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An experiment was carried out to study the effect of monensin administration on mammary functions in crossbred Holstein cattle. Fourteen non-pregnant late lactating crossbred Holstein cattle, approximately 270 days postpartum, were selected for the experiment. They were divided into two groups of 7 animals each. Seven animals in the treated group were given sodium monensin orally in a slow-release capsule. Animals in both control and treated groups were fed the similar diet to maintain milk production and body score at 2.5. Rice straw was fed as a source of dietary fiber throughout the experimental period. After monensin administration, a significant increase in the molar percent of ruminal propionate (p<0.05) and a significant decrease in the molar percent of ruminal acetate (p<0.05) were apparent in comparison to the pretreated period. The ratio of acetate to propionate concentration decreased significantly after monensin administration (p<0.05), while it was maintained at the similar level throughout the period of experiment in the control group. Monensin did not affect the molar percent of ruminal butyrate and valerate. The concentration of milk allantoin between the control group and monensin treated group was not different. An excretion rate of allantoin in milk decreased in animals treated with monensin (p<0.05). Mammary blood flow did not show significant difference between control and monensin treated groups. The plasma glucose concentration, arteriovenous concentration difference and mammary gland uptake of glucose remained constant in both groups. Milk yield of the later stage of lactation in the control group declined during lactation advance while a tendency to increase in the milk yield was apparent after 21 days monensin administration. Milk compositions for concentration of lactose, fat and protein in both control group and monensin treated group did not change throughout the experimental periods. From these results, it can be concluded that the action of monensin could affect the ruminal fermentation pattern. Monensin could not increase milk yield in the late lactating period.