A tourism operator says a late start to the breeding season of little penguins on Granite Island could indicate the end of the population.
Mating season for the birds usually begins in July, but Dorothy Longden from the island's penguin centre says they are yet to find any eggs at key nesting sites.
A census last year counted only 26 penguins on the island, down from 1800 twelve years ago, and prompted a warning from ecologists the colony was on the brink of disappearing.
The birds are a major tourist drawcard on the island off Victor Harbor, south of Adelaide.
Ms Longden says the number of penguins outside of captivity remains drastically low.
"Each year we've been hoping that all the numbers would pick up but once the colony starts to go, then it goes down rather fast and this is what's happened this year," she said.
"From 26 penguins that were counted last year to... three sightings on the penguin tours in the evening.
"There is just no evidence of any eggs at all.
"If there's no chicks this year, then we won't have a colony next year. I mean, we haven't got a colony now."
Ms Longden says fortnightly inspections of nesting sites have revealed only abandoned nests and hopes the ten penguins in captivity can breed to restore the colony.
A census of the penguin population is expected to be carried out next month.
The cause of decline remains unknown but has previously been blamed on predators including cats, dogs, foxes and seals.
Ms Longden says two eggs have been laid inside the centre's enclosure, but neither have hatched and there could be a problem with the birds' fertility.
"Charlie the male, he's had two different partners over the last few years and with each different partner, they've successfully had eggs but nothing's happened to them," she said.
The penguin center cut back its tours earlier this year because of the declining population.
Last year, an ecologist said dozens of the creatures had washed up dead on beaches in the past 12 months.
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