Severe fragmentation as a result of the introduction and expansion of mechanized agriculture has decimated the once dominant tallgrass prairie throughout much of the mid-western and mid-southern United States. In Arkansas, the Arkansas Natural Heritage Commission is charged with managing many of the remaining remnant tallgrass prairie fragments as Natural Areas. Though managed nearly annually by prescribed burning, a number of Natural Areas in Arkansas are hayed at least once a year. Since haying removes aboveground vegetation, many essential plant nutrients contained in that vegetation are also removed and are not returned to the same soil from which they were extracted. Therefore, the objective of this study was to evaluate the effects of vegetation removal (i.e., haying) on soil physical, chemical, and biological properties of a native tallgrass prairie fragment in east-central Arkansas. The study site was the Konecny Prairie Natural Area, which contains three areas of differing durations of vegetation removal by haying. Soil bulk density and bacterial biomass concentration in the top 10 cm were highest in the prairie area where vegetation removal by haying is still being conducted, while the prairie area where vegetation removal by haying ceased in 1998 had the highest soil organic matter, but lowest Ca and Mn contents. The effects of duration of vegetation removal by haying on native soil quality were variable and, for some soil properties, unexpected. Remnant prairie fragments can provide benchmark soil data for many properties to help ascertain the effects of land use change, particularly conversion to cultivated agriculture, and can provide target values for soil properties in ecosystem restoration efforts.