Sixteen male Suffolk lambs fed a 8 ppm Cu basal diet were randomly assigned to 2 groups: 12 copper-loaded (CL) and 4 controls (C). The CL sheep were drenched initially with 3 mg Cu/kg bw daily for a week. Every week an additional dose of 3 mg Cu/kg bw was included in the drench until signs of copper poisoning appeared; the control sheep were drenched with saline solution. The onset of copper poisoning occurred between 42 and 55 d. Food intake and body weight were recorded daily. Blood samples were collected weekly to measure the activity of the liver enzymes gamma-glutamyltransferase (gammaGT), aspartate aminotransferase (AST), sorbitoll dehydrogenase (SDH) and acid phosphatase (AF). The following changes were significantly recorded in the CL sheep in the weeks or days previous to the hemolytic crisis: higher levels of gammaGT were found on the -28th d increasing slowly but continuously until the hemolytic crisis; SDH fluctuated during the period presenting higher levels on the -28th, -14th and -7th d; AST and AF activities increased from the -14th and -7th d respectively; sharp decreases in the activities of SDH and AF at the hemolytic crisis; lower feed intake and body weight gain from the -7th d; and sheep ceased eating concentrates from the -9th d and became anoretic the day before the hemolytic crisis. Plasma copper concentration increased only the day before the hemolytic crisis. There were no changes in respiratory and heart rates, rectal temperature or rumen movements throughout the pre-hemolytic phase. The higher the amount of cumulative copper drenched, the higher was the gammaGT and AST activities. It was concluded that gammaGT followed by AST are the best enzymes to assess copper-load in sheep during the pre-hemolytic phase. Sheep fed copper-rich diets with high plasma activity of these enzymes, decreased feed consumption and subtle loss of body weight are most likely to present with a hemolytic crisis in a few days.