Aim:Colibacillosis is considered as one of the major bacterial infections in avian pathology. The excessive use of antibiotics reduced their effectiveness, which eventually led to the risk of emergence of antibiotic resistance. The aim of this study was to isolate, identify and serotype the pathogenic Escherichia coli strains and to determine their antibiotic susceptibility.Materials and Methods:A total of 180 samples from different organs of broilers with colibacillosis lesions were collected (liver, spleen, lung, and heart) in center of Algeria. The isolation and identification of E. coli were carried out using conventional techniques. Then, these strains were serotyped and tested over 13 antibiotics.Results:A total of 156 strains of E. coli were isolated. Serotyping results showed that 50 strains belong to 3 serotypes (23 for O1, 11 for O2, 16 for O78) which represent 32% of isolates. The antimicrobial susceptibility test, presented high level of resistance to tetracyclines (94.12%), flumequine (91.5%), sulfamethoxazole-trimethoprim (88.89%), enrofloxacin (86.27%), nalidixic acid (85.62%), ampicillin (83.01%) and doxycycline (75.81%), medium level resistance to chloramphenicol (39.22%), and amoxicillin-clavulanic acid (43.13%). All the strains were susceptible to cefotaxime, excepting three, which presented an extended spectrum β-lactamase (ESBL). In addition, the results of multi-resistance showed that all the strains were resistant at the minimum to two antibiotics and 66.66% of strains were resistant to at least seven antibiotics.Conclusion:The antibiotic resistance continues to rise at an alarming rate, and the emergence of ESBL is considered as a threat for public health.