This study aimed to determine species-specific reference values for intraocular pressure (IOP) in healthy gazelles (Gazella subgutturosa) living in the Şanlıurfa region by performing IOP measurements using rebound tonometry without sedation and in a natural standing posture. A total of 20 healthy individuals aged between 0 and 6 months (10 females and 10 males) were included in the study, and both eyes of each subject were evaluated separately, resulting in a total of 40 eyes. IOP measurements were performed using the Icare® TONOVET Plus device in "d" mode, without the application of topical anesthetics or sedatives, with the animals in a standing position. For each eye, the mean of six consecutive measurements was used. Data were analyzed using parametric methods, and the level of significance was set at p<0.05. The mean IOP values for the right and left eyes were 20.5±8.1 mmHg and 22.1±9.7 mmHg, respectively, with no statistically significant difference between them (p=0.471). The overall mean IOP was calculated as 21.3±9.0 mmHg. IOP was found to be significantly higher in female individuals compared to males (24.3±9.1 vs. 17.6±6.8 mmHg; p=0.026). No statistically significant difference in IOP was observed among the age groups. This study defines the first IOP reference values obtained without sedation using rebound tonometry in Gazella subgutturosa and provides clinically valuable reference data for the diagnosis and monitoring of traumatic, inflammatory, and glaucoma-like ophthalmic conditions.