“…Research in the last 40 years has completely changed the traditional understanding of substrate and biomass and has paved the way for the evolution of activated sludge modelling that enables numerical interpretation of the major biochemical mechanisms taking place in the reactor. Key achievements in the area should be noted as follows: (i) Adoption of COD as the sole parameter for organic substrate and biomass -This way, an electron balance could be established between COD utilized, biomass generated, and oxygen consumed, 28,29 (ii) Identification of COD fractions with different biodegradation characteristics; similarly, 23 differentiation of biomass into fractions with different functions, 30,31 (iii) development of multi-component biokinetic models including all substrate and biomass fractions as model components, 32 and (iv) Experimental support of respirometric techniquesthis support enabled the determination of process kinetic and stoichiometry by model calibration of oxygen uptake rate profiles. 33 Process stoichiometry is a useful instrument for assessing the major characteristics of the activated sludge process as a function of two major variables selected for system operation, namely, the sludge age, ⊔ X , and the hydraulic retention time, ⊔ H .…”