Voiced by: Chris HardwickOtis is the leader of the barnyard. He is a cow.
Voiced by: Jeff GarciaOtis' bestfriend, a mouse.
Voiced by: Leigh Allyn BakerA female cow at the barn who has a crush on Otis and vice versa, though they never get to admit their feelings for each other.
Voiced by: Tino InsanaA pig who lives at the farm and hangs out with the main characters.
Voiced by: Cam ClarkeA ferret who lives at the barn. Best friends with Peck the rooster, and constantly scold himself for thinking about eating him or any other chicken on the barn.
Voiced by: Rob PaulsenA rooster who lives at the barn. He's best friends with Freddy the Ferret.
Voiced by: Wanda SykesA cow who lives on the farm and criticizes Otis.
Voiced by: Maria BamfordA human who lives close to the farmer. She knows that the animals walk and talk, and is always looking for evidence, causing her husband to believe she's crazy.
Voiced by: Steve OedekerkMrs. Beady's nephew who likes to mess with the barn animals.
Voiced by: Rob Paulsen and Steve OedekerkTwo guys who work at a pizza restaurant and pop up to say stupid things.
Voiced by: Fred TatascioreThe owner of the farm. He's not a very important character, and he's unaware of the animal's ability to walk and talk.