Serum activities of gamma-glutamyl transferase (GGT), alkaline phosphatase (AP), aspartate aminotransferase (AST), and concentrations of total bilirubin and total bile acids were screened during a 5 year period in 27 horses used for production of hyperimmune serum. The horses investigated were regularly immunized with live cultures of the endotoxin-releasing bacteria Escherichia coli or Pasteurella multocida, the individual animals having undergone such treatment for periods varying from 2 weeks to 10 years. In a majority of the horses, GGT-activity had increased within 6 to 7 years of first having undergone immunization. Constantly high values seemed to co-incide with the presence of advanced liver amyloidosis, as demonstrated by histopathological examination after slaughter. The AP activity was also increased but only moderately compared with GGT. Individual values more than 10-fold greater than the upper reference limit were recorded for GGT, while the highest AP values were less than double the upper reference limit. Activity of AST and bilirubin concentrations remained unaffected, whereas the concentration of total bile acids rose after 6 to 7 years of immunization compared to the baseline value. It is concluded that the determination of serum activities of GGT may serve as a practical routine test for the evaluation of liver amyloidosis status in serum horses.