The NBC show came to Toronto last week with the help of $750,000 in Canadian taxpayers' money, a trip that Canadian promoters hoped would help rebuild the city's tourist industry after an outbreak of severe acute respiratory syndrome last year.
In the pre-taped skit that aired Thursday night, Triumph the Insult Comic Dog confronted visitors to Quebec City's Winter Carnival.
When one couple confirmed they were French Canadian separatists, the grouchy puppet said, "Listen closely. Hear that? It's the sound of no one giving a," followed by an expletive bleep.
"You're in North America, learn the language!" he hollered at another.
Triumph asked a rotund man if he was a separatist, then suggested he might want to separate himself from doughnuts for a while. The skit also replaced street signs with insulting replicas, including one that mocked Canadian singer Celine Dion.
Mauril Belanger, the deputy government house leader, told the House of Commons Friday that the government was not amused. He said Ottawa completely dissociated itself from the material.
Alexa McDonough, a member of Parliament for the New Democratic Party, said the sketch was "vile and vicious" and amounted to hate-mongering. McDonough said the government should demand its money back.
"The whole point of trying to help deal with the devastation of the SARS crisis on the city of Toronto was to attract tourists," McDonough said. "How it got morphed into this kind of garbage I don't know."
Chum Television, which airs the show in Canada, issued an apology and removed the segment in a repeat broadcast.
"It is never our intention to air programming that offends any of our viewers," the network said.
NBC has declined comment, and O'Brien mentioned the topic only in passing in his monologue during the taping of the final Toronto show on Friday afternoon.
Guest Jim Carrey, a Canadian, later offered O'Brien a subtle opportunity to apologize by bringing on the mascot for the winter carnival, but O'Brien didn't take the bait.
Separatism and French language rights are a sensitive issue in Canada, especially after Quebec separatists nearly won a 1995 referendum to set up their own country.
Canada's current government is immersed in a scandal over allegations that Quebec advertising agencies were paid $75 million in illegal commissions to confront the separatist threat.
"It's wrong to spread the worst possible ethnic prejudices about Quebecers, which already exist in Canada," said Jean Dorion, president of the Quebec nationalist Saint-Jean-Baptiste Society.
But businessman Peter Soumalias, said who helped bring the New York-based show to Toronto, said Canadians were taking the show too seriously.
"It's a silly puppet that tells silly jokes," Soumalias said. "Most people find it funny."