Animal rights campaigners PETA claim they witnessed "cruel" treatment of owls at the Harry Potter Warner Bros Studios tour in Hertfordshire.
A video filmed by PETA shows the animals in cages and being touched by visitors at the popular tourist attraction.
The tour allows Harry Potter fans to meet Harry's owl, Hermione's cat and other animals from the films.
Mimi Bekhechi, director of PETA, told the Sunday People: "Confining frightened owls to tiny cages where they can only chew at their tethers in frustration goes against every message of respect and kindness that JK Rowling's wonderful books taught us.
"PETA are calling on Warner Bros Studio Tour London to make sure that the Harry Potter tour stays magical – and not cruel – by keeping live animals out of it."
Dr Manilal Valliyate, director of veterinary affairs at PETA India, told the Daily Mail: "Owls are wild, solitary, nocturnal predators, and it is completely against their nature to be tethered inside tiny cages, exposed to crowds, flash photography, loud noises and forced to perform 'tricks'."
But Warner Bros Studio Tour London told the BBC: "It is essential the welfare of the birds... is of the highest standard."
It added: "They appear for short periods and are exclusively handled by the experts at Birds and Animals, the company that owns and trains them. We have asked them to review this matter."
A spokeswoman for Birds and Animals told the Sunday People: "The owls are always given regular breaks and closely monitored by a vet. Now that we have had the opportunity to see the footage, we have instigated a review of the issues raised."