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New enemies
| Image | Description | Location |
|---|---|---|
 Amps |
A giant Bob-omb in Bob-omb Battlefield. He appears as the first boss in the game and has the first Power Star. | Bob-omb Battlefield |
 Bookends |
A giant Whomp on top of Whomp's Fortress. He has the first Power Star, and after he is defeated, there is a tower on top of the fortress. | Whomp's Fortress |
 Bubbas |
A giant Boo that appears three times in Big Boo's Haunt. He initially appears once all the Boos in the mansion have been defeated, again in the underground Merry-Go-Round, and lastly on the top balcony of the mansion. | Big Boo's Haunt |
 Bullies |
A giant Mr. I which only appears in the attic of Big Boo's Haunt. | |
 Butterflies |
Bowser appears in Bowser in the Dark World, Bowser in the Fire Sea (where the arena tilts), and Bowser in the Sky (where he must be thrown three times). Mario needs to swing him by the tail clockwise or counterclockwise and hurl him at bombs on the outside of his circular arena. | Bowser in the Dark World Bowser in the Fire Sea Bowser in the Sky |
 Chairs |
Two of these giant Bullies appear in Lethal Lava Land. On both occasions, they try to knock Mario into the lava. | Lethal Lava Land |
 Chuckyas |
A giant Wiggler, which becomes angry when his home in Tiny-Huge Island gets flooded. | Tiny-Huge Island |
Returning enemies
| Image | Description | Location |
|---|---|---|
 Beach Koopas |
A giant Bob-omb in Bob-omb Battlefield. He appears as the first boss in the game and has the first Power Star. | Bob-omb Battlefield |
 Big Goombas |
A giant Whomp on top of Whomp's Fortress. He has the first Power Star, and after he is defeated, there is a tower on top of the fortress. | Whomp's Fortress |
 Big Steelies |
A giant Boo that appears three times in Big Boo's Haunt. He initially appears once all the Boos in the mansion have been defeated, again in the underground Merry-Go-Round, and lastly on the top balcony of the mansion. | Big Boo's Haunt |
| Bullies |
A giant Mr. I which only appears in the attic of Big Boo's Haunt. | |
 Bob-ombs |
Bowser appears in Bowser in the Dark World, Bowser in the Fire Sea (where the arena tilts), and Bowser in the Sky (where he must be thrown three times). Mario needs to swing him by the tail clockwise or counterclockwise and hurl him at bombs on the outside of his circular arena. | Bowser in the Dark World Bowser in the Fire Sea Bowser in the Sky |
 Boos |
Two of these giant Bullies appear in Lethal Lava Land. On both occasions, they try to knock Mario into the lava. | Lethal Lava Land |
 Bubs |
Two stone hands with eyes on their palms. They appear when Mario blasts open the top of the Pyramid in Shifting Sand Land and rides the elevator inside. | Shifting Sand Land |