Microbial biomass development in the sand and schmutzdecke layer was determined in two full-scale slow sand filters, operated with and without a light excluding cover. A standard chloroform fumigation-extraction technique was adapted to routinely measure microbial biomass concentrations in the sand beds. Sand was sampled to a depth of 10 cm and schmutzdecke was also collected at the same random positions on the uncovered filter. Interstitial microbial biomass in the uncovered sand bed increased with time and decreased with sampling depth. There was a small accumulation of sand biomass with time in the covered filter, but no relationship was apparent between biomass concentration and depth in this filter. Schmutzdecke did not develop on the covered filter and was spatially highly variable in the uncovered condition compared to the consistent patterns observed in interstitial biomass production. It is speculated that microbial biomass in the sand of uncovered filters is largely related to carbon inputs from photosynthetic activity in the schmutzdecke and involves mechanisms that spatially distribute carbon substrate from the schmutzdecke to the sand. However, total organic carbon and dissolved organic carbon removals were similar in both filters suggesting that relatively small biomass populations in covered filters are sufficient to remove residual labile carbon during advanced water treatment and little further advantage to water purification and organic carbon removal is gained by the increased production of biomass in uncovered slow sand filter beds.