Wales scrum-half Rhys Webb has been ruled out of the World Cup with a foot injury, the Welsh Rugby Union has announced.
Webb, last season's Welsh player of the year, was injured during Wales' 23-19 World Cup warm-up victory over Italy in Cardiff on Saturday.
"Scrum-half Rhys Webb has been ruled out of @rugbyworldcup with foot injury. Further examination on foot and knee to come," the WRU said on its official Twitter account.
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The news comes on the back of Monday's announcement that British and Irish Lions full-back Leigh Halfpenny would miss the World Cup after suffering a knee ligament injury against Italy.
Wales head coach Warren Gatland has called Mike Phillips and Eli Walker into his World Cup squad to cover the two injuries.
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Lions scrum-half Phillips has won 94 caps for Wales and was a surprise omission when Gatland named his 31-man tournament squad, while Ospreys wing Walker made his international debut against Ireland last month.
"It is extremely disappointing for both Leigh and Rhys," Gatland said.
"They have worked exceptionally hard to establish themselves on the world stage and to prepare for the tournament and we wish them all the best with their recovery.
"After announcing the squad we spoke about the need for players to remain ready for action and is a great opportunity for Mike and Eli.
"It's great that we can call on players of this calibre. To bring Mike's experience into the squad is a bonus along with Eli's exciting talent."
Webb's absence from the World Cup makes it a desperate couple of days for Wales following the loss of Halfpenny.
Toulon points machine Halfpenny stands third on Wales' all-time scoring chart behind Neil Jenkins and Stephen Jones with 508 points in 62 Tests and his absence from the tournament is a huge blow to Gatland's side.
Wales legend Shane Williams has questioned Gatland's decision to field Halfpenny against Italy given the 26-year-old's importance to the national team.
"It's just horrendous for Wales," said Williams at the launch of ITV's World Cup coverage.
"Rhys Webb has been the form player for Wales over the last two seasons. To lose Halfpenny as well, it's just a massive blow for Wales and it couldn't have come at a worse time.
"You kind of thought 'do we need this guy on the field? Let's wrap him up in cotton wool and roll him out when we really need him'.
"It is a massive blow and Wales is in mourning already. Especially with Halfpenny, he was strapped up on his leg anyway.
But Wales centre Jamie Roberts said: "Warm-up games are important, for the lads to get up to match fitness.
"These injuries can happen in training, and can happen in games. Hindsight's a wonderful thing. If we beat Italy by 60 points at the weekend with a clean bill of health it's a great exercise, so what can you say?
"It's very easy to make comments like (Williams') after.
"But it's a blow, there's no doubting that."
Webb issued a statement after he had been ruled out of the World Cup, expressing his disappointment at the news but also insisting that Wales will do well in the tournament.
Wales start their World Cup campaign against Uruguay in Cardiff on September 20 and also line up against tournament hosts England, Australia and Fiji in Pool A.