According to officials at Animal Ed. Ventures, the otter enclosure and sanctuary gates were found open Tuesday morning.
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The three otters, Sigmund, Nessy and Ned, were all missing.
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The gates were double-checked prior to closing on Sunday night, according to the sanctuary director Cory Freeman.
"The latch to their enclosure was completely intact," she said. "There was no failure. This is a latch that can only be manipulated manually with a level of dexterity."
She said they also heard a loud high-pitched noise around 2 a.m.
Freeman isn't sure why someone would steal the otters.
"We don't know if this is motivated by money, we don't know if this was motivated by an extremist group that feels like all animals should be free," she said. "Are they being properly cared for? Does this person even know what they eat and what they need? They're not a creature that you can just put in a cage.... It's devastating."
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The sanctuary is home to more than 200 animals that were once kept as pets.
Anyone with information is urged to contact Detective Gardner at Harnett County Sheriff's Office (910)893-9111 or Animal Ed.Ventures at email, sanctuary@animal-edventures.org.