The use of live animals in U.S. medical education declined progressively from 1985 to 1994. This study determined whether this trend continued into 2001. The authors used postage-paid postcards and follow-up e-mails, faxes, and telephone calls to survey department chairs of pharmacology, physiology, and surgery in all 125 U.S. medical schools about the use of live animals in their undergraduate medical courses. The response rates were 97% for physiology and surgery departments and 100% for pharmacology departments. Respondents reported that live animals were used in 5% of pharmacology courses and 18% of physiology and surgery courses. When live-animal laboratories were used in the curricula, attendance was optional in 76% of the courses, and the percentages of students opting out of the laboratories varied from none to over half. The most commonly used animals were pigs and dogs, followed by rodents, rabbits, and cats. During the last seven years, the use of live animals in medical school pharmacology and physiology courses has continued to decline, but the use of live animals in surgery courses has remained stable. Overall, the majority of U.S. medical schools (68%) do not use live animals in any of their pharmacology, physiology, or surgery courses, and when live-animal laboratories are used, attendance is usually optional.