The concentrations of holotrichs, reducing sugars and fermentation end-products, and the fluid dilution rates, in the cranial (CR), ventral (V) and caudal (CA) regions of the reticulo-rumen of cows were compared. Additionally, holotrich numbers in fluid and solid digesta samples taken from the dorsal (D) region were determined at different time-intervals. Holotrich numbers expressed per ml fluid at site D were almost twofold those expressed per g solid digesta, and the fluctuation of their numbers in fluid preceded that in solid digesta. Holotrich concentrations at site CR were higher before feeding than after feeding, while those at sites V and CA were lower before feeding than for a few hours after feeding. At sites V and CA, holotrich concentrations fluctuated in good agreement with reducing sugar concentrations, but the increase in the former always preceded that in the latter. The concentration of volatile fatty acids (VFA) tended to decrease from site CA to site CR, while the dilution rate was highest in site CR. The results suggest that migration of holotrichs from the reticulum to the rumen and vice versa are the cause of fluctuation in their numbers, and also the mechanism by which they are retained in the rumen of cattle.