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Amalthea lives in a magical forest in which each animal is protected from hunters. While she has not met any other unicorns, she believes there must be others like her out there.

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Amalthea learns from two hunters and a butterfly that she is the last of her kind since an evil Red Bull has herded unicorns to the ends of the earth. Amalthea refuses to believe that and begins her journeys to find them.

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After being captured by the witch, Mommy Fortuna, Amalthea meets and befriends Schmendrick, an incompetent wizard in Mommy Fortune's service. With his help, Amalthea is freed and Fortuna is killed by a lion once freed by Amalthea. Schmendrick and Amalthea continue on together and meet Molly Grue. Together, Molly and Schmendrick help guide Amalthea on her journey.

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Before Amalthea can continue on, she encounters the red bull. Before she can be captured by it, Schmendrick uses his unpredictable magic and transforms her into a woman. The bull loses interest in her and leaves. Amalthea is shocked by the sensation of mortality and brought to tears. Schmendrick promises to return her to her original form after her quest is complete.

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By this point in the movie Amalthea has met the talking butterfly, who started her on her quest, a farmer, who could not see her unicorn horn and tried to capture her to sell her, Mommy Fortuna, who captured her for display, Schmendrick, who freed her, and Molly, who had an extreme emotional attachment to her on sight. Amalthea has also met her main enemy, the red bull which tries to capture her and fails. And lastly, she has passed through three trials: the farmer, the witch, and a first encounter with the red bull.

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They follow the bull and find Haggard's castle. This is where Amalthea actually receives her name when Schmendrick introduces her to Haggard. Haggard agrees to lodge the trio and soon after Amalthea meets Prince Lír. Molly eventually learns the location of the Red Bull's lair from the castle's cat.

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Amalthea struggles internally due to her newly found human emotions. She begins to forget her true self and falls in love with Prince Lír, and considers abandoning her quest in favor of mortal love. Haggard confronts Amalthea, hinting at the location of the unicorns. Yet the magic hidden in her eyes has him doubting that she's as simple as she appears.

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Molly, Schmendrick, and Amalthea are joined by Lír as they enter the bull's den, and are trapped there by Haggard. Schmendrick explains to Lír what they are looking for and reveals Amalthea's true identity. Lír admits that he loves her. The Red Bull appears, no longer deceived by Amalthea's human form, and chases after her. Schmendrick turns Amalthea back into a Unicorn, but she is unwilling to leave Lír's side. The Bull begins driving her toward the ocean just as it had done to the others before her. Lír tries defending her, but is killed by the Bull. Enraged, Amalthea turns on the Bull and forces it into the sea. As the beast is engulfed by the water, the missing hundreds of unicorns emerge from the raging sea.

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With the release of the thousands of missing unicorns, Haggard's castle collapses into the sea, and Haggard, watching all of it, falls to his death. On the beach, Amalthea magically revives Lír before she leaves him. Schmendrick assures Lír he gained much by winning the love of a unicorn, even if he is now alone. Amalthea later says goodbye to Schmendrick, who admits he wronged her by burdening her with the taint of mortality. She disagrees about the importance of his actions, as they had helped restore unicorns to the world and made her experience regret and love. Schmendrick and Molly watch Amalthea depart for her forest home.

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Amalthea begins her long journey home with the rest of her kind all the way back to the magical forest. Her prize is the gift of feeling human emotions and the pleasure and comfort of knowing she isn't the last unicorn.