Madagascar 3: Europe’s Most Wanted took nearly $60.4 million over the weekend, according to studio estimates.
Sci-fi thriller Prometheus, starring Noomi Rapace, Michael Fassbender and Charlize Theron, debuted in second place with $50 million.
The film is rated R in the US, meaning under-17s must see it with an adult.
Madagascar 3, which premiered at Cannes Film Festival last month, reunites voice stars Ben Stiller, Chris Rock, Jada Pinkett Smith and David Schwimmer.
The Dreamworks Animation outperformed the $47.2 million debut of the 2005 original, but fell short of Madagascar: Escape 2 Africa, which took $63.1 million in its opening weekend.
The third instalment, which has been a bigger hit with the critics, benefited from higher ticket prices and more expensive 3D shows.
Prometheus, telling the story of a team of explorers who discover a clue to the origins of mankind, was also screened in 3D.
Last weekend’s US number one Snow White & the Huntsman slipped to third-place, with Men in Black 3 at number four.
Fifth place went to The Avengers, which has now taken $571.9 million at the US box office – making it the highest-grossing film of the year so far.
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