Studies were conducted to investigate the effects of lead and niacin on tryptophan and serotonin metabolism in growing broiler chicks. A low-niacin basal diet based on soybean meal and glucose was supplemented with either 40 ppm niacin/kg feed or 2,000 ppm lead, as lead acetate trihydrate, or a combination of the two in a 2 x 2 factorial arrangement. The experimental diets were fed from 1 day to 3 weeks of age. The activities of several enzymes involved in tryptophan and serotonin metabolism were assayed in chicks fed each of the experimental diets. Lead reduced the activity of liver tryptophan pyrrolase but had no effect on the activity of hepatic picolinic carboxylase. Low dietary niacin had no effect on the activity of either of these enzymes. Brainstem tryptophan 5-hydroxylase activity was unaltered by either lead or niacin. Brain and liver monoamine oxidase activity was reduced by lead but was not affected by niacin. No interactions of lead and niacin were observed with any of the enzymes examined. Lead had no consistent effect on brain serotonin (5-HT) steady-state level, but it increased the level of the major metabolite of 5-HT, 5-hydroxyindoleacetic acid (5-HIAA). Monoamine oxidase blockade failed to reduce the elevated 5-HIAA level in lead-treated chicks. The drug probenecid produced an increase in 5-HIAA that was comparable to the increase caused by lead. The effects of probenecid and lead were additive. It is concluded that lead significantly altered the activity of several enzymes involved with tryptophan metabolism, whereas the dietary niacin levels employed were without effect. Additionally, lead caused the accumulation of 5-HIAA in the brain, which appeared to result from inhibition of the probenecid-sensitive acid transport system.