This study aimed to elucidate whether sheep can increase ruminal glucose absorption acutely to counteract lactic acid formation after a concentrate meal. Pre-incubation of Ussing-chambered ruminal epithelia with D-glucose (10 mM) on both sides for 180 min increased the apical uptake of 0.2 mM C-14-D-glucose. An equivalent effect was seen after serosal but not mucosal pre-incubation with D-glucose for only 10 min. However, a 30-s serosal pre-incubation with D-glucose did not stimulate D-glucose uptake. Stimulation of uptake by serosal D-glucose was not seen after phlorizin treatment. Thus, ruminal glucose absorption via SGLT-1 can be up-regulated acutely via a basolateral glucosensor.