A cross-sectional study was conducted on 1138 heads of cattle at Kombolcha ELFORA meatrnprocessing plant, South-Wello Administrative Zone, Amarha Regional State from Novemberrn2007 to May 2008. The objectives of the study were to estimate the prevalence of bovinerntuberculosis in cattle slaughtered at Kombolcha ELFORA meat processing plant, and tornisolate and characterize the strains of mycobacteria from tuberculousis suspicious lesions.rnDetailed post mortem examination, bacteriological culturing, regions of difference-basesrnpolymerase chain reaction and spoligotyping were used. The prevalence of bovinerntuberculosis was 5.0 % (57/1138) in cattle slaughtered at Kombolcha ELFORA meatrnprocessing plant on the basis of detailed post-mortem examination. There was no significancerndifference in prevalence between male and female (P > 0.05), as well as among the agerngroups (P > 0.05). Mycobacteria were able to be isolated from 28 of the 57 cattle with grossrnlesion. A total of 49 isolates were recovered from the different tissues of these 28 cattle, andrn20 isolates showed signal for M. tuberculosis complex species of which 19 were M. bovisrnwhile one isolate was M. tuberculosis. Further characterization of the strains of M. bovisrnusing spoligotyping revealed the presence of five different cluster of spoligotyping pattern,rnwhich include Ethiopian M. bovis train 1, SB1176, SB0134, SB0133 and new strain. Thernbinary number representation of the new strain isrn1100000101010110111111111111111101111100000 where 1 indicates the presence of arnspacer and 0 indicates a loss. The new spoligotyping was not reported previously fromrnelsewhere to the M. bovis database (www.mbovis.org). The findings of this study indicatedrnthat tuberculous lesions were common in cattle slaughtered at Kombolcha ELFORA meatrnprocessing plant, and hence detailed post mortem meat inspection procedures are required tornminimize the risk of its transmission to the public through meat consumption. In addition,rnappropriate cooking of meat is required before consumption.