This paper presents the anaerobic biomass characterization and the bacterial framework inside polyurethane foam matrices taken from a horizontal-flow anaerobic immobilized-sludge (HAIS) reactor treating a glucose-based substrate. Ultrastructure polyurethane foam analyses carried out using scanning electron microscopy (SEM) in samples treated with hexamethyldisylazane showed three different patterns of biomass retention inside the polyurethane foam matrices : micro-granules ranging from 270 mu m to 470 mu m were entrapped in the porous medium thin multi-cellular films were attached to the inner surface, and individual cells adhered to the support. The use of SEM and epifluorescence microscopy permitted inferences to be made on the bacteriological composition of the immobilized sludge formed by different morphotypes (rods, cocci and filaments) and on the ecological significance of their framework inside the matrices. Polyurethane matrices were found to offer excellent conditions for anaerobic growth and retention. favoring the flux of substrate and products. Such outstanding characteristics were confirmed by the short start-up period observed during the operation of the HAIS reactor.