The aim of the study was to carry out the isolation, prevalence and to determine the in-vitro pathogenicity and antibiogram of Escherichia coli. A total of 170 samples comprising of 30 faecal samples and 30 rectal swabs of each sheep and goats, 25 stool samples and 25 hand swabs of their handlers were collected from in and around Jammu district. The samples were enrichment by inoculated into MacConkey's broth tubes at 37ºC for 18-24 hours followed by selective plating. The confirmation of presumptive E. coli isolates was carried out by biochemical tests. However, the in-vitro pathogenicity was carried out by haemolysis on blood agar and by Congo red binding assay. Sixty five biochemically confirmed isolates including 27 of sheep, 26 of goats and 12 of their handlers were obtained. In-vitro virulence characterization of isolates exhibited that 55.5% of isolates of sheep and 50% isolates of goats took colour of Congo red dye. However, 25.9% and 34.6% isolates of sheep and goats, respectively were haemolytic on blood agar, but none of human isolate was found pathogenic on both of the in-vitro pathogenicity assays. Norfloxacin was found most potent drug with 84.6% sensitivity followed by Chloramphenicol (80%), Co-trimoxazole (75.3%) and Nalidixic acid (70.7%). However, 60% of E. coli isolates were resistant to Ampicillin and Amikacin. Prevalence of E. coli in both sheep and goats observed in present study was almost similar thus both animals may be equally important source for human infection particularly those who live in close proximity of these animals. Results of in-vitro pathogenicity indicate that the apparently healthy animals can also carry pathogenic strains of E. coli. Moreover, the resistance to the frequently used antibiotics indicates a need of judicious use of antibiotics in the field.