Italy coach Cesare Prandelli dumped Daniele De Rossi from his squad after the Roma midfielder's punch on Inter's Mauro Icardi.
Prandelli announced his squad to face Spain on Sunday and De Rossi was the most notable omission, after he was caught on camera punching Icardi in the face during Roma's clash with Inter on Saturday.
Pablo Osvaldo and Mario Balotelli have previously been left out by Prandelli for misdemeanours at club level and, while De Rossi avoided sanction during Saturday's match, his punch has been discussed ad nauseam since in the Italian media.
It was better news for De Rossi's club team-mate Mattia Destro, who has hit the back of the net six times in 11 games in the Serie A since returning from a knee injury.
Destro was recalled by Prandelli, as were Genoa goalkeeper Mattia Perin and Milan defender Mattia De Sciglio.
Italy will face Spain in Madrid on Wednesday with two uncapped players included in the squad - Parma defender Gabriel Paletta and Torino forward Ciro Immobile.
Milan striker Mario Balotelli (shoulder) remains sidelined.
Squad in full:
Goalkeepers: Gianluigi Buffon (Juventus), Mattia Perin (Genoa), Salvatore Sirigu (Paris St Germain)
Defenders: Ignazio Abate (Milan), Davide Astori (Cagliari), Andrea Barzagli (Juventus), Leonardo Bonucci (Juventus), Giorgio Chiellini (Juventus), Domenico Criscito (Zenit St Petersburg), Mattia De Sciglio (AC Milan), Christian Maggio (Napoli), Gabriel Paletta (Parma)
Midfielders: Antonio Candreva (Lazio), Emanuele Giaccherini (Sunderland), Claudio Marchisio (Juventus), Riccardo Montolivo (AC Milan), Thiago Motta (Paris St Germain), Marco Parolo (Parma), Andrea Pirlo (Juventus), Marco Verratti (Paris St Germain)
Forwards: Alessio Cerci (Torino), Mattia Destro (Roma), Alberto Gilardino (Genoa), Ciro Immobile (Torino), Gonzalo Insigne (Napoli), Pablo Osvaldo (Juventus)