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In this work, Mycobacterium bovis isolated and identified by microbiological culture and RT-PCR from emergency slaughtered aburden Angus cattle suffering from chronic mastitis and respiratory manifestations in private farm not perform tuberculin test in New Valley governorate, Egypt. Furthermore, other contact cattle, sheep and human in the farm examined by ELISA, found that 22.2% (4/18) cattle in the farm were positive, with complete absence of infection in the sheep and contact persons in the farm. The high percent of M. bovis was detected by RT-PCR (83.33%) followed by microbiological culture (72.22%) in slaughtered cattle. The highest percent of tubercles lesion observed in thoracic cavity followed by the lung then the udder. From our study we can concluded that, M. bovis isolated and identified from imported Aburden Angus cow in New Valley Governorate. Also, ELISA test need further evaluation before using as an exploratory test for diagnosis of BTB in animal and human.