Tottenham full-back Danny Rose and Southampton goalkeeper Alex McCarthy have withdrawn from the England squad for this month’s Nations League matches with minor injuries.
The pair have returned to their clubs before Gareth Southgate’s squad fly out to face Croatia on Friday.
Both arrived at the camp with pre-existing injuries and will not be replaced in the 25-man squad.
Rose’s absence may mean a first senior start for Leicester’s Ben Chilwell.
Twenty-one-year-old Chilwell, who was called up to replace injured Manchester United defender Luke Shaw at the weekend, is the only specialist left-back in squad. He made his debut as a replacement for Rose during September’s friendly win over Switzerland.
Rose started two matches during the summer’s World Cup campaign – the group-stage and third-place play-off defeats by Belgium – as Southgate mostly opted for a three-man defence with Ashley Young as a wing-back on the left.
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Even after the withdrawal of the uncapped McCarthy, Southgate has three goalkeepers at his disposal with Fulham’s Marcus Bettinelli, Stoke’s Jack Butland and Everton’s Jordan Pickford still involved.
On Sunday, uncapped Brighton defender Lewis Dunk, 26, was drafted in after an injury to Burnley’s James Tarkowski.
England play Spain, who inflicted a 2-1 defeat in their Nations League opener last month, in Sevilla on Monday.