Studies on the structure of the distal phalanx help explain the development of laminitis. Additionally, examining the structure of the distal phalanx from a taxonomic perspective also contributes to veterinary anatomy. In this study, we examined shape variation in the medial and lateral distal phalanx of both fore- and hindlimbs using the geometric morphometry method. We investigated whether the shape of the distal phalanx differed between phalanx positions and how much of the shape variation in this bone depends on size. For this purpose, distal phalanges from 20 Holstein cattle were used, and the bones were digitized in 3D. A draft containing 176 semi-landmarks was prepared for shape analysis, and this draft was applied to all samples using automated landmarking through point cloud alignment and correspondence analysis. A principal component analysis was performed to obtain general patterns of morphological variation. The centroid size (CS) was employed as an approximation of size. Although distal phalanx groups generally showed close variations, PC1 statistically separated the hindlimb lateral distal phalanx (HL) and the forelimb medial distal phalanx (FM) from each other in shape. While PC2 separated HL from other distal phalanx groups, PC3 separated fore- and hindlimb groups. The shape (Procrustes distance) of the hindlimb medial distal phalanx (HM) is markedly less variable than the other three phalanges. The smallest distal phalanx in size was HL. For both forelimb and hindlimb, the medial distal phalanges were larger than the lateral ones. Size (CS) was found to have an effect on PC1 and PC3. In this study, a reference model of the same breeds for distal phalanx was created. These results can provide useful information, especially in terms of veterinary anatomy, zooarchaeology, and paleontology.