Cows’ milk containing elevated concentrations of Se provides a rich nutritional source of this essential element for meeting daily nutritional requirements or providing health benefits in humans with low immune function or at risk of cancer. An experiment involving either 2 or 6 wk of dietary supplementation with Se yeast (with the yeast supplying about 30, 40, and 60mg of Se/d for cows supplemented for 2 wk, and about 20, 30, 40, and 60mg of Se/d for cows supplemented for 6 wk), and 21 wk of monitoring of Se status after the withdrawal of supplementation, was undertaken between September 2008 and April 2009 using 35 multiparous Holstein-Friesian cows. Using milk and blood Se concentrations as surrogates, the research examined the time taken for Se build-up in tissue due to supplementation of lactating dairy cows with Se yeast to dissipate back to normal levels. At the end of Se supplementation, a significant relationship was found between milk Se concentration and Se intake, whereby milk Se concentration had increased by 4.5μg of Se/kg of milk for each mg of Se eaten per day, but no effect of duration of supplementation on this relationship was observed. At the same time, both Se intake and duration of supplementation affected blood Se concentration; it increased by 3.6μg of Se/kg of blood for each mg of Se eaten per day, and was 86μg of Se/kg higher after 6 wk compared with 2 wk of supplementation. After the withdrawal of Se supplementation, milk Se concentrations responded quickly to the change in the quantity of Se consumed, and again, duration of supplementation had no effect on the response, but any effect that Se intake had on milk Se had completely dissipated by 4 wk. In contrast to milk, blood Se concentrations continued to be affected by both amount and duration of Se supplementation for at least 4 mo after the withdrawal of supplementation, although by 5 mo the effects of the previous supplementation treatments had virtually disappeared. The slow decline in blood Se concentrations after the withdrawal of supplementation would most likely be due to the protracted clearance of Se from the various tissues that had accumulated Se during supplementation and the rate of erythrocyte turnover. When undertaking an on-farm Se enhancement program to generate milk for the manufacture of Se-enriched milk products, post-supplementation milk Se concentrations are unlikely to create any problems at the milk factory beyond 4 wk, but the high residual blood/tissue Se concentrations that take considerably more time to dissipate may provide the potential for possible unintended consequences at the food chain/farm environment level.