We investigated the presence of Mycobacterium spp. in livestock in northern China. Of the 163 clinical samples selected for this study, 20 were from throat swabs of dairy cows, and 143 were tissue samples (including lung tissue from one reindeer, hilar lymph node tissue from 55 cows, and liver tissue from 87 sheep). A total of 41 mycobacterial isolates were identified including two isolates of M. caprae and 39 non-tuberculous mycobacteria (NTM) isolates. Multi-locus variable-number tandem repeat analysis (MLVA) profiles of the two M. caprae isolates proved to be unique. This is the first report of M. caprae isolates from livestock in China. This study also confirms previous reports that NTM is common in livestock in northern China.