Tear production and intraocular pressures (IOPs) were determined in 38 and 102 wild Humboldt penguins (Spheniscus humboldti), respectively, from the Punta San Juan Marine Protected Area in Ica, Peru. Tear production was measured by Schirmer tear test, and IOP was measured with a TonoVet rebound tonometer. Adult (n = 90) and chick (n = 12) penguins were sampled from 2 different beaches (north and south facing) during 2 sampling years (2010 and 2011). Results showed a mean +/- SD (range) of 9 +/- 4 (2-20) mm/min for tear production and 28 +/- 9 (3-49) mm Hg for IOP. Tear production in penguins differed between beach and sex, whereas IOP differed between age, year, and beach. The IOPs were negatively correlated with packed cell volume. Tear production and IOP values had greater variation in this population than it has in other avian species. Previous investigations of IOP and tear production in Spheniscus species were conducted with birds housed under professional care in artificial marine and freshwater environments. This is the first study, to our knowledge, investigating tear production and IOP in wild penguins and establishes valuable reference intervals for this species.