Rovers boss Jon Dahl Tomasson says Arnor Sigurdsson and Jack Vale will be out of action “for a good while”.
Iceland international Sigurdsson, who joined from CSKA Moscow, is yet to make his first appearance after picking up a groin injury in training.
Meanwhile, Vale injured his hamstring during the recent training camp in Austria.
The pair are now facing a spell on the sidelines as Rovers prepare for the upcoming Championship campaign.
Adam Wharton has recovered from a muscle injury but missed the goalless draw at Fleetwood through illness.
Niall Ennis is also set to step up his training schedule early next week following his free transfer from Plymouth.
“Arnor and Jack will not be back for a good while,” said Tomasson. “Adam is back but he got sick, so I am not sure if he will be playing (against Barnsley).
“I am disappointed for Adam after he was not involved in Austria. He is fit but now he is sick.
“(Niall) needs to be fit first. If everything goes to plan, he will start getting involved on Monday in a bit of the training, building up like that.”
Tomasson also confirmed Callum Brittain is fine despite needing treatment during the first half at the Highbury Stadium.
Rovers had several good opportunities against Scott Brown’s side but failed to break the deadlock.
“We had a couple of times where the quality of the pass in the box maybe was not the best,” the manager explained.
“But we still created plenty of good opportunities to score goals. The second half was okay, the first half was not good enough.”
He added: “A clean sheet is always good but at the moment, it is about working on the fitness and getting to see the principles of what I want to see. We saw that, for sure, in the second half.”
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