This study was carried out to evaluate the effect of some disinfectants and antibacterial agents on Mycobacterium tuberculosis complex isolated from animal and human samples. The animal samples were collected from suspected animals from El-Sharkia, El- Menofya and El -Gharbiya governorate included lymph nodes (57 samples), raw milk samples (1250 sample) and human sputum samples (143) collected from the workers (attendance and cow-men) at infected farms. The bacteriological findings of the examined infected lymph nodes samples revealed that 31 sample out of 57 were harbored mycobacterium species with a percent of 54.4%, while 30 isolates of mycobacterium spp. were recovered on Lowenstein- Jenseen media with a percentage of 2.4% from the milk samples. On the other hand, the results of bacteriological examination of human sputum samples proved that 6 samples (4.2%) were harbored the acid fast bacilli by microscopical examination using Ziehl- Neelsen stain. The efficacy of both first and second line antituberculous drugs on the isolated strains were examined using method of proportion and showed that Rifampicin and Ethambutol had relative potent activity against the isolated mycobacteria also, Ciprofloxacin and Kanamycin had relative potent activity against the isolated mycobacteria. Evaluating the efficacy of disinfectants (phenol 5%-10% and ethyl alcohol 70%) using qualitative suspension test revealed that phenol 5% was effective at 20min exposure , phenol 10% was effective at 15min exposure and ethyl alcohol 70%  was effective at 20min.