A transient mathematical model was established in order to evaluate oxygen diffusivity in non-steady-state biofilms. A submerged fixed bed biofilm system with efficient medium recirculation was investigated for p-toluenesulphonic acid degradation by Comamonas testosteroni T-2 in a multispecies biofilm. Static mixer elements (Sulzer Chemtech Ltd, Switzerland) were used as a support matrix for biofilm formation. Biofilm respiration was estimated using the dynamic gassing-out oxygen uptake method. Based on the dissolved oxygen concentration profiles, the oxygen diffusion coefficient and the maximum respiration activity were calculated. The values of the dissolved oxygen diffusion coefficient varied with biolfilm development and values reported here (2 x 10(-10)-1.2 x 10(-9) m(2) s(-1))are in good agreement with literature data. Calculated oxygen consumption rates fit well with values obtained in respirometry tests with washed out biofilms. The knowledge of diffusivity changes in biofilms is particularly important for removal capacity estimation and appropriate reactor design.