Nosocomial infections are frequent complications of hospitalization, caused by opportunistic pathogens that gain access to hosts undergoing invasive procedures. This study was carried out to investigate the presence of antibiotic resistance bacteria from hospital environment of Damietta, Egypt. Two hundred and six clinical bacterial isolates were collected from different samples (sink, floor, bed, bed cover, toilet floor, bed pan, ward wall and hospital staff apparel (protective gowns, hand groves and face-shield) within reception hall, maternity ward, convalescing wards, surgical theaters, intensive care, dental unit, pharmacy and laboratory) from (Kafr Saad General Hospital and El-Azhar University Teaching Hospital). Four bacterial species were identified as Bacillus subtilis, Bacillus cereus, Klebsiella pneumoniae and Enterobacter aerogenes using standard morphological, biochemical tests and sequencing of 16S rRNA gene. Eleven antibiotics (Trimethoprim/sulphamethoxazole (25 μg); Rifampicin (5 μg); Piperacillin/tazobactam (110 μg); Ofloxacin (5 μg); Nitrofurantoin (300 μg); Imipenem (10 μg); Gentamicin (120 μg); Ceftriaxone (30 μg); Cefotaxime (30 μg); Amikacin (30 μg) and Amoxicillin/clavulanic acid (30 μg) were tested against the bacterial isolates using disc diffusion method to determine the multi-drug resistance bacteria. Imipanem was found to be as the most effective drug against Klebsiella pneumoniae while ofloxacin, amikacin, piperacillin/tezobactam, gentamicin and rifampcin had low effect against Klebsiella pneumoniae and no effect of amoxicillin/clavulanic acid, cefotaxime, ceftriaxone, nitrufurantion and trimethoprim/sulehamethoxazole was recorded against Bacillus subtilis.