Objective: The clinical uses of probiotics are increasing. As medical students are our future physicians, their knowledge and attitude towards probiotics will dictate the extent to which probiotics are prescribed in the future. We investigated medical students’ knowledge and attitudes towards probiotics during their clinical years (4-6 years) at the Jordan University of Science and Technology, Irbid, Jordan. Methods: This is a cross-sectional cohort study using online/printed questionnaire collecting personal and socio-demographic data in addition to students’ knowledge and attitude towards probiotics, the clinical usage, and availability of probiotics, and students’ source of information on probiotics and whether probiotics are covered in their curriculum. We presented their responses as averages and compared these by gender and lifestyle. Results: Total 272 (15%) students completed the questionnaires (144 females, 22.93 ± 1.13 years old), 140 (51.5%) of which claimed to have a healthy lifestyle. More than half the students could correctly define probiotics, knew the sources and could identify at least 2 usages; such knowledge did not differ by gender. Most students 225 (82.3%) recognized the mechanisms of action of probiotics, but only 115 (42.2%) knew that probiotics’ effects are strain-specific. Most students (244 (89.7%)) had a positive attitude towards probiotics and were interested in learning more about probiotics (252 (92.6%)). Their attitudes did not differ by lifestyle (p=0.196). Conclusion: Medical students in Jordan had an acceptable level of knowledge about probiotics and their health effects. Covering probiotics in medical school curricula may boost students’ knowledge, thereby enhancing future prescription of probiotics.