The current study was designed to evaluate the changes in vaginal cytology and serum progesterone and estrogenةconcentration in anestrum bitches treated with a combination of FSH and LH. Seven healthy Mongrel anestrum bitches were treated with daily administration of 75 I.U. FSH plus 75 I.U LH for consecutive nine days and another three bitches were kept as a control without any treatment. Treated and control bitches were examined for the percentages of different vaginal-cytology cells, serum concentration of estrogen and progesterone, and signs of proestrus and estrus in a scheduled timeline. Five of the seven treated bitches (71.4%) showed a gradual decrease in the percentage of parabasal cells, a gradual increase in the percentage of cornified and RBCs, a rapid decrease in the estrogen concentration, and an increase in the progesterone concentration. These five bitches showed signs of proestrus and four of them showed male acceptance and were conceived. These five bitches had high estrogen concentrations before initiation of the treatment. The other two bitches had a low estrogen concentration before the initiation of the treatment and after treatment, they showed a gradual increase in the estrogen concentration, but they did not show characteristic changes in vaginal cytology or progesterone concentration. Control bitches had high estrogen concentrations before treatment, but they did not show characteristic changes in vaginal cytology, estrogen and progesterone concentrations, or signs of proestrus up to one month after the end of the treatment. In conclusion, treating anestrum bitches with a mixture of FSH and LH induced characteristic changes in vaginal cytology and serum estrogen and progesterone concentrations in the majority of bitches. These changes were associated with the induction of fertile estrus in more than half of the treated bitches, and this percentage increased to about 80% in bitches having initial high estrogen concentration.