Thirty-nine vegan champions who choked central Melbourne with their calls to end animal cruelty will face a magistrate on obstruction charges.
In a surprise for police during Monday's morning peak commute, more than 100 activists blocked the jam-packed intersection outside Flinders Street Station.
Three rental vans were used in the blockade as activists chanted for "animal liberation", with some sitting on tram tracks, linking arms in a circle to halt police attempts to break up the protest.
A number of people also chained themselves to the vehicles that were draped in black and emblazoned with the web address of a vegan film documentary.
The activists didn't just face opposition from police, with a number of hecklers slamming the protest, including a man wearing a clown mask who yelled "eat more meat".
Protesters also gathered at abattoirs in Corio, Pakenham, Laverton North and Bacchus Marsh, as part of a national series of rallies.
Two 17-year-olds, a 15-year-old and 36 adults were charged with offences relating to obstructing a roadway and resisting police.
They have been released on bail and will face court at a later date.
Once the two-hour Melbourne intersection protest was disbanded, some activists moved to block the entrance to the Melbourne Aquarium, where another person was arrested and later charged.
Protest organiser and documentary director, Chris Delforce, said the protest marked one year since the release of his film, Dominion.
"The industry is telling people these animals are being killed ethically, that they are being killed humanely. The reality is ... it's the furthest thing from humane," Mr Delforce told AAP.
Victoria Police Superintendent David Clayton criticised the protesters for not consulting police before the event.
"We respect the right of people to protest peacefully, but we will not tolerate anti-social behaviour and people committing offences," he told reporters.
The demonstration caused some trams to alter course while pedestrians, including children on their first day of Victorian school holidays, were unable to cross the Swanston-Flinders Street intersection.
"State and territory governments should ensure the full force of the law is brought against these green-collared criminals," Prime Minister Scott Morrison said.
Monday's protest comes after the Gippy Goat Cafe in West Gippsland announced on Sunday it had closed, blaming "constant harassment, vile statements and threats from the abusive vegan activists".
Victorian Nationals leader Peter Walsh said it was "gutting" to see farmers targeted.
"If current Victorian laws aren't protecting our farmers and the integrity of Victoria's biosecurity they must be strengthened," he said.
Among their demands, protesters want state and federal agricultural ministers to "acknowledge cruelty in the process of killing animals for food, clothing and entertainment" and to add warning labels on animal products.
Source: Aussie Farms
Australian Associated Press