Abstract: Background: Escherichia coli is one of the most common causative agent of bacterial diseases. The prevalence of E. coli infection and its outbreak has been reported in many countries but scanty information is available on the antibiogram of this bacterium in Cape Coast, a major tourist destination in West Africa. Materials and Methods: The antibiotic susceptibility and occurrence of Escherichia coli isolated from clinical and environmental samples from the Cape Coast Metropolis were investigated. Bacteria isolation and identification were carried out using various bacteriological media and Analytical Profile Index (API) 20 E kits, respectively. All the test E. coli isolates were screened for their susceptibility to 16 antibiotics. Results: In all, 389 E. coli isolates were obtained comprising 261 and 128 from clinical and environmental samples respectively. All E. coli isolates were 100% sensitive to Imipenem. The percentage sensitivities of clinical E. coli isolates to ampicillin (0-24.1%), tetracycline (16.0-28.4%), cotrimoxazole (16.8-22.0%), cefuroxime (27.6-43.2%) and nalidixic acid (22.1-47.8%) were found to be relatively low. The sensitivity of environmental isolates to the 16 antibiotics was higher than that of clinical isolates, except for nalidixic acid, aztreonam and amikacin to which isolates from environmental samples were less sensitive. Among clinical samples, stool samples had the highest percentage occurrence of E. coli (44.4%), followed by urine samples (36.4%); while blood had the lowest percentage (1.6%). Among environmental samples, the highest percentage occurrence of E. coli (44.2%) was recorded for the Fosu lagoon, followed by fresh beef samples (21.8%) and fresh chicken (4.7%). No isolates were obtained from seawater, cabbage and smoked fish. Conclusion: Escherichia coli isolated from environmental samples were more sensitive to most antibiotics used in this study compared to clinical isolates. This study has demonstrated that environmental samples may harbour some considerable level of antibiotic resistance and hence underscores the need to dispose off waste properly.