Members of the genus Brucella are facultative intracellular bacteria and cause brucellosis, a chronic disease in humans and abortion in animals. In this study, we tested sera for brucellosis of 15 Hanwoo farms in the western part of Gyeong-buk province, resulting 5 farms were brucellosis positive in 2008. We collected blood from 277 heads in the brucellosis positive 5 farms, and performed serological diagnosis, brucella positive cattle which had shown higher than 200 antibody titer in tube agglutination test were slaughtered, supramammary lymph nodes were collected, and Brucella spp. wild type isolation and identification were performed. From these results, 15 of Brucella spp. wild type strains were isolated and all strains were identified as B. abortus biotype 1 by biological and molecular analysis. In the antimicrobial susceptibility test, all 15 strains had a similar susceptibility and resistance pattern. This study may be useful for bacteriological and epidemiological understanding of cattle brucellosis in Korea.