Five healthy captive greater one-horned rhinoceroses (Rhinoceros unicornis) were evaluated with standing sedation (detomidine hydrochloride 20–30 mg and butorphanol tartrate 50–60 mg per animal) to determine baseline values for tear production, intraocular pressure (IOP), conjunctival flora, and ocular anatomy using brightness mode transpalpebral ultrasonography with a 4- to 7-MHz broadband curvilinear transducer. The mean Schirmer tear test I value was 18.2 ± 3.49 mm/min. The mean IOP measured using applantation tonometry was 31.2 ± 6.62 mm Hg. Ocular biometry measurements were the following: axial length 2.61 ± 0.11 cm; corneal thickness 0.13 ± 0.01 cm; anterior segment depth 0.28 ± 0.06 cm; lens depth 0.70 ± 0.11 cm; and posterior segment depth 1.46 ± 0.13 cm. These values indicate that the globe is smaller than that of the domestic horse. All eight conjunctival swabs cultured bacterial and fungal microorganisms, with the most common being Staphylococcus spp. (57%). All bacterial isolates were considered to be commensal organisms due to the presence of mixed bacterial populations and lack of clinical signs of ocular disease. The data collected in this study should provide veterinarians with baseline information to assist in the diagnosis of ophthalmic conditions in the greater one-horned rhinoceros.