Blackburn Rovers will assess John Fleck's early injury against Preston North End whilst John Buckley will play for the Under-21s on Monday.
Fleck was handed his Rovers debut at Deepdale but was forced to withdraw around the 15-minute mark. The Scotsman went into a full-blooded challenge and came out ok before appearing to hurt himself with the next action.
Eustace wasn't aware of what the specific problem was but admitted it was a blow to lose the experienced midfielder at that stage in the match.
"It's a massive blow to lose him," Eustace said.
"He has come in to help us with his experience. I thought it was the perfect game for him to come in and start.
"I think he took a kick to his calf and we will assess it. He started the game very well. I don't know what the injury is but we will assess that in the morning and take it from there."
Harry Leonard missed out on a place in the matchday squad due to a small injury whilst Joe Rankin-Costello was ruled out before the weekend.
Buckley was amongst the travelling Rovers party but wasn't selected. Eustace instead wants the midfield to play against Derby County for the U21s on Monday evening.
The 24-year-old has been out for two and a half months after shoulder surgery and hasn't played much football for either Rovers or Sheffield Wednesday in the last six months.
"Harry has got a little knock, unfortunately," Eustace said.
"Bucko hasn't had much game time. He was part of the (travelling) squad today.
"He'll be playing on Monday and I look forward to watching him play. He's a player I admire a lot, he's done very well for the football club over the years.
"I am looking forward to having him back in the squad as well."
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