In May 2017, one large female tube-nosed bat and two small male horseshoe bats were collected in Xingyi City and Anlong County of Guizhou Province by using mist net, respectively. One of them was identified as Hairy-winged Bat (Harpiocephalus harpia), the other two were identified as Rhinolophus huananus based on the characteristics of external morphology, skull and echolocation calls, which are two new records of Chiroptera in Guizhou Province. We observed a slight bulge in the abdomen of the female H. harpia, which indicated it may be pregnant. Thus, it was released after external morphology was measured and photographed. In this case, its skull data was absent. The specimens of R. huananus were preserved in College of Environment, Northeast Normal University, China. H. harpia has short tube of the nostrils. Its pelage was thick and soft, and hind foot, wing membrane and uropatagium were covered with tawny hair (Fig. 1). The body weight is 14.01 g with forearm length 48.23 mm (Table 1). The echolocation call of this species is frequency modulation (FM) type with peak frequency 56.04 ± 4.52 kHz at resting status (Fig. 2a, Table 2). R. huananus has large ears but relatively small of the tragus. The wide noseleaf of horseshoe was completely covered its muzzle. Its sella was short (height slightly more than width) with almost rectangular, and in not being tongue shaped (Fig. 3a). The body weight of the two specimens was 4.52 g and 4.12 g, and forearm length was 40.70 mm and 40.00 mm (Table 3). The skull narrow (Fig. 3b), and the greatest length of skull was 16.35 mm and 16.46 mm (Table 3). The type of echolocation sounds is frequency modulation-constant frequency-frequency modulation (FM-CF-FM), and the peak frequency are 68.48 ± 0.08 kHz and 67.18 ± 0.06 kHz at resting status (Fig. 2b, Table 2). The discovery of two bats species expanded their distribution region in China, enriched the ecological data and provided the basic basis for further research and protection.