Unvaccinated staff, including 25 paramedics, were notified this week their employment with Ambulance Victoria would end.

They had been asked to take leave to consider their future positions, after failing to comply with the directive for a first dose by October 15. Second doses are required by next week.

Six employees have been granted exemptions by Ambulance Victoria, where 99 per cent of the workforce has been vaccinated.

But more than two dozen others have filed a case in the Fair Work Commission arguing they were denied procedural fairness over the policy, claiming their termination goes beyond the state's directive.

In a statement, lawyers for the staffers say at least 29 have been either stood down or "coerced into compliance" to retain their jobs.

"We were called heroes last year, this year we're accused of serious misconduct and our employment is terminated," paramedic Jessica Davis said.

Ms Davis is one of 23 employees already sacked, while the remainder have meetings scheduled for later this week, she told a Fair Work Commission hearing before deputy president Amanda Mansini on Tuesday.

Their lawyer, Benedict Coyne, has requested an interim order preventing changes to their employment ahead of conciliation or, if that fails, arbitration.

Ambulance Victoria's lawyer Jack Tracey said he had a "lack of optimism" about whether conciliation would achieve anything, given the nature of the matter.

Ms Davis has accused Ambulance Victoria of a significant overstep, saying the chief health officer's directives on vaccine mandates do not require employees who fail to comply to be terminated.

"Staff that have been working from home for the last 18 months are now fired because AV state they must be able to enter an AV workplace premises," she said.

"This goes beyond the CHO's directions."

Ms Davis claimed staff were degraded or mocked when they sought support for their position.

Their lawyers say they are not anti-vaccination and are medically informed.

"Several members of the group felt coerced into having a first dose and are prepared to get a second dose but want to wait until a proven safe vaccine becomes available such as Novavax and COVAX," the statements said.

The Therapeutic Goods Administration has provisionally approved four vaccines - AstraZeneca, Pfizer, Moderna and Johnson & Johnson - as safe and effective for use in Australia.

An Ambulance Victoria spokeswoman said unvaccinated staff had been asked to take leave to consider their positions.

"Having carefully considered all individual circumstances, unvaccinated staff have been notified that their employment with Ambulance Victoria will end, including 25 paramedics," she said.

Ambulance operations are not expected to be affected.

A further hearing in the FWC will take place on Wednesday.

Australian Associated Press