Four vitamin B12 assays were compared using blood sera from Friesian cows on winter diets or grazing. In herd 1, ten animals were blood-sampled three times at monthly intervals and the vitamin B12 concentration of the sera determined by the Poteriochromonas malhamensis and Lactobacillus delbrueckii assays. At all three sampling dates the results produced by the P. malhamensis assay were significantly greater than those produced by the L. delbrueckii assay. Cows in herd 2 were divided into two groups, each of 30 animals. One group was given a soluble glass bolus releasing cobalt and the other was unsupplemented. Milk yields were recorded throughout the experimental period and herbage samples were taken at intervals for cobalt determinations. Blood samples were taken at intervals from January to September 1983 and the vitamin B12 concentration of the sera determined by four different assay methods: the P. malhamensis and L. delbrueckii microbiological assays and the Becton Dickinson and RIA Products ‘No-boil’ radioassays. The last of these failed to detect vitamin B12 in any sample. There was a significant difference between the results obtained by the three other assays, with the Becton Dickinson radioassay consistently producing the lowest result and the P. malhamensis assay the highest result of the three. There was no significant effect of cobalt supplementation on milk yield.