It is a dubious but common practice to use growth parameters measured in suspended cultures to predict substrate concentration profiles in biofilms. To obtain biofilm biokinetic parameters that apply to biofilms, a reliable method is needed that allows the computation of biokinetic parameters from substrate concentration profiles measured directly in biofilms. We have developed such a method and demonstrated its utility by evaluating biokinetic parameters from oxygen concentration profiles measured in biofilms of Leptothrix discophora SP-6 grown on a membrane, which was placed on top of an agar plate by fitting the data to Monod or Tessier growth kinetics, including maintenance substrate consumptions. We found that the Monod model represented the growth of L. discophora SP-6 biofilms marginally better than the Tessier model. The Monod half saturation coefficient was 0.333 +/- 0.077 mg/l. (C) 2003 Elsevier Ltd. All rights reserved.