Manager Ron Gardenhire said Casilla had difficulty running when he tested his legs before the Twins played the Cleveland Indians.
The Twins could not get a player to Cleveland in time to fill in for Casilla, who was activated Friday but hurt his hamstring trying to beat out a bunt during Minnesota's 3-2 loss. The Twins announced that infielder Trevor Plouffe officially will be recalled from Triple-A Rochester on Sunday.
Gardenhire injected a little more levity into the situation when he said Casilla was still complaining that he beat the throw and should have been ruled safe on the bunt play.
"He was safe then, but now he's out again," Gardenhire said.
Casilla, hitting .260 with two homers and 21 RBIs in 97 games, had been sidelined since July 28 by the bad hamstring.
The Twins have had 20 different players go to the disabled list this season.