The effect of the substrate/microorganism ratio during the development of anaerobic activity test was studied. The experimentation was carried out in serum bottles at 35°C. Two sets of experiments utilizing acetate and an azo dye (blue disperse 79) as the sole source of carbon were studied. It was observed that mixing has an important influence on the results. The initial substrate concentration and the initial biomass concentration had a significant effect on the reaction rate and on the biomass yield coefficient, Yobs. Different kinetic coefficients were found for the case of equal So/Xo ratio, but different initial substrate concentration.

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