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Foreign Minister Penny Wong said 1500 Australians in Iran and 1200 Australians in Israel were seeking help to leave.
That number has more than tripled since Monday.
The government said earlier this week it was working on a range of plans to safely evacuate people from both countries, as Foreign Minister Penny Wong urged Israel and Iran to de-escalate the conflict.
Anybody wishing to flee either country is urged to register their interest on the Smartraveller website.
The current advice for Australians in Iran and Israel is to take shelter and listen to emergency warnings.
The ongoing conflict has also prompted Smartraveller to change its advice for Qatar on the Arabian peninsula.
“We now advise exercise a high degree of caution due to ongoing hostilities in the region,” the updated advice read.
“Conflicts in the Middle East could lead to an increase in tensions elsewhere in the region and may result in airspace closures, flight cancellations and other travel disruptions.
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“Demonstrations and protest activity may also occur, and local security situations could deteriorate with little notice. Avoid all demonstrations and protests.”
The warning demonstrates how far the impact of the regional conflict is already spreading.