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A spike in sick leave among paramedics reduced the number of ambulance crews operating across Victoria overnight, according to the union.
At least 50 crews were taken off state roads, with crews dropped from metropolitan and rural areas.
The Victorian Ambulance Union said members reported crews had to drive more than 60km to reach Code 1 cases, which are considered high priority and time critical incidents.
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About 20 crews were dropped from rural areas including Hamilton, Barwon Heads, Norlane, Warrnambool, Swan Hill, Heathcote, Bendigo and Mildura.
Meanwhile, at least 30 were dropped in Melbourne, including Bayswater, Brighton, Sunshine, Doncaster, Rowville, Mordialloc, Hartwell, North Melbourne and Oak Park.
"The members rarely get breaks, almost never finish on time and they are exhausted and burnt out. So, this is the reality we are facing," Victorian Ambulance Union secretary Danny Hill said.
"We have been warning about the need to better support the workforce, so we have a healthy and well supported workforce.
"If you don't support the workforce, then the response to the community suffers".
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There are normally 120 ambulances working night shifts each night. Last night only 90 were in action, according to the union.
It comes just weeks after Ambulance Victoria chief executive Jane Miller resigned amid mounting pressure from ongoing industrial disputes and increasing work demands.
Ambulance Victoria has been approached for comment.
