This work focused on a morphological study of the microorganisms attached to polyurethane foam matrices in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor treating domestic sewage. The experiments consisted of monitoring the biomass colonization process of foam matrices in terms of the amount of retained biomass and the morphological characteristics of the cells attached to the support during the start-up period. Non-fluorescent rods and cocci were found to predominate in the process of attachment to the polyurethane foam surface. From the 10th week of operation onwards, an increase was observed in the morphological diversity, mainly due to rods, cocci, and Methanosaeta-like archaeal cells. Hydrodynamic problems, such as bed clogging and channeling occurred in the fixed-bed reactor, mainly due to the production of extracellular polymeric substances and their accumulation in the interstices of the bed causing a gradual deterioration of its performance, which eventually led to the system's collapse. These results demonstrated the importance and usefulness of monitoring the dynamics of the formation of biofilm during the start-up period of HAIB reactors, since it allowed the identification of operational problems.
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