A study was conducted to obtain data on the effects of a fungal fibrolytic enzyme preparation (Rumino-zyme, with 250 FXU/g xylanase activities) from Thermomyces lanuginosus on some rumen fermentation parameters in sheep. Ruminal fluid samples were taken just before the morning feeding and then 2 h and 4 h after feeding. Xylanase activity, pH, concentration of ammonia and volatile fatty acids were measured. The enzyme supplementation did not affect the pH but increased the xylanase activity and the total VFA concentration of the rumen fluid. The molar proportion of acetate increased, propionate was not affected and butyrate decreased after enzyme administration. The concentration of ammonia also decreased after supplementation with the enzyme product. It can be concluded that the xylanase enzyme preparation from T. lanuginosus induced favourable changes in the major rumen fermentation parameters in sheep.