Rabbits have been found inside the predator-proof Mulligans Flat Woodland Sanctuary, prompting authorities to urge visitors to always check that gates are closed behind them.
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The sanctuary's electrified fences provide safety from foxes and feral cats for its population of native Australian species, including echidnas, wallabies, shingleback lizards and quolls.
It is also home to about 600 eastern long-neck turtles who are set to help deal with Canberra's carp problem.
But now, sanctuary manager Will Batson has said an unchecked rabbit population inside the reserve would compete with the native species for food and disturb the groundcover.
"They just degrade the whole system," Mr Batson said.
Mr Batson said rabbits, without predators to keep their numbers in check, could breed at an exponential rate.
One female rabbit was able to give birth to 50 kits a year.
The most likely way they entered was through an open gate.
"It's really important people double check," he said.
The sanctuary is now closed from 6pm to 6am, Monday to Thursday until October as rangers use guns to hunt rabbits.
"It's the least risky method, because of the number of native species we've got in there we're really restricted in what techniques we can use," Mr Batson said.
"Baiting and poisoning is the most effective method for controlling rabbits but because of the impacts on the rest of the native species we don't use that."
Rangers were reluctant to rip up rabbit warrens because native quolls would use them as nests.
A quick response was necessary.
"The best chances are when numbers are low," Mr Batson said.
Despite the possibility that people were responsible for letting the rabbits in, Mr Batson said there were no plans to close the sanctuary to the public permanently.
Members of the public are asked to report rabbit sightings to admin@woodlandsandwetlands.org.au