Doña Bárbara is the vengeful mestiza who is suspected to be in involved with witchcraft. She is ruthless and power hungry. She had a child, Marisela, in her affair with Lorenzo Barquero. In her attempt to continue expanding the boundaries of the stolen land from Barquero, she falls in love with the other heir of the Altamira ranch, Santos Luzardo. Her ruthless control is softened when Luzardo negotiates boundaries and terms for their respective lands. Doña Bárbara is estranged and detached from her daughter. However, her love for Luzardo wins as she ultimately disappears and leaves the deed in her daughter's name.

Santos Luzardo is the heir to part of the Altamira ranch. He is raised in Caracas as a child and separated from his familial property by his mother. His attendance at law school helps him maneuver the legal aspect of the repossession of his property in the midst of the chaos with Doña Bárbara. He is careful and thoughtful in his actions to regain the property. First, he squashes the long standing feud between his family name and the Barqueros. He is then able to round-up and separate the cattle that have been stolen by Doña Bárbara. He resorts to murder once his mission to finance his fence building is thwarted by one of Doña Bárbara's henchmen. He ultimately marries Marisela and rejoins the Altamira ranch under one name.

Lorenzo Barquero is the other owner to part of the Altamira ranch. He has a child with his mistress who soon leaves him. He is driven off his property and lives in a swamp cabin until Luzardo resolves their family feud. He lives with Luzardo and Marisela until his death.

Marisela is the daughter of Lorenzo Barquero and Doña Bárbara. Her youth and beauty are mistreated by her parents. Her mother abandons her and her father sells her to William Danger. She moves to Luzardo's estate once he meets with her and her father to resolve the family feud. She falls in love with Luzardo and challenges her mother for his affections. She ultimately marries Luzardo and reunites the ranch under one name.

Señora Luzardo is the mother of Santos. After the loss of her eldest son in the family feuds, she takes Santos to Caracas to escape from the violence.

William Danger is a squatter who takes advantage of Lorenzo Barquero by giving him liquor. He is able to negotiate buying Marisela by capitalizing off of Lorenzo's bad habits.

Antonio Sandoval is a faithful cowboy to Santos as he helps him rebuild the ranch.

Balbina Paibo is the overseer and lover to Doña Bárbara. He thwarts Santos's plans to sell feathers to finance his fence. He is killed once Doña Bárbara falls in love with Santos.

Melquíades Gamarra, also known as "The Wizard," is a henchmen of Bárbara’s. He is falsely identified by Bárbara as the thief who stole Santos's feathers. He is killed by Santos in revenge.

Carmelito López, Pajarote, Ambrosio, and Juan Primito are cowboys.

Ño Pernalete is the local magistrate who was won over by Bárbara at first. He is reminded of his position to follow legal procedure by Santos.