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- Mario as Thomas
- Donkey Kong as Gordon
- Rabbit as James
- Yoshi as Percy
- The Mayor of Townsville as Sir Topham Hatt
- Madame Blubbery as Alicia Botti
- Blossom and Bubbles as Annie and Clarabel (non speaking role)
- Mr. Krabs as Edward (cameo)
- Diddy Kong as Henry (cameo)
- Alec Baldwin as Narrator