In Korea livestock farms breeding cattle have been suffering from re-emerging problems of Brucella (B.) abortus infection while steady decline of bovine brucellosis. Therefore, this study underscored the identification and association of etiological agent of brucellosis in cattle in South Korea. The incidence of brucellosis in cattle was analyzed by bacteriological and molecular methods in 187 brucellosis-suspicious farms of 11 regions between 2018 and 2020. Brucella isolates from various specimens were identified by Brucella-specific multiplex PCR. Epidemiological data were collected by local official veterinarians through history taking from farmers and animal data systems. In 230 of 560 cattle (40.9%) and 94 of 187 farms (50.3%), a total of 313 B. abortus were isolated from various specimens, the majority of isolates were from supramammary lymph node (41%). In epidemiological findings, the majority of positive cases were mainly caused by resurgence (43.7%) and unknown (37.2%). Of 94 positive cases isolated B. abortus, abortion in cattle infected by B. abortus occurred in 51 farms (54.3%) where led to resurgence in 30 farms and environmental survival of B. abortus in 9 farms. Consequently, these findings revealed the existence of etiological agent of bovine brucellosis in Korea, which still occurred at low levels in distinct regions where are allowed to call for persistent biosecurity. Thus, we highlight that brucellosis in Korea needs to take more effective control strategies with potential evidence. Moreover it is ultimately important to maintain a constant monitoring for eradication of brucellosis.