Abstract

BackgroundLimited accessibility to intestinal epithelial tissue in wild animals and humans makes it challenging to study patterns of intestinal gene regulation, and hence to monitor physiological status and health in field conditions. To explore solutions to this limitation, we have used a noninvasive approach via fecal RNA-seq, for the quantification of gene expression markers in gastrointestinal cells of free-range primates and a forager human population. Thus, a combination of poly(A) mRNA enrichment and rRNA depletion methods was used in tandem with RNA-seq to quantify and compare gastrointestinal gene expression patterns in fecal samples of wild Gorilla gorilla gorilla (n = 9) and BaAka hunter-gatherers (n = 10) from The Dzanga Sangha Protected Areas, Central African Republic.ResultsAlthough only a small fraction (< 4.9%) of intestinal mRNA signals was recovered, the data was sufficient to detect significant functional differences between gorillas and humans, at the gene and pathway levels. These intestinal gene expression differences were specifically associated with metabolic and immune functions. Additionally, non-host RNA-seq reads were used to gain preliminary insights on the subjects’ dietary habits, intestinal microbiomes, and infection prevalence, via identification of fungi, nematode, arthropod and plant RNA.ConclusionsOverall, the results suggest that fecal RNA-seq, targeting gastrointestinal epithelial cells can be used to evaluate primate intestinal physiology and gut gene regulation, in samples obtained in challenging conditions in situ. The approach used herein may be useful to obtain information on primate intestinal health, while revealing preliminary insights into foraging ecology, microbiome, and diet.

Highlights

  • Limited accessibility to intestinal epithelial tissue in wild animals and humans makes it challenging to study patterns of intestinal gene regulation, and to monitor physiological status and health in field conditions

  • After quality control and alignment of pair-end reads to the latest genome drafts for G. g. gorilla and H. sapiens (GRCh38/hg38), the percentage of reads aligning to the respective host genomes ranged from 0. 44 to 1.41% for samples undergoing poly(A) Messenger RNA (mRNA) enrichment plus Ribosomal RNA (rRNA) depletion

  • Here, we attempted to maximize functional genomic information from the intestinal tract of wild lowland gorillas and the BaAka foragers from the Dzanga Sangha Protected Areas in the Central African Republic

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Summary

Introduction

Limited accessibility to intestinal epithelial tissue in wild animals and humans makes it challenging to study patterns of intestinal gene regulation, and to monitor physiological status and health in field conditions. To explore solutions to this limitation, we have used a noninvasive approach via fecal RNA-seq, for the quantification of gene expression markers in gastrointestinal cells of free-range primates and a forager human population. Difficulties in obtaining nucleic acids from representative samples make it necessary to implement non-invasive approaches to explore genetic traits in wild animals and humans [1, 2]. Apart from difficulties in obtaining representative tissue samples, significant challenges associated with these analyses include preservation and integrity of samples and nucleic acids in field conditions, contamination with other biological materials in feces (e.g. microbes, dietary materials) and inability to detect. Other techniques, such as droplet digital PCR, have been useful in detecting transcriptional markers of inflammation from human fecal samples [20]

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