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Abstract Bacterial resistance to penicillin and cephalosporins is an increasing problem in human medicine and an emerging problem in veterinary medicine, particularly in certain Gram-negative organisms. Bacteria have become resistant to antimicrobial agents as a result of chromosomal changes or the exchange of genetic material via plasmid and so, treatment depends on the disease and should be guided by laboratory analysis of the antibiotics sensitivities of the infecting strain of E. coli. The emergence of multidrug resistant bacteria is mainly associated with β- lactamase production. The current research aims to investigate the occurrence of antibiotic resistance genes, TEM, SHV, OXA and AmpC β-lactamases , in E.coli strains from human and poultry sources. Sixty E. coli strains were studied, thirty five of human origin (recovered from hospitalized patients with gastrointestinal tract disturbances) and twenty five of animal origin (obtained from trachea of poultry with respiratory manifestations). The strains were inoculated onto MacConkey’s agar medium and incubated at 37°C for 24hrs. Lactose fermenter colonies were subcultured onto eosin methylene blue agar medium and incubated at 37°C for 24hrs. Developed colonies were subjected to indole, methyl red, Voges |