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About 200 mg of feces were diluted in 1 ml of distilled water, homogenized and centrifuged at 1000 g for 5 min. One hundred microliters of fecal supernatants were added to 100 µl of buffer (50 mM Tris-HCl pH 8, 1 mM CaCl2) containing 100 µM specific serine protease substrate (suc-phe-ala-ala-phe-pNA) (Bachem, Switzerland; Cat. No. 4013859) and incubated at 37 °C for 6 h. Substrate cleavage was measured at 400 nm with NUNC 96-well plates (ThermoFisher Scientific, Waltham, MS, USA) and normalized to the total feces weight.
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