Tula protected Middle Island's penguin colony for over eight years, along with her sister Eudy who passed just five months ago at the age of 12.

The pair were the first dogs specially trained to protect Warrnambool's Little Penguin Colony. Tula retired in 2019 and had since played an integral part in training younger dogs.

Lead dog handler Dr Trish Corbett said Tula would be deeply missed.

"Tula was the lead guardian - she was always the first to jump up and bark if there was a threat," she said.

"Middle Island was her domain. She had such a strong instinct and loved protecting her island.

"Tula, together with Eudy, also did an amazing job in training the new generation of guardians. Even after she retired to the 'farm', she continued to protect the chickens and train the younger dogs. You couldn't tell her not to work. She lived for it."

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Dr Corbett said Tula had arthritis and two cruciate ligament surgeries in recent years.

"She was old and it was her time," she said.

"Tula spent her last night with loved ones visiting her and went to sleep with love and a stomach full of her favourite food.

"I'd like to thank the Vet Group for their compassion and care, especially during Tula's final days. I'd also like to thank Pet Stock for their support in feeding our team of guardians over the years."

Warrnambool mayor Richard Ziegeler thanked Tula for all she'd done.

"With the sad passing of Tula, it's the end of an era for one of the world's most innovative conservation projects," he said.

"It's a poignant after note to the recent death of Eudy, Tula's sister. The Maremmas are iconic in Warrnambool. Part of the main entrance sign for the city depicts penguins in front of Middle Island under the watchful eye of a Maremma dog and the Middle Island Penguin Project is something that fills the people of Warrnambool with a lot of pride.

"The legacy that Tula and her sister Eudy have created will live on in the work undertaken by the next generation of Maremma guardian dogs and the dedicated people who make the project possible.

"I offer my sincere condolences to the team who have worked with these two magnificent dogs over the past 13 years."

Tula's ashes will be scattered along with her sister Eudy's on Middle Island.

A public memorial for the sisters will be live streamed on the Middle Island Maremma Penguin Project Facebook page at 7.30pm on Friday November 5.

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