Centre-back Benatia did not feature at the weekend and has a knee problem.
Neither player has travelled with Massimiliano Allegri's squad for Tuesday's Group H match, where victory at the Ramon Sanchez Pizjuan would guarantee Juve's progress to the last 16.
A statement on Juventus' official website read: "Gonzalo Higuain will not be on the plane to Spain after sustaining bruising to his left thigh in the final minutes of Saturday’s win over Pescara.
"The forward’s condition will be monitored closely over the coming days. Medhi Benatia, meanwhile, has not yet fully recovered from a knock to his right knee."
With Paulo Dybala still sidelined, Higuain's absence means Mario Mandzukic and 16-year-old Moise Kean are the only recognised strikers in the travelling party.
Kean would become the first player born this millennium to feature in the Champions League if he plays a part, having secured the same distinction in Serie A against Pescara.