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- Darkwing Duck as the Batman
- Launchpad McQuack as Robin
- Gosalyn Mallard as Batgirl
- Honker Muddlefoot as Red Robin
- J. Gander Hooter as Alfred
- Vladimir Goudenov Grizzlikof as Commissioner Gordon
- Derek Blunt as Lucius Fox
- Quackerjack as the Joker
- Negaduck as Two-Face
- Taurus Bulba as Bane
- Morgana McCawber as Catwoman
- Bushroot as Poison Ivy (it actually works)