Two Mules for Sister Sara is a Western film that does not feature any identifiable LGBTQ+ characters or themes. The narrative centers on a male and female lead, with no elements suggesting queer identity or relationships are present in the story.

This film does not include any identifiable transsexual characters or themes within its plot or character arcs. The story centers on a male mercenary and a female character posing as a nun. Therefore, the framework's evaluation of transsexual portrayal is not applicable.

The film features Sister Sara, who is involved in the conflict against French forces. While she contributes significantly through her knowledge of explosives and strategic guidance, she does not engage in or win any close-quarters physical combat against male opponents. Her actions are primarily supportive or involve ranged tactics.

Two Mules for Sister Sara is an original story, not an adaptation of pre-existing material or a biopic. Its characters were created for this film, thus there is no prior canonical or historical gender to swap.

The film "Two Mules for Sister Sara" is an original story, not an adaptation of pre-existing material with established character races or based on historical figures. Therefore, no character's race was changed from a prior canonical or historical depiction.