Rumen-cannulated Holstein cows were used to study the effect of intraruminal dosing of dioctyl sodium sulphosuccinate (DSS; 0.07 g/kg body weight per d) for 4 weeks. DSS was suspended in nylon bags to allow it to be released slowly into the rumen. Cows were offered a diet containing grass silage and concentrate (45:55, w/w). Intakes of control cows were regulated to those of DSS-dosed cows. Cows dosed with DSS had no rumen ciliate protozoa, lower rumen NH3-N concentrations and acetate and butyrate proportions, higher propionate, isovalerate, and valerate proportions. In vitro fibre digestion by non-ciliate rumen fluid from DSS-dosed cows was apparently impaired. When cows were dosed with DSS, levels of neutral- and acid-detergent fibre in whole rumen contents were increased, rumen solids turnover rate was slower, and whole tract apparent digestibility of cellulose and diethyl ether extract was decreased. Dosing of DSS led to reduced concentrations of blood acetoacetate but elevated plasma glucose levels. Milk protein content was higher, however, lactose content was lower for DSS-dosed than control cows. Milk fat of DSS-dosed cows had a smaller proportion of short-chain fatty acids but a greater proportion of unsaturated fatty acids.