There is paucity of surveillance studies on antibiotic resistance in commensal Escherichia coli. The main aim of this cross-sectional study was to determine factors associated with peri-anal carriage of multi-resistant E. coli by healthy postmenopausal women. A structured questionnaire was completed for consecutive healthy postmenopausal women aged 45 y and above attending the outpatient clinics of the Department of Obstetrics and Gynaecology at R.D. Gardi Medical College, Ujjain, India. Antibiotic susceptibility testing was done by the Kirby-Bauer disk diffusion method, as recommended in the Clinical and Laboratory Standards Institute guidelines. The study showed that 28% of healthy women carried multidrug-resistant (MDR) E. coli in the peri-anal region. The factors significantly associated with carriage of MDR E. coli were family size > 10 (OR 8.23, 95% CI 2.73-24.73; p < 0.001), antibiotic use in the past 2 weeks (OR 5.39, 95% CI 2.36-12.34; p < 0.001), and hospitalization in the past 2 weeks (OR 4.02, 95% CI 1.73-9.31; p = 0.001). This is the first study from India reporting the MDR E. coli carriage rate in healthy women aged over 45 y.