An improved protocol for extracting extracellular polymeric substances from granular filter media

Extracellular polymeric substances (EPS) are an important parameter in drinking-water biofilters, and, to date, this is the first study to compare protocols from the literature for extracting EPS from granular filter media. Five extraction protocols were compared, and one was improved by varying the type of initial physical treatment and the shaking intensity, temperature, and time of incubation. Extracting EPS from granular filter media in triplicate by combining 2 g (wet weight) of filter media with 10 mL of extraction buffer (10 mM Tris, 10 mM EDTA, 2.5% NaCl, pH 8), vortexing for 1 min, and incubating for 4 h at 35 °C with shaking at 200 rpm yielded significantly higher EPS polysaccharide and/or protein concentrations than did protocols from the literature. This improved protocol extracted a significant fraction of the biofilm attached to sand but was less effective on biofilm attached to anthracite or granular activated carbon (GAC). A survey of 11 full-scale biofilters from the U.S. and Canada revealed that EPS polysaccharide and protein concentrations can vary over one order of magnitude, from 0.02 to 0.60 mg glucose/g total solids (TS) and 0.27 to 3.38 mg bovine serum albumin/g TS, respectively. Backwashing significantly lowered the biomass and EPS polysaccharide concentrations, but the majority of the biofilm remained attached to the filter media after backwashing, as expected. The fraction of EPS polysaccharides and proteins removed during backwashing did not differ between anthracite and GAC biofilters. The improved EPS extraction protocol can be utilized to investigate the role of EPS in biofilter performance.