The next day, The group Murdoch hires is attacked by the anaconda and it kills one of them.

Due to the serum, the anaconda can no longer die without significant damage to its internal organs and regenerates itself from the previous cavein. Meanwhile, a group of explorers: Jackson, girlfriend Wendy, Wendy's best friend Heather, and brothers Patrick and Scott, get dragged into the search for the snake. Amanda is able to trick the snake into causing a cave in that 'kills' the snake however as she and Alex attempt to leave the mine, she is knocked unconscious and Alex escapes to get more help. When they discover the blood orchids in the mine, Amanda sets the explosives to destroy the orchids, but she and the group are attacked by the anaconda that kills one of the officers with her while the other officer hides and disappears. On the way, they meet Alex, a trekker who appears lost in the Romanian Carpathians, while doing a paleopathology project. Amanda Hayes, the sole survivor of the previous snake attack, and two officers also go in search of Peter, determined to destroy the serum and kill the snake. Peter 'J.D.' Murdoch, a billionaire suffering from bone cancer, sends his assassin Eugene and his team of henchmen mercenaries to find Peter and the serum Peter created so that it can cure him. When he disappears, the long anaconda escapes from the cage and kills Peter in a mine filled with blood orchids. After the test seemingly works for the baby anaconda to regenerate, Peter burns it. A baby anaconda, which was captured at the end of the previous film, is being used for experiment by Peter Reysner, who creates a hybrid of the blood orchids from Borneo that enabled the anaconda to grow very large and live longer, and creates a serum for cell regeneration.

Cast of anaconda