Tristar has refused to comment on the future of its Wodonga clinic, amid the departure of a physician and the closure of another site.
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Tristar Medical Group's director of clinical operations Anne Gardner did not respond to repeated requests for comment on Wodonga's future in light of Dr Chaudhry Mansoor Ahmed's departure.
Receptionists at the company's head office said Ms Gardner was not taking calls.
At least two other physicians have left Tristar Wodonga in the past 12 months.
It comes as Tristar Medical Group in Portland prepares to close.
Portland District Health's Chris Giles told The Standard the Tristar clinic previously had three full-time general practitioners.
"That's gradually gone from three to two, two to one and now it's closing," she said.
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For months, the Wodonga clinic has been operating with just two doctors, including Dr Chaudhry Mansoor Ahmed, with signs by the entrance informing patients the clinic was not running at full capacity.
In mid to late August, the clinic cut its operating hours from 8am to 8pm weekdays and 10am to 6pm weekends, to 8am to 3pm Monday to Friday and 10am to 2pm every second Saturday and Sunday.
A sign on the door informing patients of the change said "due to doctor shortages our clinic will now be closing early until further notice".
Online bookings for the Wodonga Tristar clinic no longer work, with the website simply saying there were 'no appointments available' for any selection.
Appointments can still be made by phone.
In July, Ms Gardner told The Border Mail Tristar Medical Group was facing 'significant challenges' but the Wodonga and West Albury clinics had survived a business restructure.
At the time Tristar said two GPs were due to start in Wodonga "in coming months".
Since then multiple doctors have accused Tristar of underpaying them.