The aim of the present study was to determine the stages of the reproductive cycle of the bitch by using the exfoliative vaginal cytology technique and its relationship with hormonal level. A total of 12 bitches (n=12) were selected and vaginal cytology along with hormonal estimation was done to identify the different stages of the reproductive cycle. The vaginal samples were collected as per the cotton swab technique and stained with Leishman stain and examined microscopically for differential count of exfoliated vaginal epithelial cells. On the same day blood samples were also collected aseptically for hormonal estimation. The percentage of parabasal cell, intermediate cell and superficial cells were 7.04±0.28, 22.33±0.28, and 62.32±0.28 in proestrus; 0.56±0.03, 5.68±0.03 and 92.29±0.03in estrus and 32.56±0.28, 35.08±0.28 and 12.83±0.28 in diestrus respectively. Vaginal smears were characterized by presence of massive numbers of erythrocytes and disappearance of neutrophils in proestrus, clear background of smear and absence of neutrophils with scanty erythrocytes in estrus, infiltration of neutrophils and dirty background in diestrus. The estrogen level (29.50 ±0.28) was statistically high during proestrus and highest progesterone level (39.50 ±0.28) during diestrus. On conclusion, hormonal changes in different stages of reproductive cycle presented mainly by the effect of elevated estrogen hormone that associated with increase in superficial cells count and cornification, with reduction in neutrophil and parabasal cell in estrus and Progesterone hormone showed increased towards diestrus level with increasing intermediate cells.