Abstract: Esherichia coli is one of the most important zoonotic enteric pathogen and most widely accepted indicator of fecal contamination in food and water. In present investigation, E. coli was isolated and identified in the faecal samples of pig, cattle and the farm workers handling the animals (20 each) using conventional and molecular methods and their virulence attributes and antibiogram were investigated. Total 40 isolates were tentatively identified as E. coli by conventional method while uspA gene was detected in only 17 (31%) isolates. A total of 46% isolates appeared pathogenic base on the virulence traits viz., haemolytic assay, congo red binding, haemagglutination, mannose resistant and mannose sensitive haemagglutination. In antibiogram study, all human and cattle isolates exhibited resistance to kanamycin, ampicillin, penicilline-G, cephalaxin, neomycin, streptomycin and ofloxacin while all cattle isolates were also found resistant to gentamycin and doxicycline. All the isolates from pigs exhibited resistance for gentamycin, kanamycin, penicillin-G, cephalexin and ampicilllin. Most important finding was the multiple drug resistance exhibited by majority of isolates involving as much as 5 antibiotics, which is an alarming situation posing threat to human and animal health.