We conducted a species diversity investigation for Chiroptera (bats) in Guangdong Shimentai National Nature Reserve from 2017 to 2019 by using daily/night roost surveying and mist-net capturing method. Total of 31 species (4 families, 9 genera) were investigated (Table 1). Of the 31 species, 7 (22.6%, 1 genus) are from Rhinolophidae, 4 (12.9%, 1 genus) from Hipposideridae, 1 (3.2%, 1 genus) from Megadermatidae and 19 (61.3%, 6 genera) from Vespertilionidae. Of these species, 19 are the Oriental type, which accounts for the largest proportion. The rest species, seven belong to South China type, and five belong to monsoon type. Bats from this survey in Shimentai National Nature Reserve were roosted in cave, the rest species either in larger trees or in buildings. Of them, 12 species are belonged to cave-dwelling, 4 building-dwelling, and 4 arboreal-dwelling. In addition, 8 species live in both caves and buildings, and 3 species inhabit both buildings and trees. As Guangdong Shimentai National Nature Reserve covers a relatively large area, this reserve is one of the reserves with rich bat biodiversity. This survey has investigated the data of species diversity of bats in Guangdong Shimentai National Nature Reserve, which provides the basic data for the long-term study of the widelife resources.