Purpose: To determine the magnitude of ocular hypotony and the length of recovery time to 6 and 10 mm Hg IOP following anterior chamber (AC) cannulation.Methods: Bilateral IOP was recorded 500 times per second via telemetry immediately before, during, and immediately after AC cannulation with a 27-G needle in 10 different sessions at least 2 weeks apart in four male rhesus macaques (nonhuman primates; NHPs) aged 3- to 6-years old. Bilateral IOP was recorded continuously using a proven telemetry system while the NHPs were under general anesthesia during IOP transducer calibration experiments involving manometric control of IOP via AC cannulation, then continuously after the AC needles were removed until IOP recovered to precannulation levels. The change in IOP from baseline to AC cannulation was tested using the signed-rank test. The times necessary for IOP to recover to 6 and 10 mm Hg, respectively, were calculated.Results: Average precannulation IOP was 11.5 mm Hg and significantly decreased to an average of 2.3 mm Hg immediately following AC needle removal (P = 0.0156). On average, IOP recovered from 2.3 to 6 and 10 mm Hg in 32.4 and 63.7 minutes, respectively. Recovery times of IOP were not affected by repeated AC cannulations every 2 weeks.Conclusions: Generally, IOP recovers relatively quickly after repeated AC cannulation, and did not result in extended duration hypotony. It is important to consider hypotony in animal experiments and clinical procedures involving AC cannulation and paracentesis when consideration of IOP or its effects is important.