from Ecuador. A total of 2,367 serum samples from 346 herds were collected from June 2008 through February 2009. A questionnaire, which included variables related to cattle, health, management measures, and the environment, was filled out in each herd. A commercial indirect enzyme-linked immunosorbent assay test was used to determine the seropositivity. A logistic regression model was used to determine risk factors at herd level. The individual seroprevalence for BVDV in non-vaccinated herds in Ecuador was 36.2% (857/2,367; CI(95%), 34.3-38.1%). The herd prevalence was 74% (256/346; CI(95%), 69.4-78.6%) and the intra-herd prevalence ranged between 11.1% and 100% (mean = 51.6%). The logistic regression model showed that the density of cattle farms in the area (more than 70%; OR, 1.94; CI(95%), 1.21-3.2) and the altitude (higher than 2,338 m above sea level; 2.33; CI(95%), 1.4-3.9) are potential risk factors associated with BVDV infection.