Organic matter quality, expressed as the proportion of chlorophyll a (Chl a) to degraded organic material (i.e. phaeopigments), is known to influence the structure of benthic associations and plays an important role in the functioning of the ecosystem. This study investigates the vertical distribution of microbial biomass, meiofauna and macrofauna with respect to organic matter variation in Ubatuba, Brazil, a southeastern, subtropical coastal area. On three occasions, samples were collected in exposed and sheltered stations, at high and low hydrodynamic conditions. We hypothesize that benthic assemblages will have high meio- and macrofaunal densities and high microbial biomass at the sediment surface at the sheltered site, and lower and vertically homogeneous microbial biomass and densities of meio- and macrofauna are expected at the exposed site. The accumulation of fresh organic matter at the sediment surface was observed at both stations over the three sampling dates, which contributed to the higher densities of meiofauna in the first layers of the sediment column. Macrofauna followed the same trend only at the exposed station, but changes in the number of species, biodiversity and feeding groups were registered for both stations. Microbial biomass increased at the sheltered station over the three sampling dates, whereas at the exposed station, microbial biomass was nearly constant. Physical exposure did not influence organic matter loading at the sites and therefore did not affect overall structure of benthic assemblages, which negates our original hypothesis. Most of the benthic system components reacted to organic matter quality and quantity, but relationships between different-sized organisms (i.e. competition and/or predation) may explain the unchanged microbial profiles at the exposed site and homogeneous vertical distribution of macrofauna at the sheltered site. In conclusion, the high quality of organic matter was a crucial factor in sustaining and regulating the benthic system, but coupled results showed that interactions between micro-, meio- and macrofauna can be highly complex.