Staff at Blair Drummond Safari Park near Stirling called the emergency services just before 9pm yesterday after the fire broke out in the bird of prey centre.
Fire crews raced to the blaze, which is thought to have been sparked by a faulty heating system, from Stirling and nearby Doune.
The fire was brought under control by 10pm.
But safari park officials said the two much-loved birds of prey, including the park’s giant male Rüppell’s Griffon vulture, were sadly dead.
Other raptors were successfully moved to other areas of the park as the raptor centre was closed to visitors today.
A spokeswoman said: “We are sad to announce that two birds of prey have died in a fire at Blair Drummond Safari Park.
“The fire, which we believe was caused by a fault with our heating system, started in the corner section of the aviary in our bird of prey centre.
“On-site safari park staff reacted immediately. The rapid response from Scottish Fire and Rescue ensured that the fire was quickly contained and extinguished.
“The fire has destroyed two aviaries and a store room in our bird of prey centre. Unfortunately, there was nothing that could be done to save the birds in these aviaries and a Ruppell Griffon vulture and a white tailed Sea eagle both died.
“Our vet has checked the other animals in this section and we will continue to monitor their health. Early signs suggest that they have not been affected.”
She added: “Our bird of prey keepers Callan, Dom and Mark have a very close bond with the animals they look after and are understandable very upset at this event, as we all are.”
The dead birds were among the largest anywhere in Scotland, with wingspans of more than eight feet.
Last year, Blair Drummond hoped to become the first collection in Scotland ever to breed the endangered Rüppell’s Griffon vulture, which has suffered a 95 per cent decline in its native central Africa in the past 20 years.
It was hoped the resident vulture’s descendants might be returned to the wild to help the species recover or repopulate.
The Scottish Fire and Rescue Service (SFRS) said Operations Control quickly mobilised three fire engines to the aviaries inside the park where firefighters extinguished the fire.
SFRS station manager Alex McCutcheon said: “The crews responded quickly and effectively to this incident.
“On arrival the fire had already taken hold of two aviaries, but quick and decisive action ensured that the fire was contained to stop further fire spread and extinguished.
“Safari park staff and crews managed to save the majority of the birds and relocate them but sadly two did not survive.
“I would like to thank the firefighters involved for bringing this incident to a swift conclusion and also the staff at the park for their assistance.”
The bird of prey centre is expected to reopen tomorrow.
The safari park spokeswoman added: “This will have no effect on the rest of the Park and our events over the Easter weekend will go ahead as planned.
“We would like to thank the Scottish Fire and Rescue service for their swift response. We would also like to thank our visitors for their concern and support.”
The fire broke out just days after the safari park reopened to the public on 17 March. The previous day, staff were put through fire drills in a “realistic training exercise” centred around the lion enclosure.
Community firefighter Heather Steel from the Scottish Fire and Rescue Service said at the time: “A fire at the safari park is a rare occurrence, but it is vitally important that Blair Drummond Safari Park know exactly what to do if such an emergency arose.”
Police Scotland said the fire last night was “non-suspicious”.