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Team officials have confirmed Atkinson had not recovered from a knock he suffered in the team's devastating defeat at the hands of France on Thursday.
Atkinson's absence is a big loss for the Aussie side, who face an enormous challenge against Tunisia and the 40,000 screaming Tunisian fans expected to pack out the stadium.
Arnold must now select between Thomas Deng, Milos Degenek and Fran Karacic to fill the right-back vacancy.
In positive news for the Socceroos, Ajdin Hrustic is expected to join the team to take on Tunisia.
How to watch Socceroos v Tunisia LIVE and FREE on SBS
Saturday, November 26
Group D - Tunisia v Australia
7:30pm - 11:30pm (AEDT) - kickoff at 09.00pm
LIVE on SBS and SBS On Demand
Catch up on France v Australia the way you want with the full replay and 3, 10 and 25 minute highlights all available on the SBS On Demand Match Hub.