The pattern of antimicrobial resistance (AMR) and the prevalence of multi drug resistance (MDR) of Escherichia coli (E. coli) isolates from livestock, wild animals and an ornamental fish farm were investigated in a selected area in Polgahawela, Sri Lanka. Sampling area of one square Km containing an ornamental fish farm, a broiler farm, cattle, goat, and backyard poultry which are located separately within the sampling area but not in an integrated farming system with its associated wildlife was mapped using GPS. Faecal samples were examined from 89 wild animals (terrestrial and avian species), 28 livestock animals (including poultry) as well as 40 samples from an ornamental fish farm (fish faecal material, pond water and pond sediment). E. coli was isolated using standard methods and AMR profiles were investigated for 12 antimicrobials using Kirby-Bauer disc diffusion method. Sixty nine percent (62/89) of wild animals and 86% (24/28) of livestock samples were positive for E. coli whereas the isolation rate was 17.5% (7/40) in the samples from the ornamental fish farm. A maximum of two randomly selected E. coli isolates from each sample, making a total of 164 isolates were tested for AMR of which 114, 43 and 7 were from wild animals, livestock and the ornamental fish farm, respectively. The E. coli isolates tested against 12 antimicrobials reflected that the proportions of E. coli isolates resistant to at least one antimicrobial were 30%, 63% and 71% in wild animals, livestock and the ornamental fish farm, respectively. Multidrug-resistant E. coli isolates were detected in wild animals, livestock and the samples from ornamental fish farm. Overall, according to the statistical analysis, E. coli isolated from livestock and the ornamental fish farm reflected a significantly higher chance of carrying AMR compared with E. coli from wild animals.