This study was designed to evaluate the ability of mice to discriminate cow urinary odor from different reproductive phases, with a view toward detecting the estrous phase. Experiments were also carried out to establish the relationship between androgen and mouse behaviors during estrous detection. Further, the study was also intended to establish the relationship between androgen and behaviors in estrous detection. Bovine urine was collected during estrus and non-estrous periods, i.e., prepubertal, preovulatory, ovulatory, postovulatory, pregnancy, and lactation. Behavioral analyses were carried out in a Y-maze apparatus, in which the mice were acclimatized in before odor-preference tests. The number and duration of visits, and grooming behavior by male responders towards the urine samples, were recorded. Intact male mice showed a higher response towards estrus urine samples than towards non-estrous urine. By contrast, orchidectomized mice failed to discriminate estrous urine, whereas castrated mice treated with testosterone regained the ability to discriminate estrus odor. A higher level of grooming behavior was found in males exposed to estrous urine than to urine of other phases. These results suggest that normal mice have the ability to detect estrus, and that this discriminating ability is androgen dependent. The grooming behavior shown by males in response to estrous urine may be taken as a key parameters in estrous detection. The results further suggest that bovine estrous urine produces specific odors that probably involve both intraspecific and interspecific communication.