The following is a list of characters who appearing in the 2006 film Barnyard and its Nickelodeon animated sequel television series Back at the Barnyard. The main characters in the film are unique in that they are animated as female milking cows, but are voiced over by male actors such as Kevin James and Sam Elliott. A total of 178 individual characters were built by animators for the film.

Overall main characters

Otis (voiced by Kevin James in the film and Chris Hardwick in the series) is a male Holstein Friesian and the leader of the Barnyard. He is carefree, goofy and energetic, lacking responsibility and goofing off at any given opportunity. Despite this, Otis is highly intelligent and clever enough to come up with solutions to a problem and cares deeply about his friends and family. His superhero alter-ego is "Cowman". Despite being a bull, he has an udder.

Pip Pip (voiced by Jeffrey Garcia) is a slight Mexican-accented wisecracking barn mouse who is Otis' best friend. He has a crush on Bessy, the sassy and sarcastic cow best friend of Abby, but she always rejects him. PiP’s superhero alter-ego is Ratboy, the sidekick of Cowman. Pip is not usually involved with the gang's crazy schemes as much as Freddy, Peck, Pig, etc. are, but still, he usually comes along for the ride. Pip’s human disguise makes people question him, to which another animal says, 'He's the world's smallest man!'.

Freddy Freddy (voiced by Cam Clarke) is a dimwitted ferret who is best friends with Peck, a rooster whom he's often tempted to eat; he used to be a carnivore, but then he's become a vegetarian to assimilate to the barnyard. However, in multiple times in the series, Freddy has shown gestures of or attempted to eat Peck, but every time when he tries, his attempt is thwarted, either intentionally or unintentionally. He is illiterate and has paranoid delusions and multiple personality disorders. It's notable how Freddy is the only character to almost be banished from the barnyard twice for crimes he never committed. Once for seemingly eating Peck when he was really trying to get some cinnamon buns Peck was going to trade for anti-itch cream and the other being framed for attacking his friends when it was really termites possessing a ventriloquist dummy. Also, he's a character who's been struck by lightning multiple times throughout the series, but once, he temporarily became a genius after being struck. Freddy's superhero alter-ego is "Paranoid Man".

Peck Peck (voiced by Rob Paulsen) is an intelligent, but weak rooster who is best friends with Freddy and appears to be unaware of Freddy's recurring urges to eat him. He is considered to be extremely accident-prone, usually seen with a meteorite landing on him. Peck also explains the scientific parts to the audience to explain certain things that have happened. Peck seems to share a slight rivalry with Root, a larger, braver rooster on the barnyard, such as in "Pecky Suave", where we learn that Peck is shown to have a crush on a beautiful hen named Hanna. Peck's superhero alter-ego is "The Green Rooster".

Pig Pig (voiced by Tino Insana) is a filthy and fat but well-meaning and friendly Danish Landrace pig and is the only character in the show to be named after his own species. He is very gluttonous and would eat anything, and he does so uncontrollably. In several episodes, Pig has various segments on the show that often interrupt the episode at its most important parts, with a few memorable segments are "Viewer Mail", "Ask Dr. Pig", a makeover segment and "Hollywood: Fact or Fiction?", which serves as filler time when an episode story is slightly shorter than its airtime. Pig has a strong, but quite strange, love for unicorns, and he wishes for one in the Christmas special, and is also the animal on the barnyard to dress up and disguise as humans the most times. He has a pet skunk named Skunky, and in one episode he is shown to sing in an angelic tenor. In another episode, after Pig sat on Pip, it left a mark, and the animals believe that he is related to the pet pigs of Prince Fripplehoot, telling by a birthmark shaped like a crown on his backside and is therefore royal, but the stuck-up pigs turn him down. His superhero alter-ego is "Mr. Hamtastic". Pig makes a cameo in the pilot episode of Planet Sheen along with Otis.

Duke Duke (voiced by Dom Irrera) is a Border Collie who is in charge of the farmer's flock of intelligent sheep. Duke is clearly no match for the intellect possessed by the sheep he herds. Duke loves being a leader and being in charge of things, though no other animal wants him to be the leader because he is too irresponsible. This is first shown in the film after Ben dies and Duke wants to take his position as leader, but no other animal except his fellow dogs and a reluctant cat, wants him to take the place of Ben and wants Otis instead. In the series, Duke fancies himself as the barnyard's unofficial safety inspector and makes the barnyard watch a boring safety film once every year. Duke only enjoys dog activities, such as playing ball, so he doesn't hang around with Otis and his friends much, and as a result, sometimes feels lonely and left out.

Bessy Bessy (voiced by Wanda Sykes) is a sassy and coarse Guernsey cow that has a tendency to yell and Abby's best friend (Daisy's in the film). She is not afraid to tell it like it is but ultimately means well and can occasionally be nice. She also knows how to pull a prank. Bessy finds Otis childish and idiotic, so she lives by insulting and assaulting him. Despite Otis being intelligent, she believes him to be dumb. Pip is in love with Bessy, but she always rejects him, despite taking advantage of this sometimes. Bessy is also seen implicating with other animals, such as Freddy. She also disguises herself very little and does not participate in Otis' plans or appreciate his intelligence at times. She had a son who floated away with balloons at a county fair. All she has to remember him by is his cowbell, which, strangely, Otis has because Otis is identical to her son due to their coat color. In addition, she is the principal of the farm's school program and the judge of its court.

