Striker Josh Kennedy and playmaker Tom Rogic have been cut from the Socceroos' final World Cup squad because of injury.
Kennedy scored the goal against Iraq last year that secured Australia's spot in Brazil but won't play a part in what would have been his third consecutive World Cup campaign.
In another surprise omission, defender Luke Wilkshire was left out.
With 79 caps, Wilkshire was the most experienced player in the squad and, aside from Mathew Spiranovic, was the only defender with more than 10 international appearances.
With Ivan Franjic still recovering from a knock to his knee, 32-year-old Wilkshire was a viable option at right back for what would have been his third World Cup.
Newcastle Jets goalkeeper Mark Birighitti was the fourth player to miss out on coach Ange Postecoglou's 23-man squad, with Mat Ryan, Mitch Langerak and Eugene Galekovic picked ahead of him.
Kennedy was due to play in Australia's warm-up match against Brazilian division two side Parana Clube on Monday (Tuesday AEST) but was sidelined after his ongoing back injury flared up in training the previous day.
Rogic did play but, with persistent groin concerns and a lack of game time, he was forced off the field after a lacklustre first-half showing.
Postecoglou met with his coaching and medical staff on Monday night (Tuesday AEST) to decide the cuts.
Postecoglou said the decisions were tough but ultimately came down balancing the squad and ensuring those selected were fit enough to stand up to rigours of facing Chile, the Netherlands and Spain.
"Every step of the selection process has been difficult because the players have worked very hard since they came into camp but we have had to weigh up form, fitness and squad balance," he said.
"We've selected a squad that can do Australia proud and they will walk out onto the pitch for the World Cup in the green and gold shirt and display the characteristics that define our country.
"Our stated aim is to make our country proud, show the world what Australians are made of and test ourselves against the very best football players in the world and this group of players is capable of doing that."
Australia have a crucial final warm-up match against Croatia on Friday (Saturday AEST) before opening their World Cup campaign against Chile on June 13.
Group matches against the Netherlands (June 18) and Spain (June 23) follow.
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