The effect of ammonia treatment of straw on both the rumen environment and the extent of its microbial fermentation was studied. Four rumen cannulated sheep were randomly given 700 g/day of untreated straw plus urea (US), ammonia-treated straw (TS) and alfalfa hay (AH) in a change-over design with three periods. Rumen pH was lower and ammonia-nitrogen and total volatile fatty acid (VFA) concentrations were higher (P < 0·001) with AH than with US or TS. With the straw diets, TS promoted a lower pH than US (P < 0·05), but differences were less than 0·3 units and the mean pH was never below 6·5. There were no differences between the straw diets in ammonia-nitrogen or VFA concentration (P > 0·05). When untreated barley straw (BS) and treated straw (TS) were incubated in situ disappearance of dry matter (dDM) at 12, 24 and 48 h (P < 0·01) and neutral-detergent fibre (dNDF) at 48 h (P < 0·001) were higher with TS. In vitro incubation showed a higher gas production with TS only after 36 h (P < 0·05) whereas gas from BS fermentation was higher up to 14 h (P < 0·05). Among diets, dDM, dNDF and gas production with US were numerically higher than with TS or AH throughout, although few significant differences were observed, except for a higher dDM at 12 (P < 0·01) and 24 (P < 0·10) h and a higher dNDF at 12 h (P < 0·10). Particle-associated enzymes were extracted from BS and TS incubated in the rumen for 4, 8, 22 and 24 h. Results ofxylanase and cellulase activities support those of straw incubation, with a drop between 4 and 8 h in TS diet. The concentration of residual phenolics per unit of incubated straws after 12 and 24 h show that phenolics release to the media was higher with the TS diet. Daily changes of phenolic concentration into rumen liquid was also higher with TS than with US (P < 0·001). The increased release of straw phenolics by ammoniation reduced the potential for rumen degradation of straw, mainly in the first hours of the fermentation period.