Several dairy cows have been identified as partially deficient in UMP synthase. Although erythrocytes ofnormal cows contained 2.54 units of enzyme per ml, four cows were discovered with only 1.08 units per ml. Cows deficient in UMP synthase secreted milk with abnormally high levels of orotate, 300-1,000 pug oforotate per ml compared to 80 ,ug/ml for normal cows. The deficiency also was accompanied by a lactation-induced orotic aciduria. Although bovine urinary orotate was generally <10 #g/ ml, the urine of the deficient cows, when lactating, contained 20-200 #g/ml. Their plasma orotate also was elevated. Genetic transmission of the condition was suggested by a common bull in the pedigrees of all deficient animals. Indeed, these cows, with half the normal level of UMP synthase, are probably heterozygotes with a 50% chance of passing the deficient allele to their progeny. For these putative heterozygotes, the condition is apparently benign because longevity and production were unaffected. However, the existence of a gene for UMP synthase deficiency in the dairy cow population poses a hazard with respect to the conception of homozygotic, deficient animals. These, in analogy with a comparable human condition, would be expected to exhibit high perinatal morbidity and mortality.In mammalian cells, the last step ofpyrimidine nucleotide synthesis involves the conversion of orotate to UMP and is catalyzed by UMP synthase (1). This multifunctional enzyme is comprised of two sequential activities, orotate phosphoribosyltransferase [orotate + phosphoribosylpyrophosphate -* orotidine 5'-monophosphate (OMP)] and OMP decarboxylase (OMP -_ UMP + CO2). A severe deficiency of this enzyme results in hereditary orotic aciduria, a rare disorder in humans. In addition to urinary excretion of massive quantities of orotate (1,500 mg/day vs. 1.4 mg/day in normal subjects), it is characterized by megaloblastic anemia and growth retardation (2). The latter conditions result from de facto starvation for pyrimidines and hence inadequate nucleic acid metabolism. Unless treated with exogenously supplied pyrimidines (usually uridine), afflicted individuals die in the neonatal period.Orotate is a normal constituent of bovine milk and is virtually the only pyrimidine or purine in the acid-soluble fraction (3). Orotate concentrations in milk exhibit considerable variability; although some variation is due to breed, stage of lactation, and number of lactations, it is attributable in large measure to individual variation (4). In a multibreed dairy herd with 250 milking cows monitored for 5 yr, several cows were identified with persistently high levels ofmilk orotate. The herd mean (+ SEM) was 81.1 ± 15.8 p.g oforotate per ml, yet milk from certain cows had >300 Ag/ml and, in one case, it reached nearly 1,000 ,tg/ ml.The purpose of this study was to assess an enzymatic basis for the elevated secretion of orotate by these cows. It was hypothesized that a deficiency of UMP synthase would lead to excess accumulation oforotate in milk. The eryt...