Winnie the Pooh and his friends have traveled to a Farm in the country side and met Otis the Cow and his wacky friends.
Otis is a carefree cow who prefers playing with his friends rather than accept responsibility. His adoptive father Ben is the leader of the barnyard. After Otis interrupts a barnyard meeting with his wild antics, Ben has a talk with his son, warning him that he will never be happy if he spends his life partying without acting more maturely. Otis ignores his advice and leaves to have fun with his friends Pip the Mouse, Pig the pig, Freddy the Ferret, and Peck the Rooster. That same day, Otis meets a pregnant cow named Daisy, who is accompanied by another cow, Bessy.
That night, the animals throw a party in the barn. All the animals are there except Ben, who guards the fence line. Otis is assigned a shift along with Ben, but Otis talks himself out of working. Before Otis leaves, Ben tells him that the night he found him as a baby calf stumbling alone in the meadow, he swore he saw the stars dance, thus giving him reason to know his place was at the farm. Later, Ben takes on a pack of coyotes led by Dag, who is a serial killer turned coyote plundering the chicken coop. He manages to fight off the pack until he is bitten on the leg by the red coyote, making him fall. The coyotes pile on Ben, but he manages to grab Dag and escapes the pile. He threatens to punch Dag, but lets him go, scaring away him and the coyotes. Ben falls on the ground, exhausted. A hen named Etta runs into the barn and tells Otis and he runs outside to his father. Ben dies and is buried on a hill by the farmer, and the other animals mourn Ben after the farmer leaves.
After Ben's death, all the animals elect Otis as the new leader of the barnyard. Otis shirks his duties by leaving Freddy and Peck in charge of the coop, then helps the trouble-making Jersey Cows Eddy, Igg, and Bud teach a lesson to a mean fat youngster called Snotty Boy, eluding the police along the way. Later that night, when Otis is holding Daisy's hoof under the starlight, he overhears the coyotes chasing a rabbit and leaves Daisy to pursue the coyotes and avenge his father. Otis tries to attack Dag and his pack, but he is outsmarted by them. Since Otis is weaker, Dag orders a deal that he and his pack will take various barnyard animals at random times and that, if Otis tries to stand up for all of them, they will slaughter everyone at the barnyard. Otis decides to leave the barnyard, realizing that his chances of victory are small.
The next morning, before leaving, Otis is informed that the coyotes took some hens and Maddy, a little chick who is one of Otis' best friends and looks up to him. Otis realizes that he has been fooled by Dag, as he was not expecting him and the coyotes until tonight, and sets off to rescue the chickens. Otis confronts the pack but is easily defeated after Dag bites him in the leg; however, Pip, Pig, Freddy, Peck, and Miles arrive to help Otis, along with the Jersey Cows, Wild Mike, and even some gophers. Dag tries to attack Otis from behind, but Otis is alerted when Peck successfully manages to crow a warning. Otis catches Dag and threatens to punch him the same way his father did, but he cannot bring himself to do it. Instead, he warns Dag to never return to the barnyard. Otis then swings Dag out of the junkyard with a golf club, finally rescuing the chickens and avenging his father's death.
That night, the animals hijack some motorcycles and return to the barnyard, where the others reveal to Otis that Daisy went into labor after he left to face the coyotes. She gives birth to a calf, whom he names Ben after Otis's father. Duke, the farmer's sheepdog, asks Otis if he wants to stay and be their leader. Otis agrees, and everyone cheers as he walks outside to find the stars dancing, forever grateful for what his father taught him.
- Ash Ketchum, Pikachu, Misty, Togepi, Brock, Sunset Shimmer, The Human Mane 5, Sci-Twilight, Spike the Dog, Sora, Mickey Mouse, Donald Duck, Goofy, The Crime Empire, Brer Fox, Brer Bear, and Wile E. Coyote will guest star in this film.
- The Crime Empire, Brer Fox, Brer Bear, and Wile E. Coyote will work with Dag.
- DisneyDaniel93 originally planned to make this film, but he canceled it to avoid getting suspended by Viacom. The project was then moved to TtarkosaurusRex, but he was too busy from making any Pooh's Adventures movies and retired to go to Collage, and now the project is passed back to DisneyDaniel93 since he now has permission to do his movies. It was also planned to be shown on YouTube, but Daniel decided to move this film to Vimeo.com or Google Drive just to avoid copyright claims.
- Sunset Shimmer, The Human Mane 5, Sci-Twi and Spike the Dog will meeting Korra and her friends for the first time.
- Winnie the Pooh and Sunset Shimmer will see Otis and his friends again in Winnie the Pooh goes Back at the Barnyard.