Faecal water was prepared and total faecal protein quantified similarly to a method previously-described41 (link), but with the addition of bacterial and mammalian protease inhibitor cocktails (G Biosciences, Uttar Pradesh, India), as well as Dithiothreitol to 1 mM final concentration (Roche, Basel, Switzerland) (to minimise enzyme oxidation42 (link)).
The BSH assay has been described previously37 (link). In brief, BSH activity was determined by measuring insoluble DCA precipitated (determined by absorbance at 600 nm (A600)) following incubation of 500 µg of faecal protein with taurodeoxycholic acid (Sigma-Aldrich, St Louis, US). Samples were compared with a standard curve of known DCA concentrations and measured in triplicate.
The BSH assay has been described previously37 (link). In brief, BSH activity was determined by measuring insoluble DCA precipitated (determined by absorbance at 600 nm (A600)) following incubation of 500 µg of faecal protein with taurodeoxycholic acid (Sigma-Aldrich, St Louis, US). Samples were compared with a standard curve of known DCA concentrations and measured in triplicate.
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