Saizo had a Barbaracle, who tested Sanpei's skills as a ninja and a trainer. Barbaracle managed to poison Sanpei's Greninja, who was later healed by the heroes. Later, Saizo sent Barbaracle to attack Sanpei and Greninja once more, but Team Rocket interfered, so Barbaracle blasted them off. Once done, it proceeded to attack Greninja, but Ash's Froakie evolved into Frogadier and protected Greninja. Saizo was satisfied and disappeared with his Barbaracle.
Barbaracle protected Ash's Pikachu, Ash's Frogadier and Sanpei's Greninja from Heidayu's Bisharp's Snarl when Kagetomo's army attacked the Ninja Village. Barbaracle then used Razor Shell on Bisharp, who was defeated by that attack.