OBJECTIVE To determine the optimal protocol for acquisition of CT images of the dentition in alpacas. ANIMALS 3 healthy adult male alpacas. PROCEDURES Each alpaca was anesthetized with an IM injection of a combination of ketamine, xylazine, and butorphanol and positioned in sternal recumbency on the CT couch with its legs folded in a natural cush position and its head positioned within the isocenter of the gantry of a 64-slice CT scanner. Images were acquired by means of 6 protocols (sequential and helical modes at slice thicknesses of 1.25, 2.5, and 5 mm). Five images (2 molar, 2 premolar, and mandibular incisor teeth) were selected from each protocol for evaluation by 3 veterinary radiologists. For each image, tooth root visibility and sharpness and image noise artifact were subjectively evaluated on a 3-point scoring system. RESULTS Slice thickness significantly affected tooth root visibility and tooth root sharpness but did not affect image noise artifact. Acquisition mode significantly affected tooth root visibility and tooth root sharpness as well as image noise artifact. Tooth root visibility and sharpness did not differ significantly between the helical and sequential images when the slice thickness was 1.25 mm. Image noise artifact was greater for helical images than sequential images but did not differ by slice thickness within either acquisition mode. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that for a 64-slice CT scanner, the optimal protocol for the acquisition of CT images of the dentition in alpacas was a sequential scan with a slice thickness of 1.25 mm.