The use of gut biotics, including probiotics, prebiotics, and synbiotics, has shown substantial potential in the management of various health conditions possibly through the gut-organ axis. The role of gut biotics in modulating the gut-brain axis is becoming evidentwith more research focusing on this intervention. Improvement of gut-organ axis function is possible by using food-related products called gut biotics. However, there is limited comprehension of the knowledge and use of these intestinal or gut biotics. Our aim was to recognize knowledge gaps and assess the improvement of understanding following an education intervention. Asingle-arm studyencompassing a convenient sample of 161 inpatient and outpatient subjects aged 50 years and older was conducted at the University of Alberta Hospital from June to August 2023. Knowledge about gut biotics was evaluated using a structured questionnaire consisting of 16 questions and involving six thematic areas. To ensure validity, the questionnaire was pre-tested on 10 physicians and residents who were not part ofthe study.The questionnaire was administered to study subjects prior to receiving an information sheet about gut biotics.Two weeks after receiving the information sheet, all participants were contacted by phone, and the same questionnaire was administered again. Of the 161 patients, 122 completed the pre-intervention and post-intervention questionnaires and were considered in the analysis. The mean age of the participants was 72 years (SD: 10.8), 57% comprised women, and 39% had less than a high school education. The proportion of polypharmacy and multimorbidity was87% and 97%, respectively. Following the intervention, there was a noticeable enhancement in knowledge across all the themes, with statistical significance (p<0.001) observed in 14 out of 16 questions as determined by the homogeneity statistical test. Knowledge gaps in gut biotics were prevalent among study participants, and the educational intervention effectively contributed to the enhancement of knowledge. The results of this study provide valuable information for the development of targeted health education strategies focusing on gut biotics, which may play a role in improving gut-organ axis function.