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Answer for the clue "Second letters of this puzzle's five theme answers (seen in order) ", 5 letters:
aeiou

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Wikipedia Word definitions in Wikipedia
AEIOU may refer to: a, e, i, o, u, a traditional list of vowel letters in the Roman alphabet A.E.I.O.U. , a device used by the Habsburgs aeiou Encyclopedia , a free online collection of reference works in German and English about Austria-related topics ...