The effects of various concentrations of sulfate, organic sulfur, and organic carbon on sulfate uptake by aerobic bacteria were studied using pure cultures growing in a defined medium. Cultures of Pseudomonas fluorescens and Corynebacterium striatum took up sulfate faster when young, but sulfate uptake by Serratia marcescens was faster in older cultures. Organic sulfur was found to decrease sulfate uptake, but at concentrations somewhat higher than occurs in most natural freshwater ecosystems. Low levels of sulfate can theoretically directly limit bacterial biomass production but such limitation probably does not occur in natural systems. Evidence is presented which indirectly links the uptake of sulfate and organic carbon, adding credibility to the proposal that sulfate uptake can be used as an indicator of microbial biomass production in freshwater ecosystems.