This paper presents how, in a calibration process, different assumptions regarding the standard Mixed-Culture Biofilms (MCB) model were able to match the average results at a continuous Johannesburg pilot plant (comprising two aerobic reactors, AE1 and AE2), but failed to match the batch test results of either the rate of endogenous carbonaceous oxygen uptake (OUR) or the rate of nitrate production (NPR). Under the first assumption, where attachment and diffusion of particulate components were not used, the OUR in the biofilm of the first aerobic reactor (AE1) was too low due to the absence of slowly biodegradable COD (XS) attachment flux. In a second assumption, where high diffusion and attachment coefficients were used, the NPR in the biofilm of the AE1 reactor exceeded the experimental value due to the high attachment flux used for nitrifiers (XA) and the low competition for space from XS and heterotrophic bacteria (XH). The only way to match all the experimental results was through the use of a higher attachment coefficient for XS in the first reactor (AE1), but this was considered unreasonable. Hence, an extended model was developed where a colloidal state, which interacts at the same time with the flocs and the biofilm through attachment-detachment processes, is distinguished. This model allowed the experimental results to be matched, but using the same value for the attachment coefficients of all particulate components.

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