Neutral sugar composition and gravimetric yield of plant and bacterial fractions of feces

Abstract

Separating dietary fiber from other polysaccharides in digesta and feces is necessary to understand its mechanisms of action. A gravimetric method that separates fecal plant and bacterial matter based on size and density was evaluated and modified to determine the plant and bacterial mass of lyophilized whole and blended rat and human feces. Three screen mesh combinations (150 and 75 μm, 150 and 35 μm, 35 μm) were used with rat feces. Filtration of a homogenized rat fecal slurry sequentially through 150- and 35-μm-mesh screens versus 150- and 75-μm-mesh screens decreased the gravimetric recovery of bacteria from ~35 to ~25% of fecal dry weight and increased the plant fraction weight. Neutral sugar composition, determined by gas chromatography of alditol acetates, and bacterial counts of the fractions suggested that the decreased yield of bacterial fraction represented removal of plant material and not a loss of bacteria. Rat excreta contained 29.5% (dry weight) total neutral sugar, 88% of which was recovered in the plant material. Human feces containing wheat bran, fractionated with the 150- and 35-μm-mesh screens, was 21% neutral sugar ~65% of which was in the plant fraction. The plant fractions had more xylose and arabinose and less glucose than the bacterial fractions. Processing samples in a Waring blender had no adverse effect on the rat or human fecal bacterial counts. The use of this gravimetric method in combination with the sugar analysis of the fractions provided a better measure of plant and bacteria than only gravimetric yield.