Wolves manager Mick McCarthy and his players slammed Wigan defender Antolin Alcaraz after television replays appeared to show him spitting at right-back Richard Stearman.
The incident occurred in the 83rd minute of Wolves' crucial 3-1 home victory and sparked a brawl involving several players from each team.
Referee Lee Probert took no action but Stearman claimed Alcaraz had spat in his direction after it was initially thought it was aimed at Christophe Berra.
Stearman was reluctant to talk about the incident but said: "It wasn't Berra, I'll give you that, but I don't think it's right for me to talk about it at the moment."
Asked if he had been spat at, Stearman added: "Yes, but I'm not going to speak about it."
McCarthy said: "Spitting is taboo if that's what happened. I only had it happen to me once and the player didn't come too close to me for a long time after that.
"If it has been shown, that is unacceptable behaviour and anyone here who was spat at would act accordingly.
"If that has happened, he shouldn't be doing it."
David Edwards, who scored Wolves' second goal, said: "That is out of order. That shouldn't be brought into the game and hopefully he should be pulled up on that.
"I'm sure with the television footage as well, some sort of action should be taken."
Wigan boss Roberto Martinez would only say: "I haven't seen (the replays) yet. If that is the case, I'll look into it and I'll find out."
The incident overshadowed Wolves first win in nine games as goals from Jamie O'Hara, Edwards and Stephen Ward clinched the three points.
Alcaraz is now facing a three-match ban after being charged by the Football Association with spitting at an opponent.
The FA said in a statement on Monday afternoon: "Wigan Athletic's Antolin Alcaraz has been charged by the FA for spitting at an opponent.
"The incident, which was not seen by the match officials but caught on video, relates to his side's game against Wolverhampton Wanderers.
"The player has until 6pm on November 8, 2011 to respond to the charge. If the player admits the charge he may accept an automatic three-match suspension."
