In this study, we investigated the effect of royal jelly on the puberty of female 1/4 Romanov x 3/4 Morkaraman crossbred lambs. Royal jelly was applied in two ways: orally and intravaginally. The rate of estrus as well as the day and time (in hours) of estrus were observed and determined by real-time ultrasonography (for pregnancy) and laparoscopic surgery (ovary examination). Three groups of lambs were observed. The first group, consisting of 15 female lambs, was treated intravaginally; in the second group, 15 female lambs were treated orally; and the third group, which served as control, consisted of 10 ewe lambs that were not treated with royal jelly. Estrus rates in the intravaginally and orally treated groups were 13% and 20%, respectively, whereas the control group showed no estrus activity. However, estrus interval in both groups was observed 3-5 days after the withdrawal of the treatment. All the animals had an active ovary when observed during estrus activity by ovary examination. Yearling 1/4 Romanov-bred ewes respond positively to royal jelly application. Increasing the doses and extending the duration of royal jelly application might yield better results.