Midfielder Daniele De Rossi was left out of Italy's squad for the friendly against Spain, one day after television cameras caught him punching an opponent in an off-the-ball incident during a Serie A match.
The Roma midfielder was seen to land a punch on Inter forward Mauro Icardi's jaw during their goalless draw yesterday although the incident was missed by match officials.
It was the latest in a long line of misdemeanours by De Rossi, who was also sent off for a wild two-footed challenge during Roma's game against Juventus in January.
Italy coach Cesare Prandelli has frequently dropped players, including De Rossi twice before, for bad behaviour in club matches under his strict code of ethics.
Pablo Osvaldo and Mario Balotelli have also fallen foul of Prandelli's rules in the past.
Balotelli was omitted this time because of a shoulder injury while uncapped Parma defender Gabriel Paletta and Torino forward Ciro Immobile were given their first call-ups.
Forward Mattia Destro, who has scored six goals for Roma since returning to action in December after a lengthy injury saga, was recalled along with Genoa goalkeeper Mattia Perin and Milan defender Mattia De Sciglio.
Italy, beaten 4-0 by Spain in the Euro 2012 final, visit the world and European champions in Madrid on Wednesday.
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) (Reporting by Brian Homewood; editing by Toby Davis)