Estrus identification is important in dairy cow production. At present, estrus identification is automated with a pedometer or accelerometer and the results remain unsatisfactory. It was previously reported that body temperature changes during estrus. In the present study, dairy cow vaginal temperature (VT) was monitored during various seasons, and an increase in VT of 0.3 °C was suggested for the onset of estrus, using an automated VT monitoring system developed in-house. Natural and synchronized estrus were measured simultaneously. The VT was determined to be in circadian rhythm and significantly higher in summer than in either autumn or winter (P < 0.05). VT difference (between estrus VT and average VT 7 days earlier) gradually increased, reached a peak of 0.56 °C ± 0.17 at 4 h before the end of estrus, and then decreased to the normal. The VT of cows in estrus and the duration of their estrus were significantly affected by seasons and estrus types (P < 0.05). VT gradually decreased in response to prostaglandin (PG) injection and was significantly lower (0.15-0.35 °C) from 9 to 33 h after the drug administration than the average VT at the same time 7 days earlier (P < 0.05). Changes in circadian and seasonal VT and in the estrous cycle can be monitored to assess the physiological status of cows and will help in developing an effective automated estrus identification technique. Results of this pilot study should be validated in further studies.