Maybe a group of rogue Pokemon that see Ash as an outsider in their world, even though he's now a Pokemon. They could try to drive him out or cause him trouble because they don't trust his human - influenced behavior. They might be afraid that Ash would bring the ways of humans into their pure Pokemon world.
Another possibility is a mad scientist - type character who is interested in Ash's transformation. This antagonist could try to experiment on Ash to figure out how and why he became a Pokemon, causing a lot of distress for Ash as he tries to avoid being captured and experimented on.
It could be a powerful and evil Pokemon trainer who wants to capture Ash in his new Pokemon form. This trainer might be after Ash because he believes that a human - turned - Pokemon has some special power or value.
It could be a trainer who hates Pikachu and wants to capture all of them. This would put Ash, as a Pikachu fan, in direct conflict with this trainer.
It could be a trainer who has a dislike for Charmander. This trainer might see Ash's new - found love for Charmander as a threat and try to undermine Ash's relationship with his Charmander. For example, this antagonist could try to steal Charmander or set up difficult battles where Charmander is at a disadvantage.
It could be Team Rocket. Since they always cause trouble for Ash in the original series, in this fanfic, they might see Ash's transformation into a Pokémon as an opportunity to capture him and use him for their evil plans.
Maybe a powerful and evil pokemon trainer who has a vendetta against Ash. Now that Ash is in a vulnerable pokemon form, this trainer could try to capture and control him. The trainer might have some dark motives related to using Ash's unique situation to gain power in the pokemon world.
A powerful and greedy Team Rocket member could be the main antagonist. They might be after Diancie because of its rare and valuable powers. They could try to capture Diancie and use it for their own evil purposes, like selling its powers or using it to take over the Pokemon world.
A wealthy and power - hungry trainer who wants to stop Ash from completing his Nuzlocke journey. This trainer believes that the Nuzlocke rules are too restrictive and wants to prove that his way of training Pokemon is superior.
A new, powerful and evil Pokemon trainer could be the antagonist. This trainer might be jealous of Ash's new form and powers as a Pokemon and wants to capture him to use his powers for his own gain. He could set up various traps and battles to try and subdue Ash.
It could be a group of Team Rocket - like villains. They are after the unique powers that the reborn pokemon might possess. Since the reborn pokemon has a human - like intelligence, they see it as a valuable asset to use for their evil plans, like stealing all the precious pokemon resources or taking over a certain region.
It could be a jealous rival trainer. This trainer might have always been envious of Ash's popularity and the loyalty of his harem. So, they plot and scheme to turn the harem against Ash by spreading lies and creating discord among them.
It could be a rival trainer who is jealous of Ash's success and also wants to use Princess Sara for her influence. He spreads false rumors about Ash to turn the people of the princess's kingdom against him. He then tries to take over the role of the hero in the situation, but his real intentions are to gain power and wealth by any means necessary.