The aim of this study was to determine the stages of the sexual cycle of the bitch by direct examination and also to assess the reliability of this new technique by comparing it with the classical staining techniques used in bitches. Forty mixed-breed bitches, of different ages and sexually mature were used in this study. A total of 120 vaginal smear samples were collected using a cotton swab, three from each bitch. The collected samples were air dried and coded. One of the prepared samples from each bitch was stained with May-Gnmwald Giemsa and the second sample was stained with Papanicolaou. The third sample was left unstained for direct examination. The stages of the sexual cycle were determined using all of the samples. The researcher who evaluated the samples did not have information about the coding system. The evaluation was made blindly and the results were compared after determination of the stages of the sexual cycle from all of the samples. The sexual cycle stages determined with the May-Gnmwald Giemsa and Papanicolaou techniques were completely consistent with each other. However, when the direct examination technique was compared with the classical staining techniques, there was a significant difference in the proestrus, diestrus and anestrus stages of the cycle (p<0.05) while there was no significant difference (p>0.05) in the estrus stage of the cycle. In conclusion, it was found that when determining the stages of the sexual cycle of the bitch by vaginal cytology, the direct examination technique was reliable only in detecting the estrus stage of the cycle. © Medwell Journals, 2011.