The Responsible Pet Ownership bylaw was approved by council on Wednesday and will kick in at the beginning of 2015.
Cats owners will pay $15 for a spayed or neutered feline and $50 dollars for one who hasn't been spayed or neutered.
Failure to licence a cat could be subject to a $250 fine.
The bylaw also prohibits travelling shows from using exotic animals such as lions or tigers.
Bill McDonald of the Winnipeg Humane Society has said the confinement of wild animals for the circus just can't continue, saying it puts the animals under enormous stress.
Some such travelling shows have threatened to sue the city over the bylaw and Mayor Sam Katz has welcomed them to do so.