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.

Doña Bárbara

Doña Bárbara (BAHR-bahr-ah), a beautiful but unscrupulous mestiza, once wronged by a white man and now taking her revenge on all men. She is superstitious and given to witchcraft. For a time she was the mistress of Lorenzo Barquero, heir to half the Altamira ranch, and by him had a daughter, Marisela. Then she ran him off his land and took possession. With the help of her cowboys, she is acquiring the rest of the ranch by moving the boundary fences and stealing the cattle. She has won over the local authorities. Unfortunately, she falls in love with Santos Luzardo, heir to the other half of the estate. When she goes soft, her followers desert her. Finally she rides to Altamira to shoot her daughter, whom she considers her rival, only to soften at the sight of Santos’ display of affection for Marisela. She draws up papers leaving the Barquero land to Marisela and restoring what she has been stealing, then rides off, never to be heard of again.

Dr. Santos Luzardo

Dr. Santos Luzardo (SAHN-tohs lew-SAHR-doh), a descendant of owners of the other half of the ranch. Taken to Caracas as a child, he studies law, then returns to his ancestral property in the wildest section of the Arauca River basin of Venezuela in order to get the land ready for sale. Seeing its deterioration under irresponsible overseers, he determines to restore the ranch to productivity. To end the long feud between the Luzardos and the Barqueros, he brings the dying Lorenzo Barquero and his daughter Marisela from their swamp cabin to live at the Altamira ranch house. From his knowledge of law, he is able to force the magistrate to call for a round-up to separate the cattle. He also sends some of his cowboys to collect heron feathers, from whose sale he will get money to repair his fences. They are murdered and the feathers stolen. When the local magistrate does nothing, Santos decides to follow the law of the jungle and match violence with violence. Hunting the feathers, on a tip from Bárbara, he finds the Wizard, her most trusted henchman, and leaves him dead.

Lorenzo Barquero

Lorenzo Barquero (loh-REHN-soh bahr-KEH-roh), the weak-willed heir to half the Altamira ranch. Doña Bárbara becomes his mistress and gives him a daughter, Marisela. Tiring of him, Bárbara drives him off his land and begins to take over the ranch.

Marisela

Marisela (mahr-ee-SEH-lah), a young girl, beautiful under her dirt, the daughter of Lorenzo Barquero. Her mother Bárbara abandons her, and her father sells her to William Danger for five bottles of whiskey. But Santos determines to civilize her. She falls so deeply in love with him that she is willing to challenge her mother’s witchcraft in order to get him for herself. Finally they marry, and the two halves of the estate are reunited.

Señora Luzardo

Señora Luzardo, the mother of Santos, whose life has been tragic. Not only has she seen the constant feuding between the two branches of the family, but her husband killed her oldest son in a quarrel and in remorse starved himself to death. She takes Santos back to the civilization of Caracas to study, but she cannot keep him there.

William Danger

William Danger, a North American squatter who enjoys demoralizing Lorenzo Barquero by giving him liquor. He is compelled by Santos to build fences, since his herd is too small to run wild.

Antonio Sandoval

Antonio Sandoval (SAHN-doh-vahl), a cowboy faithful to Santos who helps him rebuild the ranch.

Balbino Paiba

Balbino Paiba (bahl-BEE-noh PI-bah), the Luzardo overseer, whose love for Doña Bárbara makes him unfaithful to his employers. To make money, he steals feathers and kills Santos’ cowboys. He is killed at Bárbara’s orders when she tires of him and prefers Santos Luzardo.

Carmelito López

Carmelito López (kahr-mehl-EE-toh LOH-pehs), one of Santos’ cowboys, killed by Balbino.

Pajarote

Pajarote (pah-hah-ROH-teh), a cowboy who helps Santos.

Ambrosio

Ambrosio (ahm-BROH-see-oh), a one-eyed cowboy musician.

Melquíades Gamarra

Melquíades Gamarra (mehl-KEE-ah-dehs gah-mah-rrah), called The Wizard, Bárbara’s trigger man.

Juan Primito

Juan Primito (hwahn pree-MEE-toh), a demented cowboy who feeds the birds of ill omen and prophesies evil.

Ño Pernalete

Ño Pernalete (nyoh PEHR-nah-leh-teh), the local magistrate, partial at first to Doña Bárbara, but compelled by Santos to administer justice.

Mujiquita

Mujiquita (mew-hee-KEE-tah), the clerk of the court.