The Spanish players were left fuming over the performance of official Vlad Iordachescu after the 18-10 defeat in Brussels allowed Romania to qualify for the World Cup at Spain's expense.
Ugly confrontations broke out at the final whistle as members of the Spanish coaching staff tried to restrain their players.
Several irate supporters also needed to be escorted off the pitch by competition officials.
“I am deeply saddened and disappointed by the attitude of the Spanish players, representatives and fans,” the Romanian federation chief, Alin Petrache, told Reuters on Wednesday.
“There is no room for hooliganism, misconduct and denigration in rugby."
The Spanish Rugby Federation lodged a formal complaint to World Rugby and Rugby Europe on Tuesday, demanding that the game against Belgium should be replayed due to the performance of the referee.
Spain's rugby federation condemned the reaction of "some of the players who could not contain their feelings".
Spain had needed to win to qualify automatically for their first World Cup since 1999, but now face a series of playoff games to reach the tournament in Japan next year.
Petrache, however, feels Spain are not ready to compete at the elite level.
“Letting one's emotions get the best of them is a sign of immaturity, that proves that the Spanish team is not ready to face responsibly the challenges an elite competition involves," he said.
“There is no room for this kind of violent behaviour in sport and, more importantly, in one of the few sports that still prides itself on its values and defends them.”
(Additional reporting by Joseph Cassinelli in Madrid; Editing by Toby Davis)
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