Almost 50,000 animals, including rats, fish, horses and dogs were used, in research projects at the University of Newcastle over a three year period.
The vast majority of the animals were rats and mice, which were destroyed upon the completion of the research.
While animal welfare organisations condemn the 'industrial scale' use of animals by universities, the University of Newcastle argued animal-based research had assisted in key breakthroughs in several areas of medical research.
"We understand that involving animals in research can be a challenging subject, however it remains key to answering biological questions that help researchers find new and better ways to treat disease, addiction and illness," Deborah Hodgson, Acting Deputy Vice-Chancellor (Research and Innovation) said.
"Animal models are helping our researchers to develop better treatments for ovarian cancer, pancreatic cancer, leukemia, dementia, Alzheimer's disease, drug addiction, lung disease, HIV, malaria and countless other serious medical conditions."
University research data shows 13,212 animals were used in 2016, 18,260 were used in 2017 and 16,371 were used in 2018.
Data for 2019 will not be available until May this year.
Fish, amphibians, reptiles, Indian Mynah birds, rabbits and guinea pigs were listed among the animals used in experiments.
In addition, researchers also work with animals in field studies, such as equine breeding or improving care for horses and dogs with skin cancer.
All research involving animals is approved by an independent animal care and ethics committee.
Researchers are also required to conduct their research in accordance with the Australian code for the care and use of animals for scientific purposes.
The university said many of the research projects involved the collection of samples from animals such as urine, faeces and blood.
Once collected the animals continue to live in their natural environment.
An RSPCA spokesman said the organisation was opposed to the use of animals in experiments or procedures that cause "pain, suffering, distress or lasting harm".'
"This includes aspects such as fear, hunger, boredom, social isolation, and other forms of distress. Research animals must be treated with respect and not merely as objects or scientific tools," he said.
"RSPCA Australia is also opposed to the use of animals in the testing of non-essential substances such as cosmetics, tobacco, alcohol and household products."
New legislation prohibiting the use of animals in the testing of some types of cosmetic ingredients will come into effect from July 2020.
It follows intense worldwide pressure to reduce or stop the use of animals in scientific testing.
Professor Hodgson said emerging technologies and the university's commitment to 'reduce, refine and replace' principles meant the university was reducing the need to use animals.
"Collaborative work between laboratory-based researchers and mathematical modellers (or biostatisticians) is helping improve the overall design of studies, so that only the minimum numbers of animals required are used to produce relevant, high-quality and representative data," she said.
"Researchers are also benefiting from advanced computer modelling and mathematical algorithms that detect patterns and predict outcomes, which can lead to fewer animals required for some studies."
"We're committed to having strong community involvement on our Animal Care and Ethics Committee - an independent committee which oversees the care and welfare of animals we work with. It is made up of community members, animal welfare representatives, veterinary scientists and academics. Having a breadth of perspectives is important."
But co-founder of Newcastle-based Save Animals in Laboratories Pauline Groenland said remained sceptical.
"It is very commonly known that the failure rate of animal research is higher than 90 per cent with many scientists noting that the small amount of success is "in spite of research"," Ms Groenland," who participates protests against animal testing outside the university, said.
"So over a century of animal research and the success rate has not risen yet non-animal research has already a proven success rate of 85 per cent."