The purpose of this retrospective study was to evaluate the repositioning accuracy of different positioning devices in order to determine their applicability for potential use in conformal radiation therapy for animals. Forty-four animals with spontaneous tumors of the head were included. The animals were divided into 3 groups determined according to the positioning device used. Group 1 animals were positioned using a thermoplastic mask. Group 2 animals were positioned using a head holder. Group 3 animals were positioned using the head holder and an inflatable pillow. The time of presentation determined which position device was used. Port films of the 44 patients were reviewed retrospectively, and the repositioning precision was recorded by measurements in three orthogonal planes. Groups 2 and 3 had significantly better repositioning accuracy (P < 0.05) compared to Group 1. The position variation was not significantly different (P < 0.05) between Groups 2 and 3 in the lateral and longitudinal direction. Group 3 had a median reposition variation of 0.5 to 1.0 mm, with a standard deviation of 1.0 to 1.5 mm.