Serbia axed injured centre-back Matija Nastasic and three other players as coach Mladen Krstajic named his final 23-man World Cup squad on Friday.
Schalke's Nastasic, a regular starter, failed to recover from a long-term knee injury while goalkeeper Aleksandar Jovanovic as well as midfielders Mijat Gacinovic and Nemanja Maksimovic were also left out.
The roster remains a blend of youth and experience with uncapped defenders Milan Rodic and Nikola Milenkovic as well as striker Luka Jovic joining winger Nemanja Radonjic, who made his debut in a friendly against South Korea in November.
Serbia start their World Cup campaign against Costa Rica on June 17, followed by games against Switzerland on June 22 and Brazil five days later.
They will play warm-up games in the Austrian city of Graz with Chile on Monday and Bolivia on June 9.
The full squad:
Goalkeepers: Vladimir Stojkovic (Partizan Belgrade), Predrag Rajkovic (Maccabi Tel Aviv), Marko Dmitrovic (Eibar)
Defenders: Aleksandar Kolarov (Roma), Branislav Ivanovic (Zenit St Petersburg), Dusko Tosic (Guangzhou R&F), Antonio Rukavina (Villarreal), Milos Veljkovic (Werder Bremen), Milan Rodic (Red Star Belgrade), Uros Spajic (Krasnodar), Nikola Milenkovic (Fiorentina)
Midfielders: Nemanja Matic (Manchester United), Luka Milivojevic (Crystal Palace), Sergej Milinkovic-Savic (Lazio), Marko Grujic (Liverpool), Adem Ljajic (Torino), Dusan Tadic (Southampton), Filip Kostic (Hamburg), Andrija Zivkovic (Benfica), Nemanja Radonjic (Red Star Belgrade)
Forwards: Aleksandar Mitrovic (Newcastle), Aleksandar Prijovic (PAOK Salonika), Luka Jovic (Benfica)