ABSTRACT. A nationwide survey of bovine leukemia virus (BLV) infection was conducted among dairy and beef breeding cattle in Japan from 2009-2011 using an enzyme-linked immunosorbent assay. Of a total of 20,835 cattle tested, 35.2% were seropositive for BLV and the animal type-level seroprevalences in dairy and beef breeding cattle were 40.9 and 28.7%, respectively. By the time animals were 1 year old, 21.0% of dairy and 13.7% of beef breeding cattle were considered infected. Our findings indicate that BLV is widespread among dairy and beef breeding cattle in Japan with the BLV seroprevalences approximately 10-and 4-fold higher, respectively, than previously reported for 1980-1982 in Japan. KEY WORDS: bovine leukemia virus, cattle, nationwide survey, seroprevalence.