Metagenomic Comparisons between Soft and Hard Feces of Plateau Pikas (Ochotona curzoniae)

SIMPLE SUMMARY: Plateau pika produces hard and soft feces with different morphology, component and microbial structure. Hard feces had more abundant Firmicutes, while soft feces had more abundant Akkermansia. The differences of microbial communities between hard and soft feces were mainly driven by core microbomes. Soft feces had a comprehensive advances in predict functional pathways compared to hard feces, these strengthened functional pathways were closely associated with metabolism of energy, vitamins, and amino acid. Our study preliminarily explored the differences in microbial structure and function between hard and soft feces, provided a foundation for future systematic explorations of the cecotrophy. ABSTRACT: The division of hard and soft feces is an effective digestion strategy in the order Lagomorpha. Although previous studies have reported that hard and soft feces differ in morphology and component, the discrepancy in the microbiome remains unclear. This study explored the microbiomes of hard and soft feces in plateau pikas by sequencing the V3 and V4 regions of 16S rDNA. We found that hard feces harbored higher Firmicutes, while soft feces harbored higher Akkermansia. Increased rare bacterial taxa were observed in hard feces compared with soft feces. Moreover, hard and soft feces displayed a greater difference in terms of core operational taxonomy units (OTUs) compared to the total OTUs. The soft feces showed enhancements in all predicted Kyoto Encyclopedia of Genes and Genomes (KEGG) functions, indicating an advancing microbial metabolism compared to hard feces. The significantly upregulated pathways in soft feces were mainly enriched in metabolism of energy and carbohydrate, glycan biosynthesis, cofactors and vitamins, and amino acids—all of which are associated with increased contents of microbial proteins, vitamins, and short-chain fatty acids. Our study reports, for the first time, the differential microbiomes between hard and soft feces of pikas and provides direction for the future studies on cecotrophy.