Victoria's coronavirus respite has been brief, with eight deaths taking the state toll to 737 and the national figure to 824.
Sept. 15 was the first day Victoria had been fatality-free in its second wave since July 13.
But there was more good news, with Melbourne's crucial 14-day average of new cases dropping below 50.
It is now 49.6, with regional Victoria also falling again to 3.5.
The 14-day rolling average is a key statistic in the state government's roadmap out of coronavirus restrictions.
Victoria had 42 new cases on Sept. 16, the same as Sept. 15.
Regional Victoria's restrictions will ease at midnight, but Melbourne will stay in its stage-four lockdown.
Authorities are ramping up roadblocks, creating the so-called "ring of steel" around the city to ensure city residents do not try to take advantage of the eased regional measures.
Premier Daniel Andrews has warned motorists travelling out of the city to expect longer wait times as police tighten checkpoints.
"I'm sorry to say it will mean that there will be significant queues, there will be travel issues," he told reporters on Sept. 15.
Melbourne's lockdown rules remain unchanged and people cannot travel out of the city without specific reasons.
The next step for regional Victoria means pubs, cafes and restaurants will be able to serve people outside with strict density quotas, while outdoor gathering limits will be upped to 10.
Regional Victorians will also be able to leave their homes without restriction and all shops can reopen.
The premier said Melburnians should be inspired by the rolling back of restrictions, rather than disheartened.
"I'd encourage people not to see it that way and instead see this as proof positive," he said.
Melbourne will move to its next step of reopening on Sept. 28 if the 14-day case average keeps falling to between 30-50.