The Calgary Zoo says seven of its Humboldt penguins drowned in their holding area.
The facility said the birds were found dead Thursday morning.
Necropsies on the birds determined that they had drowned.
The birds appear in the zoo's "Penguin Plunge" exhibit alongside the king, gentoo, and rockhopper penguin species.
"This is devastating news", said Jamie Dorgan, the zoo's director of animal Care. What led them to drown is unclear.
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"We are thinking a lot of what the possibilities could be".
A British veterinary surgeon who visited the zoo after the 2013 penguin deaths told the Calgary Herald that such panic deaths can also occur in the wild. "We have humans looking after animals and just like humans doing anything, mistakes are going to happen", Dorgan says. "You know, all the animals at the zoo are family members for our staff".
Dorgan said staffers at the zoo were heart-broken over the deaths of the penguins. "We're obviously very shocked at what happened and a lot of people were very upset".
Zoo officials say the colony of 22 Humboldt penguins was fine when a zookeeper left on Wednesday afternoon.