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The moisture content of keratinous materials such as hoof horn is important as the presence of moisture has an inverse relationship on the mechanical properties of hoof horn and may have a subsequent effect on the function of the hoof. Methods previously used to dehydrate samples to calculate the moisture content of hoof horn vary considerably (Hopegood, 2002). Subsequent comparison of results is therefore unreliable. A comparison of different methods of dehydrating hoof horn was therefore carried out to establish a standardised protocol for dehydrating hoof horn to assess its moisture content. The moisture content of donkey hoof horn from normal animals and those with laminitis has not been reported. Maclean (1971) established that the moisture content of cattle suffering from laminitis was significantly higher than normal hooves. The resultant standardised protocol from the first part of this study was then used to compare the moisture content of hoof horn samples taken from horses, donkeys and those donkeys that had suffered from laminitis.