Holy basil essential oil (HBEO) can be applied as a feed additive; however, its benefits depend on the available amount in the gastrointestinal tract. In this study, the physicochemical properties, including the release properties of three different microcapsules, HBEO-loaded gelatin microcapsules (UC), UC coated with aluminum carboxymethyl cellulose (CC), and UC coated with aluminum carboxymethyl cellulose-beeswax composite (CB), were compared. The encapsulation efficiency, HBEO content, and 2,2-diphenyl-2-picrylhydrazyl radical scavenging activity for the microcapsules were 95.4 ± 0.17%, 66.7-67.7%, and 94.3-96.5%, respectively. Scanning electron microscopy and confocal laser scanning microscopy (CLSM) revealed nonuniform HBEO distributions in honeycomb-like networks in the microcapsules. An X-ray diffraction analysis determined that UC and CC microcapsules were amorphous, but CB microcapsules were semicrystalline. UV-vis spectrophotometer and CLSM analyses results determined that HBEO was released from CC and CB microcapsules in greater amounts than from UC microcapsules in simulated intestinal fluid. Therefore, the HBEO amount reaching the intestine can be controlled using the optimal encapsulation system. © 2014 American Chemical Society.