Changes of serum transaminse (GOT,GPT) and bilirubin values after a single oral or intraperitoneal administration of α-naphthylisothiocyanate (ANIT) were studied in rats and dogs. After the administration of ANIT, serum transaminase and bilirubin values increased with the increase in dose of ANIT in both rats and dogs, however, the degree of increase in these parameters was much lower in dogs than in rats. The degree of increase in the serum parameters after ANIT treatment in rats or dogs was similar in both oral and intraperitoneal administration. The administration of ANIT produced a marked cholestatic response in rats, but no cholestasis was produced in dogs even when high doses of ANIT were administered or the lapse of time after ANIT administration increased. These differences in response to ANIT between rats and dogs may be explained on the basis of different rates and/or pathways of biotransformation of ANIT.