The decision comes after the men's tennis world number one was drawn against fellow Serb Miomir Kecmanovic in the Australian Open, which is due to begin on Monday.
Novak Djokovic's visa has been cancelled by Australia's immigration minister, casting further doubt over his participation in the first grand slam of the year.
He first had his visa cancelled on arrival in Melbourne last week when his COVID-19 vaccination exemption was questioned.
But he won a court appeal against the cancellation that allowed him to remain in the country.
At the time, the Australian government said it would continue to look at whether he could stay, which was a decision that was entirely at the discretion of the country's immigration minister Alex Hawke.
"Today I exercised my power under section 133C(3) of the Migration Act to cancel the visa held by Mr Novak Djokovic on health and good order grounds, on the basis that it was in the public interest to do so," he said in a statement.
"In making this decision, I carefully considered information provided to me by the Department of Home Affairs, the Australian Border Force, and Mr Djokovic.
"The Morrison government is firmly committed to protecting Australia's borders, particularly in relation to the COVID-19 pandemic," Mr Hawke added.
Under the section of the Migration Act used by the minister to enforce his decision, Djokovic would not be able to secure a visa to come to Australia for three years, except in compelling circumstances that affect the country's interest.
It comes after the men's tennis world number one was drawn against fellow Serb Miomir Kecmanovic in the Australian Open, which is due to begin on Monday.
Djokovic could still file a legal challenge but if not his hopes of competing will come to an end.
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