The property, 755 Concord St., straddles the Holliston-Ashland line.
The Metrowest Daily News reports some 60 animals were inside the barn when the fire broke out.
A Sunday morning fire inside a barn in Holliston killed several animals living inside the structure. The fire was knocked down within two hours, but the barn was "totally destroyed", he said in a phone interview.
The Animal Rescue League of Boston is helping to care for the surviving animals.
A number of officials were called to investigate after the fire, including an Ashland/Holliston Animal Control officer, a Holliston building inspector, a Board of Health official, a wiring inspector, and a State Fire Marshal investigator.
Officials said they are looking into what started the fire.