The two top keepers are both injured and striker Fakhreddine Ben Youssef will act as Mathlouthi's back-up after FIFA rejected Tunisia's request to add a replacement keeper to the squad.
Defenders Dylan Bronn and Syam Ben Youssef were also ruled out due to injuries suffered against Belgium.
Panama coach Hernan Dario Gomez made four changes to the side that lost 6-1 to England, with midfielder Armando Cooper and defender Michael Murillo missing out through suspension.
Defenders Adolfo Machado and Luis Ovalle come into the side, along with striker Gabriel Torres and midfielder Ricardo Avila.
Neither team can reach the knockout phase, with England and Belgium meeting in Kaliningrad in the other Group G match to decide the two top places.
(Reporting by Angus MacSwan; Editing by Ed Osmond)
