Method to Partition Between Attached and Unattached E. coli in Runoff From Agricultural Lands1

Abstract: Remediation of waters impaired by bacterial indicators is usually dictated by total maximum daily load plans, which are heavily dependent on fate and transport modeling of bacterial indicators. Nonpoint source pollution models are most frequently used to assess bacterial transport to surface waters and most models typically simulate bacterial transport as a dissolved pollutant. Previous studies have found that cells preferentially attach to sediments; however, a variety of techniques have been used to assess attachment including filtration, fractional filtration, and centrifugation. In addition, a variety of chemical and physical dispersion techniques are used to release attached and bioflocculated cells from particulates. Here we developed and validated an easy‐to‐replicate laboratory procedure for separation of unattached from attached E. coli which will also identify particle sizes to which E. coli preferentially attach. Physical and chemical dispersion techniques were evaluated and a combined hand shaker treatment for 10 min followed by dilutions in 1,000 mg/l of Tween 85 significantly increased total E. coli concentrations by 31% when compared with a control. In order to separate unattached from attached fractions, two commonly used techniques, fractional filtration, and centrifugation were combined. The filtration and centrifugation treatments did not reduce E. coli concentrations when compared with a control (p > 0.05), indicating that damage was not inflicted upon the E. coli cells during the separation procedure.