Middlesbrough forward Scott McDonald was dropped in a surprise move as Australia trimmed its World Cup roster from 31 players to 28.
Crystal Palace midfielder Nick Carle and Norway-based defender Jade North also were cut Tuesday.
McDonald, who played in Monday's 2-1 exhibition win over New Zealand, had been a prolific goalscorer in Scotland before moving to England, but has yet to score in 16 matches for Australia.
Belgian-based Nikita Rukavytsya and Josh Kennedy will be the backups to Galatasaray forward Harry Kewell.
Australia coach Pim Verbeek said McDonald struggled in the Socceroos' preferred lone striker system.
"Scotty is a typical two-striker player, and that's where he feels happy also," Verbeek said. "But we don't play with two strikers. We play with one striker up top and wingers, so Scotty doesn't fit in this system. He knows it, and he tried everything."
The core of Australia's squad is made up of veterans of the 2006 World Cup, headed by Kewell, clubmate Lucas Neill, Everton's Tim Cahill, Palermo's Mark Bresciano and Fulham goalkeeper Mark Schwarzer.
Verbeek said players who did not make his final 23-man roster, which must be submitted to FIFA on June 1, will be kept on in South Africa for the duration of the World Cup. Youngsters Tommy Oar and James Holland are unlikely to make the final 23 but have been chosen to gain experience.
Australia leaves on Wednesday for a pre-World Cup training camp in Johannesburg. The team will play exhibitions against Denmark on June 1 and the United States on June 5 ahead of its World Cup opener against Germany on June 13.
Goalkeepers: Adam Federici (Reading, England), Eugene Galekovic (Adelaide United). Brad Jones (Middlesbrough, England), Mark Schwarzer (Fulham, England)
Defenders: Michael Beauchamp (Melbourne Heart), David Carney (Twente, Netherlands), Scott Chipperfield (Basel, Switzerland), Shane Lowry (Aston Villam England), Mark Milligan (JEF United, Japan), Craig Moore (no club), Lucas Neill (Galatasaray, Turkey), Luke Wilkshire (Dynamo Moscow, Russia), Rhys Williams (Middlesbrough, England)
Midfielders: Mark Bresciano (Palermo, Italy), Tim Cahill (Everton, England), Jason Culina (Gold Coast), Brett Emerton (Blackburn, England), Richard Garcia (Hull, England), Vince Grella (Blackburn, England), James Holland (Alkmaar, Netherlands), Brett Holman (Alkmaar, Netherlands), Mile Jedinak (Antalyaspor, Turkey), Tommy Oar (Utrecht, Netherlands), Carl Valeri (Sassuolo, Italy), Dario Vidosic (Nuremberg, Germany)
Forwards: Josh Kennedy (Nagoya Grampus, Japan), Harry Kewell (Galatasaray, Turkey), Nikita Rukavytsya (Twente, Netherlands)