Aims: To evaluate an alginate-chitosan microcapsule for an intestinal probiotic delivery system for broilers. Methods and Results: Lactobacillus reuteri KUB-AC5 was successfully microencapsulated with alginate and chitosan mixtures using an emulsion cross-linking method with high microencapsulation efficiency. Scanning electron microscopy revealed a large number of the bacteria entrapped in the semi-interpenetrating network. The microcapsule effectively protected the cells against strong acids. The in vitro study showed that the 8 log CFU g(-1) was released at the jejunum and ileum. For the in vivo study, the number of probiotics was detected by a polymerase chain reaction-based random amplified polymorphic DNA technique. From provision of 10 log CFU, cell numbers of 5-8 log CFU were observed in the intestine. Conclusions: The alginate-chitosan microcapsule can serve as a potential intestine-targeted probiotic delivery system. Significance and Impacts of the Study: To the best of our knowledge, this is the first comparison study of the in vitro and in vivo gastrointestinal profiles of microencapsulated probiotics used as feed additives for broilers. This study reveals the similarities and differences of the in vitro and in vivo probiotic profiles and provides convincing evidence of the potential use of the alginate-chitosan microcapsule as a probiotic delivery system.