Real Madrid superstar Cristiano Ronaldo was reportedly the target of homophobic slurs during Saturday's El Clasico at the Camp Nou.
Chants aimed at the Portuguese could be heard from some home fans during the minute's silence held for Barcelona legend Johan Cruyff before the match got under way.
According to Marca, the Spanish Observatory against lesbian, gay, bisexual and transgender phobia has now filed a complaint with the State Commission against Violence in Sport after it was revealed that Barcelona fans sang homophobic songs directed at Ronaldo during the weekend's derby
"These unfortunate and shameful acts are punishable, according to the Sports Act (2007)," Francisco Ramirez, director of the Observatory, said in a statement.
"Players and referees are repeatedly insulted with homophobic abuse. Exemplary punishment, to end this, is needed."
Ramirez also claimed that Ronaldo has been the subject of questions over his sexual orientation by journalists, players and fans of rival teams in order to humiliate him.
"It is necessary to clarify that homophobia does not necessarily mean that people who suffer are homosexual, but only that other people believe it or use it to insult, harass and humiliate others," he added.
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