- Luigi as Miguel
- Mario as Tulio
- Princess Daisy as Chel
- Funky Kong, Wario, Waluigi, Diddy Kong, Yoshi, Princess Rosalina, Princess Peach, Chunky Kong, Cranky Kong, Toadbert, Mallow, Bommer, Fleep, Dashshell, Piccolo and Oogtar as El Dorado Villagers
- Color Toads as Big Mans
- Donkey Kong as Altivo
- Lemmy "Hip" Koopa, Iggy "Hop" Koopa and Larry "Cheasty" Koopa as Tzekel-Khan
- Bowser Jr as Cortes
- Smithy as Statue Jaguar
- Tumble as Bibo the Armadillo
- Baby Mario and Toad as Two Kids
- Candy Kong and Birdo as Two Girls in Barcelona
- Mr. L as Spanish Guard
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