Abstract.Fescue toxicosis research studies have often included serum prolactin as a physiologic index of the disorder. Serum prolactin has not been used as a clinical measure of fescue toxicosis because of variation associated with sex and physiologic condition of the animal and climatic and seasonal factors. The primary excretory route of the alkaloids responsible for this toxicosis is the urine. Three pasture experiments were conducted to examine serum prolactin and urinary ergot alkaloid variability among steers continuously grazing endophyte-infected (Eϩ) or endophyte-free (EϪ) tall fescue and among steers that were switched from one pasture form to the other. A fourth grazing experiment was used to examine how to best to manage the steers prior to sampling for urinary ergot alkaloid excretion. Coefficients of variability for urinary alkaloid excretion were consistently lower (46-65%) than serum prolactin (64-142%). Urinary alkaloid excretion patterns changed within 12 hours following switching steers from Eϩ to EϪ pasture or visa versa, but serum prolactin was recalcitrant to change. Because it is less variable and more dynamic than serum prolactin, urinary alkaloid excretion can be used for health assessment of steers grazing Eϩ and EϪ pastures. Regression analysis established a quadratic relationship between alkaloid excretion and average daily weight gain, with a regression coefficient of 0.86. Urinary alkaloid analysis was useful in determining whether cattle were consuming toxic tall fescue.Diagnostic tools used to determine potential for toxicity or toxic effects of endophytic fungus-infected tall fescue include plant (pasture or hay) sampling for ergot alkaloids, sampling the plant for endophyte, or sampling serum from afflicted animals for suppressed prolactin, alkaline phosphatase, or cholesterol. There has been no established cause/effect relationship between serum alkaline phosphatase or serum cholesterol and fescue toxicosis; hence, their use as physiologic markers may be compromised for the disorder. 3,8,11,19 Prolactin is highly variable depending on animal gender, 15 reproductive state, 1 day length, 10 season of the year, 14 and temperature. 21 Therefore, serum prolactin, cholesterol, and alkaline phosphatase have liabilities as markers for toxicosis. However, in research studies with proper controls, serum prolactin is the most commonly analyzed physiologic marker used to confirm physiologic effects of fescue toxicosis. 4,5,9,12,16 Analysis of plant tissue for alkaloids or endophyte is easily performed, 6,7 but the assumption that animals are consuming the toxic forage may be incorrect if pastures are not highly infected or if other plant species are present in the pasture. Therefore, a direct diagnostic tool is needed to determine if ergot alkaloids in pasture plants Received for publication June 17, 1999. are in fact ingested by the animal and whether ergot alkaloids can be used as an index of fescue toxicosis. In a recent study, 17 urinary and biliary excretion of ergot alkaloids from...