The goals of this study were to compare ocular morphology, determine the reference intervals of selected ophthalmic tests, ocular measurements, intraocular pressure, and tear production, and to establish possible relationships in the visual ecology of three different Neotropical nonhuman primates (NHP). Nineteen black-tufted marmosets (Callithrix penicillate), 24 Guianan squirrel monkeys (Saimiri sciureus), and 24 night monkeys (Aotus azarae infulatus) were included in the study. Schirmer tear test, ocular dimensions, ocular ultrasonography, intraocular pressure, central corneal thickness, and corneal touch threshold were determined. The ratio of the average corneal diameters and axial diameters (CD/AGL) were established. No significant difference was noted between males and females, nor left and right eyes, for all three species for all measurements (P > 0.05). CD/AGL ratio was significantly higher (P < 0.0001) in night monkeys (a nocturnal species) as compared to black-tufted marmoset and Guianan squirrel monkeys (two diurnal species). The reference intervals will aid veterinary ophthalmologists to more accurately diagnose pathological changes in the eyes of these species. In addition, ocular dimension comparison will allow other NHP species to be evaluated and examined in relationship to behavioral traits (nocturnal versus diurnal).