All AFL and AFLW players will need to be fully vaccinated against COVID-19 by mid-February or they will be barred from playing and training.
The AFL has released its long-awaited vaccination policy, with clubs to ultimately determine action on staff who refuse the jab.
If players do not have a medical exemption, clubs will have options to transfer them to the inactive list, pay them no less than 25 per cent of their contracted salary, or agree to part ways.
The AFL's vaccination schedule will be rolled out across three stages, but all players will be required to have the jab if they want to continue their playing careers.
The Victorian government's vaccination requirement about a range of workers, which includes footballers, came into effect last Friday.
Seven of the eight AFLW teams have confirmed to AAP that their women's programs are compliant with the vaccine protocols and players have been cleared to train and play.
But the AFL's announcement on Thursday forces players in other states to be vaccinated.
Sydney Swans and GWS players will need to be fully vaccinated by December 17, while clubs in Queensland, Western Australia and South Australia will need to have both doses by February 18.
"The uptake of AFL industry people being vaccinated has so far been extremely encouraging and we are very pleased the overwhelming majority of our players, coaches, staff members across the AFL and the 18 clubs have at the very least received their first vaccination shot," AFL football general manager Andrew Dillon said
"Our policy delivers on our commitment to best protect our players, staff and the wider community, so we can once again unite as families, as friends, as work colleagues, as teammates, as supporters, as communities, as one."
Adelaide's Deni Varnhagen this week became the first known AFL or AFLW player to refuse the COVID-19 vaccination.
A registered nurse, who has worked in intensive care units, Varnhagen informed the Crows she is not yet willing to receive the jab.
Varnhagen, who played in the Crows' 2017 and 2019 premierships, posted a video on social media on Saturday showing people marching in Adelaide for "freedom of choice".
AFL Players Association boss Paul Marsh said it became clear unvaccinated players would be unable to be part of the competition.
"In our conversations with players about COVID-19 vaccinations over recent months, we have made it clear that the AFLPA believes in the importance of vaccinations," Marsh said.
"In saying this, our view is that vaccinations are an individual decision.
"It has, however, became apparent in recent weeks that unvaccinated players will not be able to fulfil their contractual obligations due to various state border restrictions.
"In working through this policy with the AFL, our focus was to agree positions that provide players with clarity, appropriate timelines for making personal vaccination decisions, an ability to review the policy should government directions change, and financially support those who make the hard decisions to step away from their playing careers."
Melbourne premiership hero Tom McDonald strongly came out against mandatory vaccinations for players, calling it "ethically wrong", but said he had already received both doses.