Castlevania features multiple LGBTQ+ characters, including the openly lesbian vampire generals Striga and Morana, whose loving relationship is portrayed with dignity and complexity. Alucard's bisexuality is explored through a tragic polyamorous relationship, but the narrative frames the betrayal as external, not a condemnation of his identity. While some queer characters are antagonists, their identities are not the source of their villainy, contributing to a net positive and affirming portrayal.

Isaac, a character from the Castlevania video game series, was depicted with dark skin in the source material but without a specified race. The animated series portrays him as explicitly Black, which constitutes a race swap.

The film consistently portrays the organized Christian Church as a corrupt, ignorant, and oppressive institution. Its fanatical actions, particularly the burning of Lisa, directly instigate the main conflict, and its clergy are depicted as antagonists who hinder efforts to save humanity.