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From the world ' s highlight on dangers of tuberculosis as a zoonotic and debilitating disease threatening animal and human health beside economy, this research aimed to determine the prevalence of Mycobacterium bovis from buffaloes' mastitic milk by isolation on Lowenstein-Jensen medium (LJ medium), Ziehl-Neelsen staining; identification by different biochemical tests and polymerase chain reaction (PCR) at 500bp diagnostic for M. bovis, as well as (2) to determine the therapeutic efficacy of the various antimicrobials (9 types) on the isolated M. bovis by using different antimicrobial sensitivity plate method. A total number of 100 samples of mastitic milk from native lactating buffaloes' breed were collected aseptically from 4 private farms in Cairo-Alexandria desert road in the north part of the Egypt and subjected to bacteriological and molecular examination for M. bovis followed by their antibiogram profile. Seven (7%) isolates were identified as M. bovis from totally examined 100 milk samples and isolated M. bovis showed resistance to ciprofloxacin; gentamycin and sulphamethoxazole-trimethoprim while were intermediate sensitive to both of erythromycin and norfloxacin but on the contrary were sensitive to amikacin; cefotaxime; clindamycin and streptomycin. It can be concluded that M. bovis can be the causative agent of mastitis in Egyptian Buffaloes and could be a potential risk for zoonotic transmission to man as well as economic losses. So, strict hygienic regulations and novel diagnostic tools should be used for prevention and detection of tuberculous mastitis in buffaloes' farms. The general public health should also be intensely warned from consuming raw or unpasteurized milk. All these prophylactic measures will eventually lead to a positive impact on public health.

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