Project description:Protozoa comprise a major fraction of the microbial biomass in the rumen microbiome, of which the entodiniomorphs (order: Entodiniomorphida) and holotrichs (order: Vestibuliferida) are consistently observed to be dominant across a diverse genetic and geographical range of ruminant hosts. Despite the apparent core role that protozoal species exert, their major biological and metabolic contributions to rumen function remain largely undescribed in vivo. Here, we have leveraged (meta)genome27 centric metaproteomes from rumen fluid samples originating from both cattle and goats fed diets with varying inclusion levels of lipids and starch, to detail the specific metabolic niches that protozoa occupy in the context of their microbial co-habitants. Initial proteome estimations via total protein counts and label-free quantification highlight that entodiniomorph species Entodinium and Epidinium as well as the holotrichs Dasytricha and Isotricha comprises an extensive fraction of the total rumen metaproteome. Proteomic detection of protozoal metabolism such as hydrogenases (Dasytricha, Isotricha, Epidinium, Enoploplastron), carbohydrate-active enzymes (Epidinium, Diplodinium, Enoploplastron, Polyplastron), microbial predation (Entodinium) and volatile fatty acid production (Entodinium and Epidinium) was observed at increased levels in high methane-emitting animals. Despite certain protozoal species having well-established reputations for digesting starch, they were unexpectedly less detectable in low methane emitting- 37 animals fed high starch diets, which were instead dominated by propionate/succinate-producing bacterial populations suspected of being resistant to predation irrespective of host. Finally, we reaffirmed our abovementioned observations in geographically independent datasets, thus illuminating the substantial metabolic influence that under-explored eukaryotic populations have in the rumen, with greater implications for both digestion and methane metabolism.

Project description:Protozoa comprise a major fraction of the microbial biomass in the rumen microbiome, of which the entodiniomorphs (order: Entodiniomorphida) and holotrichs (order: Vestibuliferida) are consistently observed to be dominant across a diverse genetic and geographical range of ruminant hosts. Despite the apparent core role that protozoal species exert, their major biological and metabolic contributions to rumen function remain largely undescribed in vivo. Here, we have leveraged (meta)genome27 centric metaproteomes from rumen fluid samples originating from both cattle and goats fed diets with varying inclusion levels of lipids and starch, to detail the specific metabolic niches that protozoa occupy in the context of their microbial co-habitants. Initial proteome estimations via total protein counts and label-free quantification highlight that entodiniomorph species Entodinium and Epidinium as well as the holotrichs Dasytricha and Isotricha comprises an extensive fraction of the total rumen metaproteome. Proteomic detection of protozoal metabolism such as hydrogenases (Dasytricha, Isotricha, Epidinium, Enoploplastron), carbohydrate-active enzymes (Epidinium, Diplodinium, Enoploplastron, Polyplastron), microbial predation (Entodinium) and volatile fatty acid production (Entodinium and Epidinium) was observed at increased levels in high methane-emitting animals. Despite certain protozoal species having well-established reputations for digesting starch, they were unexpectedly less detectable in low methane emitting-animals fed high starch diets, which were instead dominated by propionate/succinate-producing bacterial populations suspected of being resistant to predation irrespective of host. Finally, we reaffirmed our abovementioned observations in geographically independent datasets, thus illuminating the substantial metabolic influence that under-explored eukaryotic populations have in the rumen, with greater implications for both digestion and methane metabolism.

Project description:Background: The regulatory effects of glucose absorption at different sites (rumen vs. small intestine) on lipid metabolism exhibit significant variation in beef nutrition. This study aimed to investigate the regulatory pathways of rumen-protected or unprotected glucose on lipid metabolism through the rumen-jejunum axis in Xinjiang Brown cattle.Results: Thirty Xinjiang Brown cattle (females) with similar initial weight (410 ± 22.4 kg) were randomly assigned to 3 treatment groups (n = 10 animals per group, biological replicates). The experimental groups were fed a basal diet (Group A; CON), a basal diet supplemented with 150 g/d of rumen-unprotected glucose (Group B), or a basal diet supplemented with 300 g/d of rumen-protected glucose (50% glucose content; Group C) for 70 days. Supplementation with both rumen-unprotected glucose and rumen-protected glucose increased chest width (P = 0.001), chest girth (P = 0.013), abdominal girth (P = 0.002), backfat thickness (P = 0.041), omental fat weight (P = 0.047), as well as serum concentrations of insulin (P < 0.001), glucagon-like peptide-1 (P < 0.001), and 5-hydroxytryptamine (P < 0.001), while decreasing the content of deoxycholic acid (P < 0.001) in the jejunum. The inclusion of rumen-unprotected glucose resulted in a trend toward higher intramuscular fat (IMF) (P = 0.064) in the longissimus thoracis, along with significant increases in C14:0 (P = 0.042), C15:0 (P = 0.014), and marbling score (P = 0.048), as well as significant reductions in drip loss (P = 0.022) and shear force (P < 0.041). These changes were accompanied by significant increases in dry matter intake (P = 0.001), ruminal concentrations of acetate (P = 0.022) and propionate (P = 0.011). The rumen-protected glucose supplementation elevated serum glucose (P < 0.001) level, while enhancing digestibility of ether extract (P = 0.027) and neutral detergent fiber (P = 0.027). Neither rumen-unprotected glucose nor rumen-protected glucose affected the α-diversity and ß-diversity of ruminal and jejunal microbiota (P > 0.05), but the differential bacterial biomarkers were either positively and negatively correlated with chest girth, abdominal girth, marbling score, backfat thickness, and deoxycholic acid (P < 0.05).Conclusion: Rumen-unprotected glucose supplementation enhances IMF deposition and meat quality attributes through microbial volatile fatty acid-driven metabolic reprogramming.

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Oral metagenome

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