The aim of this study was to compare the appearance rate of vaginal cytology during estrous cycle in small pet bitches. A characteristic features of vaginal cytology during the estrous cycle were the high proportion of superficial cell in proestrus, anuclear cell in estrus, small intermediate cell in diestrus, and small intermediate cell in anestrus, respectively. There were no statistically significant differences of appearance rate of vaginal cytology among small pet bitches in the each phase of estrous cycle. These results indicated that the vaginal cytology was useful method for estimating estrous cycle and optimal breeding time in small pet bitches.