The present study aims to evaluate the awareness of a sample of the general population and health-care professionals regarding the use of antibiotics and antibiotic resistance and identify the factors associated with antibiotic resistance. Methods: This prospective questionnaire-based study was conducted for 3 months, from July to September 2020, in Karaikal, Puducherry (India). The health professionals included doctors in pre- and paraclinical teaching departments (those not involved in clinical practice), laboratory technicians, and PhD students pharmacists. The questionnaire had two parts which health professionals answered, whereas the general population answered only the second part. Findings: About 38.5% of the population reported using antibiotics frequently among the responses obtained. Around 66.5% of the general population usually stop the antibiotic within 1 or 2 days after they feel better, and 11% of the people believed that adding an extra antibiotic would make them better. Among the health professionals, more than 90% responded that self-medication is the major reason for antibiotic resistance and was unsure of the overprescription of antibiotics. Conclusion: The results show variable responses and suggest the need for intervention programs to increase the knowledge among the general population regarding the rational use of antibiotics.