• Dr. John Dolittle as Mario
  • Grandpa Archer Dolittle as Yoshi
  • Dr. Mark Weller as Luigi
  • Calloway as Toad
  • Lucky the Dog as Oogtar
  • Rodney the Guinea Pig as Detective Herlock Solmes
  • Dr. Gene "Geno" Reiss as Wario
  • Lisa Dolittle as Princess Daisy
  • Dr. Fish as Waluigi
  • Rat #1 as Bowser
  • Rat #2 as Donkey Kong
  • Old Beagle as Bowser Jr.
  • Baby Tiger as Ludwig Von Koopa
  • Maya Dolittle as Princess Peach
  • Charisse Dolittle as Princess Rosalina
  • Possum as Diddy Kong
  • Bettleheim the Cat as Morton Koopa, Jr.
  • Male Pigeon as Boo
  • and more

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