The relationship between the pulmonary toxicity of 3- methylindole (3MI, skatole) and the mixed-function oxidase (MFO) system was investigated. Nine goats assigned to three groups were given a jugular infusion of [14C]3MI (0.02 to 0.03 g of 3MI/kg of body weight containing 0.5 muCi/kg of body weight) for 1.5 hours to induce acute pulmonary edema. Two groups of three goats each were treated with phenobarbital (PB) or piperonyl butoxide (BT) prior to 3MI infusion to induce or to inhibit the MFO system. Three goats were used as 3MI controls. During a 72-hour test period, blood was collected for determination of plasma 3MI concentration and radioactivity. Urine was collected and was fractionated by column chromatography. The severity of pulmonary lesions was evaluated by gross and microscopic examination. Pretreatment with BT prevented the onset of acute pulmonary edema. Goats pretreated with PB had more severe lung lesions than did 3MI controls. Plasma of goats pretreated with BT had a longer half-life (2.1 hours) of radioactivity, whereas plasma of goats pretreated with PB had a shorter half-life (1.0 hour) when compared with plasma of 3MI control goats (1.5 hours) given the same dosage of [14C]3MI (P less than 0.025). The plasma half-life of 3MI was longer (P less than 0.025) in BT-pretreated goats (0.45 hour) than that in PB-pretreated goats (0.26 hour). At 72 hours, 70% to 98% of the infused radioactivity had been excreted in the urine. The pattern of urinary metabolites of 3MI was altered in BT-pretreated goats compared with patterns in control and PB-pretreated goats. Results indicate that the MFO system is one of the pathways involved in the metabolism of 3MI and that pulmonary toxicosis results from metabolism of 3MI by this enzyme system.