Queenless, caged, newly emerged worker bees (Apis mellifera L.) were fed honey, 22 and 40% pollen in honey, and 22 and 40% royal jelly in honey for 14 days. Workers fed royal jelly, pollen, and honey had large, medium, and small ovaries, respectively. Royal jelly had higher nutritive value for workers’ ovarian development than did pollen, possibly because royal jelly is predigested by nurse bees and easily used by adult and larval bees. These results suggest that nurse bees could mediate workers’ ovarian development in colonies via trophallactic exchange of royal jelly. Six levels of royal jelly in honey, 0, 20, 40, 60, 80, and 100% (royal jelly without honey), were tested for their effects on workers’ ovarian development and mortality for 10 days. High levels of royal jelly increased ovarian development, but also increased worker mortality. All caged bees treated with 100% royal jelly died within 3 days. When workers were incubated at 20, 27, and 34 °C for 10 days, only bees at 34 °C developed ovaries. These findings suggest that nurse bees functioning as units which digest pollen and produce royal jelly may feed some potentially egg-laying workers in a brood chamber with royal jelly when a queen is lost in a colony. Feeding workers a diet of 50% royal jelly in honey and incubating at 34 °C for 10 days is recommended for tests of ovarian development.