The muskrat (Ondatra zibethicus) is presumed to have undergone a rapid phyletic size decrease near the end of the Pleistocene. Evolutionary changes in the size of middle to late Wisconsinan (ca. 32,000–12,300 14C yr B.P.) muskrats from the Aucilla River, Jefferson County, Florida, were reconstructed by examining length and width of the lower first molar (m1). Body mass, estimated from m1 length, was relatively stable from 32,000 to 16,000 14C yr B.P. and decreased only slightly by 12,300 14C yr B.P. If the size trend found in the Aucilla River material is characteristic of the southeastern United States, a body size decrease after 12,300 14C yr B.P. is needed to explain the smaller size of modern populations. It was previously thought that the length/width (l/w) ratio of the muskrat m1 was a paleoenvironmental indicator based on its presumed correlation with latitude in modern populations. We examined the length and width of modern muskrats from several geographic regions and found only a very weak trend in the size of the m1 between northern and southern populations; however, highly significant differences were found between regions of similar latitude. Our data indicate that chronoclines in the m1 of the Aucilla muskrat material and other such documented trends among fossil muskrats have paleoenvironmental significance, but it is not yet clear which environmental variables can best be predicted from them.