Escherichia coli (E. coli) causes various infections in humans and animals. Animals and humans can get a variety of diseases from E. coli. Because of its tendency to build biofilms, Escherichia coli is more resistant to antibiotics and can cause chronic and recurrent infections. In connection to antibiotic resistance, this study assessed the biofilm-forming capacity of E. coli isolated from extra intestinal infections in humans and chickens. The numbers of total samples that isolated from humans 8(26.67%) and chickens 12(40%) were twenty and harvested to determine of the biofilm forming by the test tubes, congo red agar method, appear all E. coli isolated were positive to biofilm. The pattern of resistance of these strains mentioned that E. coli from chicken samples was resistant to Nalidixic acid (100%), Ciprofloxacin (58.4%), Amikacin (58.4%), Nitrofurantoin (58.4%) Cefixime (16.6%), Ceftazidime (25%), Tetracycline (25%) and Gentamicin (33.3%). E. coli from human clinical samples was resistant Amikacin100%, Nitrofurantoin100%, Nalidixic acid, Ciprofloxacin 62.5%, Tetracycline 62.5%, Cefixime 50%, Gentamicin 25% and Ceftazidime 12.5%. Additionally, both human and chiken samples frequenently contained multiresistanant E.coli isolates. According to the study's findings, hens may serve as reservoirs for the spread of microorganisms resistant to antibiotics to people. Moreover, any ability of E. coli to build biofilms can raise the isolates' profile of antibiotic resistance.