Joe Allen has been ruled out of Wales' Euro 2016 qualifier against Israel with a hamstring injury.
The Liverpool midfielder was suspended for the 3 September trip to Cyprus but will also miss the game with Israel three days later in Cardiff.
Defender Adam Matthews is also out after being injured in Sunderland's 6-3 League Cup win against Exeter.
Real Madrid forward Gareth Bale is included having declared his fitness after a toe injury worry.
"When it initially happened [Bale's injury] you're holding your breath a bit, of course," admitted Wales manager Chris Coleman.
"But the scan and the feedback was positive thankfully so I don't think there's going to be a problem.
"There have been constant conversations with both medical teams - there's a very good relationship there - but... it wasn't as bad as first thought."
James Collins, Ben Davies and David Edwards return after missing June's 1-0 win over Belgium through injury.
But Nottingham Forest midfielder David Vaughan has withdrawn for family reasons, while fellow midfielder Joe Ledley has not been playing regularly this season for Crystal Palace.
Allen's Liverpool team-mate and fellow midfielder Jordan Williams, who is currently on loan at League One side Swindon Town, is included and could win his first cap.
"Obviously Joe Allen was suspended for the first game... but we were optimistic for the second one up until recently," said Coleman.
"So it's disappointing because we haven't had Emyr Hughes [the Wigan midfielder who has a long-term ankle problem] for a while through injury, who can play in that holding position like Joe.
"We've lost David Vaughan also, so with natural holding players it's been a bit tough."
Wayne Hennessey's inclusion should see the Crystal Palace goalkeeper win his 50th international cap against Cyprus in Nicosia.
Wales have a three-point lead at the top of Euro 2016 qualifying Group B with four games remaining.
Wins in both games against Cyprus at the GSP Stadium in Nicosia and Israel at Cardiff City Stadium could be enough to seal Wales' place at a major finals for the first time since 1958.
"This period of my career is my biggest. I know what the dangers are," Coleman said.
"You can feel it around the country: the belief is there, the excitement is there, the anxiety. The pressure is massive but this is what we've always wanted."
The top two sides from each group qualify automatically for the Euro 2016 finals in France, with Wales holding a five-point lead over third-placed Israel going into September's games.
Israel will be without star striker Omer Damari for their game on 6 September, as the 26-year-old Red Bull Salzburg player has been ruled out with a broken foot.
Damari has five goals in this campaign, making him joint top scorer in the section with Wales' Bale and Bosnia-Hercegovina striker Edin Dzeko.
Goalkeepers: Wayne Hennessey (Crystal Palace), Danny Ward (Liverpool, on loan with Aberdeen), Owain Fon Williams (Inverness Caledonian Thistle); Defenders: Ashley Williams (Swansea City), James Chester (West Bromwich Albion), James Collins (West Ham United), Ben Davies (Tottenham Hotspur), Chris Gunter (Reading), Neil Taylor (Swansea City), Adam Henley (Blackburn Rovers), Ashley Richards (Fulham); Midfielders: Andy King (Leicester City), Joe Ledley (Crystal Palace), Aaron Ramsey (Arsenal), Jordan Williams (Liverpool, on loan with Swindon Town), David Edwards (Wolverhampton Wanderers), Shaun MacDonald (AFC Bournemouth); Forwards: Gareth Bale (Real Madrid), Hal Robson-Kanu (Reading), Sam Vokes (Burnley), Simon Church (MK Dons), David Cotterill (Birmingham City), Tom Lawrence (Leicester City, on loan with Blackburn Rovers).