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Edit: layouts are "staged" a little bit around key and item collection in biome 1 specifically so if you beat biome 1 the first time you unlocked the boss door and then didn't spend much time repeating it, you never saw the regular difficulty version of that biome.
Aside from Malformed enemies that were mentioned, but they appear to be another thing that shows up in earlier areas as they're encountered because I haven't seen any difference in their spawns before/after beating the game.
I distinctly remember being killed by a malformed severed with the purple laser walls in a Biome 1 secret room, during my run back to Nemesis.