Earlier weaning has often been suggested as a cause for population growth after the evolution of food production. However, evidence for weaning-time reduction is largely circumstantial. Collagen stable nitrogen- and carbon-isotope ratios from juvenile and adult burials from four sites in eastern North America were measured to estimate weaning onsets and durations before and after the appearance of intensive food production. Two preagricultural Late Archaic sites (Indian Knoll and Carlston Annis) are compared with two highly agricultural Middle Mississippian sites (Angel and Tinsley Hill). Isotopic data and paleodemographic measures of birth rates provide no evidence for changes in weaning behavior or fertility with the development of food production in the prehistoric Lower Ohio Valley. Birth rates and weaning behavior appear to have been roughly the same at all four sites. These results indicate that models attributing population growth after the appearance of food production to earlier weaning are not universally applicable. Am J Phys Anthropol, 2004. © 2004 Wiley-Liss, Inc.