Jonathan Joseph has effectively been ruled out of England's opening QBE autumn international against Fiji after failing a fitness test on an ankle injury.
The London Irish centre was unable to train again with the squad on Monday afternoon and head coach Stuart Lancaster did not expect him to be involved on Tuesday either.
With the England players having Wednesday off and Lancaster scheduled to name his team on Thursday, Joseph is out of time to prove his fitness to face the Fijians.
Joseph's absence increases the likelihood of England starting against Fiji with a centre pairing of Brad Barritt and Manu Tuilagi, with Alex Goode at full-back and operating as a second play-maker in the Red Rose back division.
Lancaster is having to manage a lengthening list of absentees for Saturday's Twickenham clash that also 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 expected to be sidelined for at least a fortnight by a knee injury, which opened the way for Gloucester fly-half Freddie Burns to join England's training squad.
Burns' Kingholm team-mate Billy Twelvetrees has also joined the squad as Joseph's injury replacement and Wasps lock Joe Launchbury is providing precautionary cover for Tom Palmer.
None of those three late call-ups are likely to feature against Fiji - and Palmer has returned to training after a calf problem - but Lancaster wanted to reward their Aviva Premiership form and give them a taste of the England environment.
Joe Marler has shaken a hamstring problem which stopped him training last week and number eight Ben Morgan is available for selection, despite requiring 15 stitches to a head wound in Gloucester's Aviva Premiership defeat to Harlequins.
The further good news for Lancaster is that Corbisiero is due to play for London Irish this weekend, with a view to him being in consideration to face Australia on November 17.