Two males of barbastelle were obtained near of Zhuping Village and Xiaoxi Village, Xiaoxi Town, Yongshun County, Hunan Province, on April 6th, 2014 and August 17th, 2015. It was a medium-sized species, and had triangular tragi and nearly square outline of ears with transverse ridges in shape. The ears joined across its forehead without a prominent projecting lobe, and the tragi length was nearly a half of ear (Fig. 1a). The dorsal and ventral fur appeared black brown with the hair tip slightly white, the ventral hair between patagium base and uropatagium was partially yellow-white. The skull is slightly flattened, and the small of tympanic bulla, The dental formula of specimens is 2.1.2.3/3.1.2.3 = 34 (Fig. 1b﹣e). They were identified as Asian Barbastelle (Barbastella leucomelas) based on its external morphology, skull characteristics, and the comparison of Cyt b and ND1 gene sequences by construction of Neighbor-Joining phylogenetic tree (Fig. 2). The specimens of B. leucomelas were a new record of Chiroptera for Hunan Province, preserved in the Vertebrate Collection, College of Biology and Environmental Sciences of Jishou University and Guangdong Institute of Applied Biological Resources, respectively.