The annual vessel exclusion zone comes into effect from Monday, June 1, to protect southern right whales involved in the annual migration.
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Boat operators are being reminded to steer clear of Warrnambool's Logans Beach to protect whale mothers and their calves sheltering in the nursery area.
All powered vessels, including boats and jet skis, and aircraft, including drones and helicopters, cannot enter the exclusion zone.
The fine for any vessel entering the exclusion zone is $3170.
Conservation Regulator program manager Ash Burns said it was critical people adhere to the regulations.
"Logans Beach is the only established nursery in south-east Australia where female southern right whales raise their calves, so it's vital we give them space to rest and nurse their young," he said.
"The exclusion zone is in place for everyone's safety, as female southern right whales are known to react to watercraft to protect their calves."
Outside of the Logans Beach exclusion zone, recreational boaters must remain 200 metres from whales, while high impact vessels, such as jet skis, have to be 300 metres away from whales.
Swimmers must not approach within 50 metres of a whale and aircraft are not permitted within 500 vertical metres if the aircraft is operating within a 500-metre radius of a whale.
The whale viewing platform is open with social distancing and group limits applying.
The south-eastern Australian southern right whale population is estimated to be less than 300 individuals and is listed as critically endangered in Victoria.
Anyone who observes vessels within the exclusion zone or breaching minimum distances is urged report to Crime Stoppers on 1800 333 000.
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