The present study was undertaken to examine whether pregnancy diagnosis was possible by measuring fecal progesterone concentrations in beef and dairy heifers and beef cows. Rectal fecal samples collected on days 18-24 after insemination or days 11-17 after embryo transfer were mixed with methanol and shaken for preparation of a fecal solution. After centrifugation, the supernatant was extracted with petroleum ether followed by an enzyme immunoassay for progesterone. All pregnant animals showed fecal progesterone concentrations greater than 50 ng/g of fecal material on days 18-24 after AI or estrus. In non-pregnant animals, however, the fecal progesterone concentrations ranged widely from 5 to 180 ng/g of fecal material. In non-pregnant cattle, the percentage of cattle with <50 ng progesterone/g of fecal material compared with the total number was 37-60% on days 18-20, whereas the percentages increased more than 70% to a maximum of 78.1% on day 23. When 50 ng/g was considered as the cut-off value, the sensitivity and specificity of positive pregnancy tests were less than 70% on days 21-24, and 100% for negative pregnancy tests on days 18-24. There were significant differences in the mean fecal progesterone concentrations between pregnant and non-pregnant cattle on days 19-24. These results suggest that feces can be utilized to substitute for plasma and milk to measure progesterone for the purpose of pregnancy diagnosis in heifers and cows.