- Stan as Mario
- Ollie as Luigi
- Ginger Hammond as Princess Rosalina
- Dan Forrester as Toadsworth
- Sgt. Hippo as Toad
- Colonel Ridley as Diddy Kong
- Dr. Schickel as Bowser
- Captain Baker as Donkey Kong
- Aunt Martha as Princess Daisy
- Aunt Agatha as Princess Peach
- General Talylor as Wario
- Doctor as Yoshi
- Colonel Wayburn as Oogtar
- General Burns as Bowser Jr.
- Postman as Detective Herlock Solmes
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