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One of the main characters is nonbinary, and states they aren't male or female.
One of the side characters is canonically agender.
A character who's part of the main cast, Osana Najimi, is canonically genderfluid.
An important member of the main cast, Kei, canonically states that they "gave up being a man" "but it's not like that means they want to be a woman either." So, nonbinary!
Generally lots of good discussions about gender and intersex topics for aging adults, but also the protag is, from what I can understand, a transmasc intersex enby.
The love interest of this smutty romcom is probably somewhere in the intersection of bigender & genderfluid. Their body changes physical sex periodically, which it wouldn't do it if they really wanted to stick to one gender or another.
This one's complicated - the audience is never given confirmation on what the main love interest's gender is from the character themselves, binary or not. But they also haven't told anyone in-universe either, except their partner bc it's gonna come up anyways, so they're happy being treated by everyone as enby & I think that's good enough.
The primary love interest is canonically a nonbinary intersex person (although their intersex status is caused by magic). They've canonically stated they're neither a boy or a girl.
It's made slightly more apparent in the anime adaptation but it's still pretty clear in the manga that Kino, the protag, doesn't consider themselves male or female. When addressed by gendered terms they sometimes correct the other by saying "please don't call me that, I'm just Kino"
The protagonist has stated they aren't a boy or a girl but also seems to feel like one binary gender or another sometimes, so I'd tentatively put them down as being genderfluid.
Fukakai na Boku no Subete o