Probiotics have been suggested as an effective adjuvant treatment for Alzheimer’s disease (AD) due to their modulating effect on the gut microbiota, which may affect the gut-brain axis. Therefore, we aimed to evaluate the effects of two different single-strain probiotics on cognition, physical activity, and anxiety in subjects with mild and moderate AD. Eligible patients ( n = 90) with AD were randomly assigned to either of two interventions [ Lactobacillus rhamnosus HA-114 (10 15 CFU) or Bifidobacterium longum R0175 (10 15 CFU)] or placebo group, receiving probiotic supplement twice daily for 12 weeks. The primary outcome of the study was cognitive function measured by using the two tests, namely, the Mini-Mental State Examination (MMSE) and the categorical verbal fluency test (CFT). Secondary outcomes included a performance in Activities of Daily Living (ADL), the Lawton Instrumental Activities of Daily Living (IADL) scale, and the Generalized Anxiety Disorder (GAD-7) scale. Linear mixed-effect models were used to investigate the independent effects of probiotics on clinical outcomes. After 12 weeks, MMSE significantly improved cognition ( P Interaction < 0.0001), with post hoc comparisons identifying significantly more improvement in the B. longum intervention group (differences: 4.86, 95% CI: 3.91–5.81; P < 0.0001) compared with both the placebo and L. rhamnosus intervention groups (differences: 4.06, 95% CI: 3.11–5.01; P < 0.0001). There was no significant difference between the two intervention groups (differences: −0.8, 95% CI: −1.74 to 0.14; P = 0.09). In conclusion, this trial demonstrated that 12-week probiotic supplementation compared with placebo had beneficial effects on the cognition status of patients with AD.