To establish reference values for ophthalmic tests in two bat species. BATS: Fourteen bats including seven great fruit-eating bats (Artibeus lituratus) and seven tailed tailless bats (Anoura caudifer). Normal values for following ophthalmic tests were investigated as follows: (a) aqueous tear production using the standardized endodontic paper point tear test (EPPTT), (b) rebound tonometry, and (c) horizontal palpebral fissure length. Aqueous tear production was 2.53±1.65mm/min for Alituratus and 1.89±0.62 for Acaudifer. Intraocular pressure measured in the upright position was 11.0±3.28mmHg for Alituratus and 7.28±2.70 for Acaudifer. Horizontal palpebral fissure length was 5.04±0.45mm for Alituratus and 3.92±0.51 for Acaudifer. The data obtained in the present study may serve as a reference for ophthalmic parameters and help practitioners in the diagnosis and management of eye diseases in bats, as well for future investigations about microchiroptera bats.