The Biancocelesti chief hit out at supporters who had urged the Stadio Olimpico side to let Juventus win, preventing Roma from catching up in the title race.
Defeat to the Bianconeri would deny fierce local rival Roma the chance to close the gap on the Turin outfit at the top of Serie A, prompting Biancocelesti fans to make the case for intentional defeat on radio stations.
But Lotito claims the severity of the rivalry is having a negative impact on the matchday experience at the Stadio Olimpico and threatened to punish those who called for the loss.
"What the fans do will not remain unpunished," he told Tuttosport. "People must understand that football is to be enjoyed, not treated with hatred, bitterness or violence. You must go to the stadium to enjoy a wonderful day and at the end the winners and losers emerge in a positive atmosphere, regardless of the result. That’s what happens in English stadiums.
"Juventus also have a section of their fans who create problems. I have always said we need football to teach and set the moral tone. Once anything was allowed, now there is zero tolerance. It’s still not enough, as we must all understand that in football there are opponents, not enemies."
Lazio hosts Antonio Conte's men on Saturday.