About 100 Victoria Police staff, including Chief Commissioner Shane Patton, were among the more than 15,000 marchers who took part in the annual event in St Kilda on Sunday.
Police were confronted by up to 50 protesters as they marched, with the force accusing the group of surrounding its members on three sides, hurling abuse and throwing paint bombs.
People march along Fitzroy Street during the Midsumma Pride March in St Kilda. (Diego Fedele/AAP PHOTOS)
"Their conduct, I have nothing but contempt for it," Mr Patton said.
"Someone was in a pig's head ... their intent was clear."
Vision of the clash shows police and protesters pushing and shoving each other as chants of "cops kill queers" rang out.
A banner also read: "No pride in prisons. Vicpol + Serco queers hate you."
One protester, a 34-year-old man, was arrested for assaulting a spectator but released without charge.
One of the activist groups involved in the protest, the Civil Resistance Youth Movement, accused police of throwing protesters to the ground and punching one in the side of the head.
They also disputed Mr Patton's suggestion that paint bombs were thrown.
"There were only a few bottles of paint and a single coffee cup, and the only intended target of paint were protesters, for theatrical purposes," the group said.
The march is one of the signature events at the Midsumma Festival, which celebrates gender and sexual diversity.
But police participation in pride events is controversial within the LGBTQI community, given their origin out of the protest movement against police oppression of queer people.
Midsumma Festival chief executive Karen Bryant expressed deep disappointment with people from "both sides" for actively escalating the volatile situation.
"Our immediate focus is directed towards providing support for injured staff members, and we are still collating and viewing documentation of the incidents that occurred on the day," she said in a statement.
"Feeling safe, supported, and included is a fundamental human right, and it is disheartening that the events that occurred at the Pride March did not uphold these principles.
"Midsumma commends its staff for their efforts in managing and de-escalating a challenging and volatile situation displayed by both sides of the incident."
Mr Patton said he was pleased with the restraint shown by the public order response team under the circumstances.
"We certainly didn't escalate it," he told ABC radio on Monday.
Police are also reviewing footage of the clash to determine if further action is required.
Victorian Police Minister Anthony Carbines said the actions of protesters should be condemned without equivocation.
"You don't get to bring your views to the table and discuss them or promote them when you're acting like a thug," he told reporters on Monday.
"Yesterday was just a concerted and premeditated attack on members of Victoria Police who were participating in a community event.
"Nobody accepts that. It's not tolerated and there are no excuses for it."
Mr Carbines, who was scheduled to meet with Mr Patton on Monday, indicated the government would likely discuss the issue directly with festival organisers.
"There are elements that are going to need to be addressed," he said.
"Because everyone deserves to be able to participate ... in a safe way and not to be threatened or assaulted. It doesn't matter who you are."
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