The moisture content of horse hoof horn is important as it affects its function, quality and mechanical properties. It is believed for horse hoof horn that there is an inverse relationship between the amount of water present and stiffness, or the ability of hoof horn to resist deformation. The majority of studies, until the work of Leach (1980) and Douglas et al. (1996) concentrated on quantitative values for moisture content for the full hoof wall depth (HWD) for horse hoof horn. Douglas et al. (1996) divided the hoof wall into inner and outer wall samples and showed a moisture content of 28% for the outer wall and 35% for the inner wall, confirming a dorsopalmar increase in moisture content. Whether a similar gradient exists for donkey hoof horn is not known. The existence of such a dorso-palmar moisture gradient may have a profound effect on the mechanical properties of the hoof horn and would further contribute towards the understanding of how moisture content affects the function of the hoof.