BSE was confirmed in a cow in the central province of Manitoba, the Canadian Food Inspection Agency (CFIA) said.
"Final test results have confirmed bovine spongiform encephalopathy (BSE) in a mature cross-bred beef cow from Manitoba," the CFIA said in a statement.
The discovery marks the sixth time BSE has been discovered in Canada since 2003.
The disease causes brain wasting, and in its advanced stages, cattle lose the ability to walk or even stand up, displaying so-called mad behaviour.
Creutzfeldt-Jakob disease is thought to be the human form of BSE and the symptoms are similar.
CFIA officials said no part of the animal's carcass with BSE in Manitoba entered the human food or animal feed systems.
The cow was purchased with other cattle in 1992 and was born well before Canada's new feed ban to stem the spread of BSE, they said.
An investigation has been launched to identify others from the same and an offspring born in 2004, as well as to trace any feed it may have consumed at a younger age.
"Given the animals age, investigative efforts may be constrained by few surviving animals and limited sources of information, such as detailed records," officials said.
A much wider outbreak of BSE hit Britain in the late 1980s. The disease was widely controlled by banning the use of feed that included brain or spinal tissue from other cattle.