Already having overcome a side strain in order to face India at the Gabba, Hazlewood felt soreness in his calf on Tuesday morning.
Team medical staff say Hazlewood first experienced "awareness" in the calf while warming up for day four, prompting him to stay off the field for the start of play.
He remained absent for two overs, before coming on and bowling just before the first drinks break.
The seamer was notably down on pace in that over, with his first delivery an uncharacteristic long-hop that KL Rahul dispatched to the point boundary.
When drinks were taken Hazlewood was in deep discussion with captain Pat Cummins, Steve Smith and Australia's team physio, before leaving the field.
Cricket Australia have since confirmed he will be taken for scans to assess the issue.
An Australian team spokesperson said Hazlewood was suffering from "calf awareness" with the fast bowler set to undergo medical scans to determine the extent of the injury.— cricket.com.au (@cricketcomau) #AUSvIND https://t.co/ooW7SaikHmDecember 17, 2024
Any injury to Hazlewood could have serious ramifications for the rest of the series.
Australia are eying off the chance to enforce the follow on against India in Brisbane, with rain having taken a significant portion out of the game.
That is likely to leave Mitch Marsh needing to bowl more overs than expected, after having battled his own injury issues this summer.
Scott Boland would loom as Hazlewood's replacement if he is to miss any further Tests, after already filling in for him in Adelaide.
But any strain on the workload of Cummins, Starc or Marsh could prompt Australia to have to blood a debutant, with Sean Abbott, Brendan Doggett and Beau Webster all part of extended squads this summer.
A week-long gap is scheduled between the Brisbane and Melbourne Tests, with only three days separating the end at the MCG and start in Sydney.
Hazlewood has battled side strain issues in the past, with similar injuries largely responsible for him playing only three Tests between the end of January 2021 and start of June 2023.
The quick had bowled five overs on Monday for Australia without issue, before Tuesday's concern.
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