Police have arrested a total of nine people since protests at the McDonald's proposed site in Tecoma, in the Dandenong's, began.
The most recent arrest was early this morning, when a single protester on the roof of the building, was removed from the site.
She has since been charged with trespass.
"She was passively resistant," Acting Superintendent Paul Rosenblum said.
The woman who had been camped on the roof, was taken down with the use of a cherry picker.
About 50 police officers were deployed to the area early on Tuesday morning, to make way for construction of the McDonald's development.
Acting Sup. Rosenblum has defended the use of police resources used at Tuesday's operation.
"We assess and plan every operation on a needs basis," he said.
"Heavy machinery was brought into the site, and previous indications were for the potential for people to put themselves in harms way.
"Clearly our role is to ensure no-one is breaking the law."
He says in total there have been nine arrests.
"Earlier on, there were 8 people arrested, and charged in relation to matters pertaining to the construction site at Tecoma."
One protester says they have been maintaining a peaceful demonstrators.
"We'll maintain the blockade and continue to maintain our protest," he said.
Asbestos will be removed from the building, to pave way for construction to begin.
"They're going to have to pick us off the road and remove us," said the protester.
"McDonald's must be wondering about the PR that's generating from this, because it's damaging their reputation, it is hurting them."
The protesters say their petition now has almost 80,000 signatures.
Another protester says the outlet will produce a lot of litter in the area.
"It's just down the road from a national park, there's a kindergarten and primary school on the other side of the road, and it's a very congested road and will probably get worse,"
He says their list opposing the McDonald's development, goes on.
"A lot of us a working, we're not unemployed hippies," he said.
One mother says she came to the protest, because of her children's interest in the campaign.
"I've been here quite a few times, but today I thought I'd give the little ones a go," she said.
A McDonald's spokeswoman says the company has always respected people's right to express their opinions, but not when it puts themselves or others at risk.