Isolation of wheat bran-colonizing and metabolizing species from the human fecal microbiota

PeerJ. 2019 Jan 25:7:e6293. doi: 10.7717/peerj.6293. eCollection 2019.

Abstract

Undigestible, insoluble food particles, such as wheat bran, are important dietary constituents that serve as a fermentation substrate for the human gut microbiota. The first step in wheat bran fermentation involves the poorly studied solubilization of fibers from the complex insoluble wheat bran structure. Attachment of bacteria has been suggested to promote the efficient hydrolysis of insoluble substrates, but the mechanisms and drivers of this microbial attachment and colonization, as well as subsequent fermentation remain to be elucidated. We have previously shown that an individually dependent subset of gut bacteria is able to colonize the wheat bran residue. Here, we isolated these bran-attached microorganisms, which can then be used to gain mechanistic insights in future pure culture experiments. Four healthy fecal donors were screened to account for inter-individual differences in gut microbiota composition. A combination of a direct plating and enrichment method resulted in the isolation of a phylogenetically diverse set of species, belonging to the Bacteroidetes, Firmicutes, Proteobacteria and Actinobacteria phyla. A comparison with 16S rRNA gene sequences that were found enriched on wheat bran particles in previous studies, however, showed that the isolates do not yet cover the entire diversity of wheat-bran colonizing species, comprising among others a broad range of Prevotella, Bacteroides and Clostridium cluster XIVa species. We, therefore, suggest several modifications to the experiment set-up to further expand the array of isolated species.

Keywords: Enrichment; Human gut microbiota; Insoluble dietary particles; Wheat bran-attached microbiota; Wheat bran-utilizing microbiota.

Grants and funding

This research was funded by the Research Foundation Flanders (FWO, Grant id=IWT130028, title=SBO BRANDING) and the Special Research Fund (BOF) Concerted Research Actions (GOA, BOF17/GOA/032) from the Flemish Government. The Phenom SEM instrument was supported by funding from Research Foundation Flanders (FWO grant G031416N to FJRM). The Hercules Foundation provided financial support in the acquisition of the scanning electron microscope JEOL JSM-7100F equipped with the cryo-transfer system Quorum PP3010T (grant no. AUGE-09-029). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.