Two bats (sample number are HUNNU16SF16 male and HUNNU16SF38 female) have been captured by mist net in Xianfei Cave (26°58′25″N, 113°3′23″E, altitude 463 m), Sifang Mountain, Hengdong County, Hunan Province. The bats were identified as Myotis petax based on its external morphology, skull characteristics and molecular data. The measurement of forearm length was 40.73 mm and 42.40 mm for male and female which is slightly longer than that of M. petax captured from other locations (Table 2). The baculum Length in M. petax is 1.088 mm, nearly twice as large as in M. daubentonii. Maximum Likelihood tree based on Cyt b sequences was constructed (Fig. 4). The cladogram indicated the sequence of the samples from Hengdong County form a clade with M. petax that collected from Jilin Province China, Russia and Japan. The bat is a new record of Chiroptera in Hunan province. The samples were preserved in Hunan Normal University Vertebrate Specimens Museum.