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In the anime, latter half of the first season, we see that Kumoko saves a baby human vampire from some bandits and later guards her by camping out near the town. We also know that this baby vampire is a reincarnation. (I believe it's Negishi Shouko)

There's an adult Sophia vampire that serves the Demon Lord Ariel. We also find out she's a reincarnation. I thought these were both one and the same, just huge time gaps, but in episode 20 we see adult Sophia heading to attack the town where baby Sophia is, and from what I can tell it's in the same timeline. Are the two Sophia's genuinely different people, connected in some way, or are they the same person and I'm misunderstanding the timelines?

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Yes both are the same person.

Your confusion comes because in episode 20 there is a shift in time that is not super obvious. There was a short transition scene after Kumoko talked to herself about what she was going to do regarding the army heading to Keran County of the Sariella Country. After she made up her mind and left the Tree, it showed the same tree several times but as time passed, going from a thriving tree full of leaves to a seemingly dead tree with no leaves indicating that time had passed. This transition is easily missed. The Army with Sophia, Hugo, and the Demon lord is heading towards the Elf village.

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