London Irish centre Jonathan Joseph will almost certainly miss England's Test against Fiji after failing a fitness test on an ankle.
Joseph, who has been struggling with the injury since last week, could not train with the squad on Monday and England coach Stuart Lancaster does not expect him to be involved on Tuesday either.
The England players will have Wednesday off and Lancaster will name his team on Thursday which means Joseph is running out of time to prove his fitness.
Joseph's expected absence means that Brad Barritt and Manu Tuilagi are the favourite to start in midfield for England.
Lancaster's list of absentees includes the suspended Saracens wing Chris Ashton plus the injured quartet of Alex Corbisiero, Courtney Lawes, Dylan Hartley and Ben Foden.
Harlequins centre Jordan Turner-Hall is struggling with a knee injury, which is set to keep him on the sidelines for at least a fortnight.
Joseph, Turner-Hall were joined on the sidelines by Wasps lock Tom Palmer, who has a calf problem.
Gloucester duo Freddie Burns and Billy Twelvetrees and Wasps second-rower Joe Launchbury have joined the squad as cover for the injured trio but none of them are likely to feature against Fiji.
"Tom didn't train last week with a tight calf," Lancaster said.
"I just want to make sure I have all the bases covered. We were also keen to get Joe in, particularly in light of his recent form.
"It is an opportunity to bring Freddie into the equation because we recognise the form that he is in and we want to give him the opportunity to present himself up against Toby Flood and Owen Farrell.
"He can get a feel for what we are doing and get used to the game-plan because the reality is it only takes one injury (for him to be in the squad)."