Stems of orchardgrass hay in nylon bags were incubated in the rumens of three ruminally fistulated sheep to monitor the rate and extent of fiber attachment by the representative ruminal cellulolytic bacteria via competitive polymerase chain reaction. After incubation for 5 min, the numbers of Fibrobacter succinogenes and the two ruminococcal species attached to stems were 10(5) and 10(4)/g dry matter (DM) of stem, respectively. At 10 min, the numbers of all three species attached to stems increased 10-fold. Thereafter, attached cell numbers of the three species gradually increased and peaked at 24 h (10(9)/g DM for F. succinogenes and 10(7)/g DM for Ruminococcus flavefaciens) or 48 h (10(6)/g DM for Ruminococcus albus). On the other hand, cell numbers of all three species in the whole digesta were constant over 24 h. Changes in the rate of in sacco neutral detergent fiber disappearance of hay stem, which showed a linear increase up to 96 h, were not synchronized with changes in cellulolytic bacterial mass. These results suggest that sufficient numbers of cells of the three cellulolytic species to move to new plant fragments are present at the start of incubation, the initial attachment to new plant matter is mostly accomplished within 10 min and then bacterial growth and fibrolytic action follow. F. succinogenes was most dominant, both in the whole rumen digesta and on the suspended hay stems, demonstrating the ecological and functional significance of this species in ruminal fiber digestion.