Mrs. Beady Noreen "Nora" Beady, more commonly known as Mrs, Beady, (voiced by Maria Bamford), is a housewife who lives close by to the farm. She is fully aware of the animals' anthropomorphism and regularly attempts to expose their antics to the world. She is neurotic and has become obsessed with exposing the animals, whose mischief often harms her. But no one else believes her because everyone else thinks she's insane, and whenever she attempts to expose the animals, they always find a way to sabotage her plan. In the pilot episode, it is revealed that Nora is the aunt of Snotty Boy, another enemy of animals.

Farmer Buyer Farmer Buyer (voiced by Fred Tatasciore) is the vegan owner of the farm for whom Otis cares deeply. Still, when the animals need some time alone to work on a private project (a movie they are trying to make, a theme park they are attempting to build, etc.), that does not stop them from trying to drive him away with a prank phone call. In the film, whenever he sees the animals having human-like abilities, he is always kicked unconscious by Miles.

Other main characters Miles Miles (voiced by Danny Glover) is an old mule who is a good friend of Ben's. When Ben dies, Miles is crucial in mentoring Otis who helps him assume a position of leadership and defeat the coyotes. He is absent in the series, although he makes a brief appearance in the opening.

Ben Ben (voiced by Sam Elliott) is Otis' adoptive father who took him in when he found him in the meadow as a calf, and the former leader of the farm. In the film, he is killed by a pack of evil coyotes after successfully winning the fight with them. He is stricter than Otis, who enjoys partying. Daisy also names her baby calf after him at the end of the movie. Ben has only been mentioned on the show once in the episode "Big Top Barnyard", where Otis says he used to take him to the circus in childhood.

Daisy Daisy (voiced by Courteney Cox) is a female cow and Otis' love interest in the film, before she and Bessy arrived at the farm, she used to live on another farm, had a husband, and was also pregnant with her first calf but a flash flood rolled into the farm. She and Bessy found refuge in a meadow, but when the storm was over, everybody had drowned. Later, the Farmer took her and Bessy, and they met the other animals. At the end of the film, she gives birth to her calf and names him after Otis' adoptive father, Ben. Daisy and her son neither appear nor are mentioned in the TV series; she was replaced with another female cow character named Abby in the series.

Dag Dag (voiced by David Koechner) is a red coyote and the main antagonist in the film. He is the sadistic and underhanded leader of a large pack of coyotes who live in a local junkyard and prey on the farm's hens. Dag and his pack capture several of the chickens, but under Otis' leadership, the animals are able to free the chickens and scare away all the coyotes. Otis, after threatening to punch Dag and ordering him to never return, then proceeds to swing Dag out of the junkyard using his golf skills. Dag and his pack are never seen in the series, likely because of the former fearfully heeding Otis' warning.

Coyotes The Coyotes are Dag's henchmen and the central antagonists in the film. At the end, they are scared away by Otis' friends.

Abby Abby (voiced by Leigh-Allyn Baker) is a female cow. Abby is very athletic and excitable, allergic to petunias, has an obsessive-compulsive addiction for organizing, and loves cats. For some reason, she replaces Daisy, Otis' love interest from the movie. Abby also serves as the series' female protagonist in the episode "Abby & Veronica" where it is revealed she has an older cousin named Veronica who tried manipulating Otis and his friends to get what she wants. Her superhero alter-ego is "Cowgirl".

Supporting characters

Snotty Boy Eugene "Snotty Boy" Goldner (voiced by Steve Oedekerk) is a major antagonist in the franchise. He is Reginald and Serena Beady's son, Nora and Nathan Beady's nephew and Bernard Beady's younger cousin. Snotty Boy's uncle and aunt babysit him when his parents drop him off, much to Nathan's dismay. Snotty Boy is incredibly dimwitted, mean-spirited, selfish, spoiled, unpleasant, greedy, dishonest, bratty, and obnoxious. He enjoys torturing the animals, his best friends, and his uncle, which is the basis of the hatred by them. However, he only bullies his uncle when Mrs. Beady isn't around. He is also rude towards his parents and often mocks them as well and they don't even bother to disown him as they love him so much. He was bullied by his older cousin Bernard Beady when he was younger and he acts the way he does because of him.

Mr. Beady Nathan Randall Beady III, more commonly known as Mr. Beady, (voiced by Steve Oedekerk), is Mrs. Beady's husband who miserably tries to convince his wife that animals do not talk. Mrs. Beady often calls him "Mr. Lump" because he always sits on a chair watching television and rejects his wife's suspicions. He is also shown to be miserable on a semi-constant basis, on some occasions wishing for death. He is mostly seen with a beverage in his hand which appears to be either soda or beer.

Sheep The Sheep (voiced by Chris Hardwick, Leigh-Allyn Baker, Cam Clarke, Rob Paulsen and Jeff Bennett) always hang around together in an eternally close herd and are more intelligent than Duke. They often quote famous literature and make snide comments toward Duke. They seem to be very intelligent and philosophical despite their farmly upbringings.

Three Dogs Three Dogs (voiced by Cam Clarke, Rob Paulsen and Fred Tatasciore) are a Golden Retriever, an American Pit Bull Terrier, and a Dobermann. The trio of dogs wear barbershop quartet skimmer hats, bowties, and sleeve garters.

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