A study was conducted on ‘Escherichia coli’ to reveal its presence in poultry and poultry environment in Jammu (India). A total of 200 samples (90 of poultry droppings, 45 waterers, 40 feeders and 25 poultry handlers) were processed, of which 148 samples yielded E. coli isolates. The isolation rate of E. coli from poultry droppings was 92.22% while from poultry environment was 59%. The 148 isolates of E. coli were further characterized by performing multiplex polymerase chain reaction for the genes, stx1, stx2, eaeA and ehxA. 16 of 148 (10.81%) of isolates were carrying the virulence genes. Out of them 13 isolates were from poultry and three isolates were obtained from waterers. Thirteen isolates from poultry droppings were positive for virulence genes, eight (61.5%) harbored eaeA gene, while four harbored stx2 and eaeA, both. One of the isolate demonstrated the presence of stx2 gene, alone. A total of three out of 36 isolates from waterers revealed the presence of eaeA gene. All the isolates were negative for the ehxA gene. Also, none of the isolate from poultry feed or poultry handlers carried virulence genes. The antibiotic sensitivity assay of isolates was done against 11 antibiotics commonly practiced in poultry medicine revealed maximum resistance to Enrofloxacin (100%), followed by Cefotaxime (91.89%), while maximum sensitivity was noted to Amoxicillin/Clavulanic acid (95.27%).