The difference between Third gender and Two-spirit
When used as nouns, third gender means a category (a gender), present in societies which recognize three or more genders, which is neither male nor female, whereas two-spirit means a native (north) american gender-variant, homosexual or bisexual person (especially one belonging to a traditional tribal third-gender, fourth-gender or transgender cultural category that has a ceremonial role).
Two-spirit is also adjective with the meaning: pertaining to or being a two-spirit.
check bellow for the other definitions of Third gender and Two-spirit
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Third gender as a noun (anthropology):
A category (a gender), present in societies which recognize three or more genders, which is neither male nor female; in some societies, the category may contain individuals who are intersex or androgynous, while other societies may categorize transgender people into it.
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Two-spirit as a noun:
A Native (North) American gender-variant, homosexual or bisexual person (especially one belonging to a traditional tribal third-gender, fourth-gender or transgender cultural category that has a ceremonial role).
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Two-spirit as an adjective:
Pertaining to or being a two-spirit.
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Two-spirit as an adjective (theology):
Involving two spirits; especially, pertaining to the doctrine of dualism espoused in the so-called Treatise on the Two Spirits in the Dead Sea Scrolls.