This paper represents the second part of a study searching for factors which could be responsible for an inferior hoof horn quality of the Lipizzaner horses of the Viennese Spanish Riding School (SRS) noticed in the late 1980s. It includes an evaluation of a treatment with biotin and 42 Lipizzaner horses were tested in a double blind study. The following parameters were evaluated: 1) the histology of the hoof horn in samples from the bearing border 2) the tensile strength of the coronary horn in bearing border samples and 3) the influence of biotin upon the histological changes and the tensile strength after application of 20 mg/day of biotin during 38 months. The histological alterations of bearing border specimens were assessed by use of a grading system (Grade 0 = unchanged; Grade 1 = slight changes; Grade 2 = moderate changes; Grade 3 = severe changes). Initially more than two thirds of the horses showed moderate to severe changes: microcracks visible in the transition from the middle to the inner zone of the coronary horn; separation of the sole from the coronary horn in the region within the white zone. The tensile strength of the coronary horn, with a mean of 39 N/mm2, was 13 N/mm2 lower compared with the unchanged hooves of 10 Warmblood horses, which were investigated by Küng (1991). After 19 months of biotin treatment, the horn quality showed a small but significant improvement. This could be shown by comparing the histological tissue structure before and after treatment, as well as treated animals vs. a placebo group.(ABSTRACT TRUNCATED AT 250 WORDS)